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Mario LUPPI

Professore Ordinario
Dipartimento di Scienze Mediche e Chirurgiche Materno-Infantili e dell'Adulto

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Pubblicazioni

- "Metodo per la diagnosi e/o il monitoraggio della mucormicosi" - "Method for the diagnosis of and/or monitoring mucormycosis" [Brevetto]
Luppi, Mario; Potenza, Leonardo; Barozzi, Patrizia; Vallerini, Daniela; Forghieri, Fabio
abstract

A method is described for the diagnosis and/or monitoring of active or previous infection by Mucor which consists in the identification of Mucorales-specific T cells in samples from biological fluids taken from the patient and put into contact with a Mucor antigen. These specific immune responses can be detected by the execution of immunoenzymatic assays (ELISPOT, Quantiferon) or of immunocytofluorimetric assays [Cytokine Secretion Assay (CSA), Intracellular Cytokine Staining (ICS)] in vitro. In greater detail, the method in question provides for checking for the presence of specific IFN-γ producing T cells, of specific IL-10 producing T cells and/or specific IL- 4 producing T cells.

- Metodo per la diagnosi e/o il monitoraggio dell’ aspergillosi invasiva [Brevetto]
Torelli, Giuseppe; Luppi, Mario; Barozzi, Patrizia; Potenza, Leonardo
abstract

La presente invenzione si riferisce ad un metodo per la diagnosi e/o ilmonitoraggio dell’infezione attiva o pregressa da Aspergillus fumigatus checomprende l’esecuzione di un saggio immunoenzimatico in vitro (ELISPOT) incui il campione da fluidi biologici prelevato dal paziente è messo in contatto conun antigene di Aspergillus fumigatus.

2023 - 10-day decitabine versus 3 + 7 chemotherapy followed by allografting in older patients with acute myeloid leukaemia: an open-label, randomised, controlled, phase 3 trial [Articolo su rivista]
Lubbert, M.; Wijermans, P. W.; Kicinski, M.; Chantepie, S.; Van der Velden, W. J. F. M.; Noppeney, R.; Griskevicius, L.; Neubauer, A.; Crysandt, M.; Vrhovac, R.; Luppi, M.; Fuhrmann, S.; Audisio, E.; Candoni, A.; Legrand, O.; Foa, R.; Gaidano, G.; van Lammeren-Venema, D.; Posthuma, E. F. M.; Hoogendoorn, M.; Giraut, A.; Stevens-Kroef, M.; Jansen, J. H.; de Graaf, A. O.; Efficace, F.; Ammatuna, E.; Vilque, J. -P.; Wasch, R.; Becker, H.; Blijlevens, N.; Duhrsen, U.; Baron, F.; Suciu, S.; Amadori, S.; Venditti, A.; Huls, G.; Finke, J.; Schaap, N. P. M.; Zucenka, A.; Metzelder, S.; Jost, E.; Peric, Z.; Forghieri, F.; Allione, B.; Martelli, M.; Iori, A. P.; Wittnebel, S.; Mengarelli, A.; Imovilli, A.; Olivieri, A.; De Prijck, B. J. M.; van der Poel, M. W. M.; Junghanss, C.; Salih, H. R.; Tafuri, A.; Guimaraes, J. E.; Musso, M.; De Fabritiis, P.; Chevallier, P.; Selleslag, D. L.; Cascavilla, N.; Berneman, Z.; Jaspers, A.; Zuffa, E.; Vanstraelen, G.; Visani, G.; Cuijpers, M. L. H.; De Becker, A.; Mianulli, A. M.; Hackanson, B.; Mihaylov, G. G.; Martinelli, G.; Paolini, S.; Zinzani, P. L.; Henkes, M.; Al-Ali, H. K.; La Rosee, P.; Chierichini, A.; Cudillo, L.; Specchia, G.; Simec, N. G.; Capalbo, S. F.; Spinosa, G.; Molica, S.; de Jonge-Peeters, S. D.
abstract

Background: Many older patients with acute myeloid leukaemia die or cannot undergo allogeneic haematopoietic stem-cell transplantation (HSCT) due to toxicity caused by intensive chemotherapy. We hypothesised that replacing intensive chemotherapy with decitabine monotherapy could improve outcomes. Methods: This open-label, randomised, controlled, phase 3 trial was conducted at 54 hospitals in nine European countries. Patients aged 60 years and older who were newly diagnosed with acute myeloid leukaemia and had not yet been treated were enrolled if they had an Eastern Cooperative Oncology Group performance status of 2 or less and were eligible for intensive chemotherapy. Patients were randomly assigned (1:1) to receive decitabine or standard chemotherapy (known as 3 + 7). For the decitabine group, decitabine (20 mg/m2) was administered for the first 10 days in the first 28-day cycle, followed by 28-day cycles consisting of 5 days or 10 days of decitabine. For the 3 + 7 group, daunorubicin (60 mg/m2) was administered over the first 3 days and cytarabine (200 mg/m2) over the first 7 days, followed by 1–3 additional chemotherapy cycles. Allogeneic HSCT was strongly encouraged. Overall survival in the intention-to-treat population was the primary endpoint. Safety was assessed in all patients who received the allocated treatment. This trial is registered at ClinicalTrials.gov, NCT02172872, and is closed to new participants. Findings: Between Dec 1, 2014, and Aug 20, 2019, 606 patients were randomly assigned to the decitabine (n=303) or 3 + 7 (n=303) group. Following an interim analysis which showed futility, the IDMC recommended on May 22, 2019, that the study continued as planned considering the risks and benefits for the patients participating in the study. The cutoff date for the final analysis presented here was June 30, 2021. At a median follow-up of 4·0 years (IQR 2·9–4·8), 4-year overall survival was 26% (95% CI 21–32) in the decitabine group versus 30% (24–35) in the 3 + 7 group (hazard ratio for death 1·04 [95% CI 0·86–1·26]; p=0·68). Rates of on-protocol allogeneic HSCT were similar between groups (122 [40%] of 303 patients for decitabine and 118 [39%] of 303 patients for 3+7). Rates of grade 3–5 adverse events were 254 (84%) of 302 patients in the decitabine group and 279 (94%) of 298 patients in the 3 + 7 group. The rates of grade 3–5 infections (41% [125 of 302] vs 53% [158 of 298]), oral mucositis (2% [seven of 302] vs 10% [31 of 298]) and diarrhoea (1% [three of 302] vs 8% [24 of 298]) were lower in the decitabine group than in the 3 + 7 group. Treatment-related deaths were reported for 12% (35 of 302) of patients in the decitabine group and 14% (41 of 298) in the 3 + 7 group. Interpretation: 10-day decitabine did not improve overall survival but showed a better safety profile compared with 3 + 7 chemotherapy in older patients with acute myeloid leukaemia eligible for intensive chemotherapy. Decitabine could be considered a better-tolerated and sufficiently efficacious alternative to 3 + 7 induction in fit older patients with acute myeloid leukaemia without favourable genetics. Funding: Janssen Pharmaceuticals.

2023 - Abstracts from the 55th European Society of Human Genetics (ESHG) Conference: Hybrid Posters [Abstract in Rivista]
Tenedini, Elena; Piana, Simonetta; Toss, Angela; Marino, Marco; Barbieri, Elena; Artuso, Lucia; Venturelli, Marta; Gasparini, Elisa; Dario Mandato, Vincenzo; Marchi, Isabella; Castellano, Sara; Luppi, Mario; Trenti, Tommaso; Cortesi, Laura; Tagliafico, Enrico
abstract

2023 - Busulfan or Treosulfan Conditioning Platform for Allogeneic Stem Cell Transplantation in Patients Aged >60 Y With Acute Myeloid Leukemia/Myelodysplastic Syndrome: A Subanalysis of the GITMO AlloEld Study [Articolo su rivista]
Malagola, Michele; Polverelli, Nicola; Martino, Massimo; Patriarca, Francesca; Bruno, Benedetto; Giaccone, Luisa; Grillo, Giovanni; Bramanti, Stefania; Bernasconi, Paolo; De Gobbi, Marco; Natale, Annalisa; Terruzzi, Elisabetta; Olivieri, Attilio; Chiusolo, Patrizia; Carella, Angelo Michele; Casini, Marco; Maffini, Enrico; Nozzoli, Chiara; Mazza, Patrizio; Bassi, Simona; Onida, Francesco; Vacca, Adriana; Falcioni, Sadia; Luppi, Mario; Iori, Anna Paola; Pavone, Vincenzo; Skert, Cristina; Carluccio, Paola; Borghero, Carlo; Proia, Anna; Selleri, Carmine; Rubini, Vicky; Sacchi, Nicoletta; Oldani, Elena; Bonifazi, Francesca; Ciceri, Fabio; Russo, Domenico
abstract

Background.The conditioning regimens with different alkylators at different doses can influence the outcome of allogeneic stem cell transplantation (SCT), but conclusive data are missing. Methods.With the aim to analyze real-life allogeneic SCTs performed in Italy between 2006 and 2017 in elderly patients (aged >60 y) with acute myeloid leukemia or myelodysplastic syndrome, we collected 780 first transplants data. For analysis purposes, patients were grouped according to the type of alkylator included in the conditioning (busulfan [BU]-based; n = 618; 79%; treosulfan [TREO]-based; n=162; 21%). Results.No significant differences were observed in nonrelapse mortality, cumulative incidence of relapse, and overall survival, although in the TREO-based group, we observed a greater proportion of elderly patients (P < 0.001); more active diseases at the time of SCT (P < 0.001); a higher prevalence of patients with either hematopoietic cell transplantation-comorbidity index >= 3 (P < 0.001) or a good Karnofsky performance status (P = 0.025); increased use of peripheral blood stem cells as graft sources (P < 0.001); and greater use of reduced intensity conditioning regimens (P = 0.013) and of haploidentical donors (P < 0.001). Moreover, the 2-y cumulative incidence of relapse with myeloablative doses of BU was significantly lower than that registered with reduced intensity conditioning (21% versus 31%; P = 0.0003). This was not observed in the TREO-based group. Conclusions.Despite a higher number of risk factors in the TREO group, no significant differences were observed in nonrelapse mortality, cumulative incidence of relapse, and overall survival according to the type of alkylator, suggesting that TREO has no advantage over BU in terms of efficacy and toxicity in acute myeloid leukemia and myelodysplastic syndrome.

2023 - COVID-19 omicron variant outbreak in a hematopoietic stem cell transplant unit [Articolo su rivista]
Gilioli, A.; Bresciani, P.; Franceschini, E.; Messerotti, A.; Pioli, V.; Colasante, C.; Bettelli, F.; Giusti, D.; Forghieri, F.; Morselli, M.; Colaci, E.; Potenza, L.; Gennari, W.; Pecorari, M.; Marasca, R.; Candoni, A.; Mussini, C.; Trenti, T.; Comoli, P.; Luppi, M.; Cuoghi, A.
abstract

Recommendations and guidelines for management of SARS-COV-2 infection in hematologic patients were developed in the very difficult context of dealing with novel viral variants from one pandemic wave to another, with different susceptibility to available drugs and vaccines. Moreover, the largest SARS-COV-2 case series in patients treated for hematologic malignancies, including stem cell transplant recipients, was published before the Omicron surge, and refers mainly to Alpha and Delta viral variants. These infections had very high mortality, in a period when antivirals and monoclonal antibodies were mostly unavailable. Here, we report for the first time a SARS-COV-2 Omicron variant outbreak inside a Bone Marrow Transplant (BMT) Unit, describing the characteristics, clinical course, and infection outcomes shortly before and shortly after myeloablative transplantation. We detail how infections were treated off-label and managed inside the BMT ward, to guarantee the best possible outcomes while avoiding risks for non-infected inpatients. The positive outcomes observed suggest that it may not be absolutely necessary to obtain SARS-CoV-2 PCR negativity before BMT in hematologic patients after treated infection, in cases with long-term PCR positivity and high-risk hematologic disease.

2023 - Characteristics and clinical behavior of acute myeloid leukemia harboring rare non-A/B/D nucleophosmin (NPM1) gene mutation subtypes: a single-center experience and review of the literature [Articolo su rivista]
Mutti, M.; Cordella, S.; Parisotto, A.; Bettelli, F.; Morselli, M.; Cuoghi, A.; Bresciani, P.; Messerotti, A.; Gilioli, A.; Pioli, V.; Giusti, D.; Colaci, E.; Cassanelli, L.; Paolini, A.; Martinelli, S.; Maffei, R.; Riva, G.; Nasillo, V.; Sarti, M.; Trenti, T.; Comoli, P.; Tagliafico, E.; Manfredini, R.; Eccher, A.; Lagreca, I.; Barozzi, P.; Potenza, L.; Marasca, R.; Candoni, A.; Luppi, M.; Forghieri, F.
abstract

2023 - Chromosome 9p Duplication Promotes T-Cell Exhaustion and Enhances Stem Cell Clonogenic Potential in JAK2-Mutant Myeloproliferative Neoplasms [Abstract in Rivista]
Norfo, Ruggiero; Carretta, Chiara; Parenti, Sandra; Badii, Filippo; Bertesi, Matteo; Rontauroli, Sebastiano; Tavernari, Lara; Genovese, Elena; Sperduti, Samantha; Enzo, Elena; Mirabile, Margherita; Pedrazzi, Francesca; Pessina, Chiara; Colugnat, Ilaria; Mora, Barbara; Maccaferri, Monica; Tenedini, Elena; Martinelli, Silvia; Bianchi, Elisa; Casarini, Livio; Potenza, Leonardo; Luppi, Mario; Tagliafico, Enrico; Guglielmelli, Paola; Simoni, Manuela; Passamonti, Francesco; Vannucchi, Alessandro Maria; Manfredini, Rossella
abstract

2023 - Clinically relevant low-frequency Next Generation Sequencing variants in hereditary cancer patients: an operational multi-step algorithm for laboratory managing [Articolo su rivista]
Tenedini, E.; Bonamici, L.; Marino, M.; Artuso, L.; Luppi, M.; Trenti, T.; Tagliafico, E.
abstract

2023 - Do GWAS-Identified Risk Variants for Chronic Lymphocytic Leukemia Influence Overall Patient Survival and Disease Progression? [Articolo su rivista]
Cabrera-Serrano, Antonio José; Sánchez-Maldonado, José Manuel; Ter Horst, Rob; Macauda, Angelica; García-Martín, Paloma; Benavente, Yolanda; Landi, Stefano; Clay-Gilmour, Alyssa; Niazi, Yasmeen; Espinet, Blanca; Rodríguez-Sevilla, Juan José; Pérez, Eva María; Maffei, Rossana; Blanco, Gonzalo; Giaccherini, Matteo; Cerhan, James R; Marasca, Roberto; López-Nevot, Miguel Ángel; Chen-Liang, Tzu; Thomsen, Hauke; Gámez, Irene; Campa, Daniele; Moreno, Víctor; de Sanjosé, Silvia; Marcos-Gragera, Rafael; García-Álvarez, María; Dierssen-Sotos, Trinidad; Jerez, Andrés; Butrym, Aleksandra; Norman, Aaron D; Luppi, Mario; Slager, Susan L; Hemminki, Kari; Li, Yang; Berndt, Sonja I; Casabonne, Delphine; Alcoceba, Miguel; Puiggros, Anna; Netea, Mihai G; Försti, Asta; Canzian, Federico; Sainz, Juan
abstract

Chronic lymphocytic leukemia (CLL) is the most common leukemia among adults worldwide. Although genome-wide association studies (GWAS) have uncovered the germline genetic component underlying CLL susceptibility, the potential use of GWAS-identified risk variants to predict disease progression and patient survival remains unexplored. Here, we evaluated whether 41 GWAS-identified risk variants for CLL could influence overall survival (OS) and disease progression, defined as time to first treatment (TTFT) in a cohort of 1039 CLL cases ascertained through the CRuCIAL consortium. Although this is the largest study assessing the effect of GWAS-identified susceptibility variants for CLL on OS, we only found a weak association of ten single nucleotide polymorphisms (SNPs) with OS (p < 0.05) that did not remain significant after correction for multiple testing. In line with these results, polygenic risk scores (PRSs) built with these SNPs in the CRuCIAL cohort showed a modest association with OS and a low capacity to predict patient survival, with an area under the receiver operating characteristic curve (AUROC) of 0.57. Similarly, seven SNPs were associated with TTFT (p < 0.05); however, these did not reach the multiple testing significance threshold, and the meta-analysis with previous published data did not confirm any of the associations. As expected, PRSs built with these SNPs showed reduced accuracy in prediction of disease progression (AUROC = 0.62). These results suggest that susceptibility variants for CLL do not impact overall survival and disease progression in CLL patients.

2023 - Early palliative care for solid and blood cancer patients and caregivers: Quantitative and qualitative results of a long-term experience as a case of value-based medicine [Articolo su rivista]
Bigi, Sarah; Borelli, Eleonora; Potenza, Leonardo; Gilioli, Fabio; Artioli, Fabrizio; Porzio, Giampiero; Luppi, Mario; Bandieri, Elena
abstract

Introduction: Cancer patients and their caregivers have substantial unmet needs, that negatively impact the clinical outcome and quality of life. However, interventions aimed to address such needs are still suboptimal, failing to answer the recent healthcare call for the adoption of value-based models of care. In the case of incurable oncologic and hematologic cancers, a value-based model of care should plan advanced care on patients' needs and include the quality of death as an outcome. The integration of early palliative care into standard oncologic care for patients with advanced cancers represents a recent innovative model of assistance whose benefits for patients and caregivers are now widely recognized. The key elements underlying the reasons behind these benefits are the multidisciplinary collaboration (teamwork), an honest and empathetic communication between the early palliative care team, the patient, and the caregiver (rapport building), and the ability to detect changes in the physical/psychosocial wellbeing of the patient, along the whole disease trajectory (constant monitoring). Methods: This community case study documents the quantitative and qualitative results of a long term clinical and research experience in delivering early palliative care service to address both solid and blood cancer patients' and their primary caregivers' needs. Results: Data showed decreased use of chemotherapy, blood transfusions and referral to intensive care units near the end of life; increased life expectancy; improved symptom burden and mood; increased frequency of goals-of-care and advanced care planning conversations. Hope perception among bereaved caregivers was associated with resilience and realistic expectations raising from honest communication with the early palliative care team and appreciation toward the model. Patients and caregivers perceived the possibility of a good death as realistic and not as an unlikely event as it was for patients and caregivers on standard oncologic care only. Gratitude expressions toward the model and the team were frequently identified in their reports and positively associated with communication and spirituality. Conclusions: These findings are discussed in the context of an updated literature review regarding value-based care and suggest that early palliative care integrated into standard oncology care may be considered as an effective model of value-based care.

2023 - Early palliative care versus usual haematological care in multiple myeloma: retrospective cohort study [Articolo su rivista]
Giusti, D.; Colaci, E.; Pioli, V.; Banchelli, F.; Maccaferri, M.; Leonardi, G.; Marasca, R.; Morselli, M.; Forghieri, F.; Bettelli, F.; Cuoghi, A.; Bresciani, P.; Messerotti, A.; Gilioli, A.; Candoni, A.; Cassanelli, L.; Sbadili, E.; Bassoli, I.; Longo, G.; Gilioli, F.; Borelli, E.; Bigi, S.; D'Amico, R.; Porro, C. A.; Odejide, O.; Zimmermann, C.; Efficace, F.; Bruera, E.; Luppi, M.; Bandieri, E.; Potenza, L.
abstract

Objectives Although early palliative care (EPC) is beneficial in acute myeloid leukaemia, little is known about EPC value in multiple myeloma (MM). We compared quality indicators for palliative and end of life (EOL) care in patients with MM receiving EPC with those of patients who received usual haematological care (UHC).Methods This observational, retrospective study was based on 290 consecutive patients with MM. The following indicators were abstracted: providing psychological support, assessing/managing pain, discussing goals of care, promoting advance care plan, accessing home care services; no anti MM treatment within 14 and 30 days and hospice length of stay >7 days before death; no cardiopulmonary resuscitation, no intubation, <2 hospitalisations and emergency department visits within 30 days before death. Comparisons were performed using unadjusted and confounder adjusted regression models.Results 55 patients received EPC and 231 UHC. Compared with UHC patients, EPC patients had a significantly higher number of quality indicators of care (mean 2.62 +/- 1.25 vs 1.12 +/- 0.95; p<0.0001)); a significant reduction of pain intensity over time (p<0.01) and a trend towards reduced aggressiveness at EOL, with the same survival (5.3 vs 5.46 years; p=0.74)).Conclusions Our data support the value of integrating EPC into MM routine practice and lay the groundwork for future prospective comparative studies.

2023 - Effects of the BTN162b2 mRNA COVID-19 vaccine in humoral and cellular immunity in patients with chronic lymphocytic leukemia [Articolo su rivista]
Fiorcari, S.; Atene, C. G.; Maffei, R.; Mesini, N.; Debbia, G.; Colasante, C.; Pozzi, S.; Barbieri, E.; Maccaferri, M.; Leonardi, G.; Potenza, L.; Luppi, M.; Marasca, R.
abstract

Chronic lymphocytic leukemia (CLL), the most common leukemia in the western countries, is characterized by immunosuppression due to disease itself and cytotoxic treatments. Since the beginning of COVID-19 pandemic, patients with CLL appear to be a vulnerable population. In addition, phase III mRNA vaccine trials did not provide information about the efficacy in immunocomprised population. In CLL, the antibody-mediated response to SARS-CoV-2 vaccine is impaired. The goal of this study was to evaluate the effects of SARS-CoV-2 vaccination on humoral immune response and on cellular immunity in CLL patients. Humoral immune response to BNT162b2 messenger RNA COVID-19 vaccine was evaluated in 44 CLL patients comprising 20 treatment-naïve, 14 under treatment with ibrutinib and 10 in follow-up after completion of therapy. A positive serological response to SARS-CoV-2 vaccination with IgG titers higher than 13 UA/ml was detected in 54.6% of CLL patients with a higher response in patients who obtained remission after treatment. Reduced antibody response was detected in patients under ibrutinib treatment. T-cell response to overlapping pool of peptides representing the spike region was assessed in paired CLL samples collected before and after 1 month from the second dose of COVID-19 vaccine in treatment-naïve and ibrutinib-treated CLL patients using cytokine secretion assay. Both CD3+ CD4+ and CD3+ CD8+ T cells are able to mount a cellular response to spike peptides with secretion of IFNγ and TNFα before and after vaccination in both treatment naïve and ibrutinib-treated patients and this cellular immune response is independent by COVID-19 vaccination. Collectively, T cell response to spike peptides appeared more blunted in CLL patients under treatment with ibrutinib compared to untreated ones. Our study supports the need for optimization of vaccination strategy to achieve an adequate immune response keeping strict preventive measures by CLL patients against COVID-19.

2023 - From gating to computational flow cytometry: Exploiting artificial intelligence for MRD diagnostics [Articolo su rivista]
Riva, Giovanni; Luppi, Mario; Tagliafico, Enrico
abstract

The era of AI-based methods to improve flow cytometry diagnostics in haematology is now at the beginning. The study by Nguyen and colleagues explored an emerging machine learning approach to assess phenotypic MRD in chronic lymphocytic leukaemia patients, showing that such AI-driven computational analysis may represent a robust and feasible tool for advanced diagnostics of haematological malignancies.Commentary on: Nguyen et al. Computational flow cytometry provides accurate assessment of measurable residual disease in chronic lymphocytic leukaemia. Br J Haematol 2023 (Online ahead of print). doi:

2023 - Improved efficacy of quizartinib in combination therapy with PI3K inhibition in primary FLT3-ITD AML cells [Articolo su rivista]
Darici, Salihanur; Jørgensen, Heather G; Huang, Xu; Serafin, Valentina; Antolini, Ludovica; Barozzi, Patrizia; Luppi, Mario; Forghieri, Fabio; Marmiroli, Sandra; Zavatti, Manuela
abstract

: Acute myeloid leukemia is a heterogeneous hematopoietic malignancy, characterized by uncontrolled clonal proliferation of abnormal myeloid progenitor cells, with poor outcomes. The internal tandem duplication (ITD) mutation of the Fms-like receptor tyrosine kinase 3 (FLT3) (FLT3-ITD) represents the most common genetic alteration in AML, detected in approximately 30% of AML patients, and is associated with high leukemic burden and poor prognosis. Therefore, this kinase has been regarded as an attractive druggable target for the treatment of FLT3-ITD AML, and selective small molecule inhibitors, such as quizartinib, have been identified and trialled. However, clinical outcomes have been disappointing so far due to poor remission rates, also because of acquired resistance. A strategy to overcome resistance is to combine FLT3 inhibitors with other targeted therapies. In this study, we investigated the preclinical efficacy of the combination of quizartinib with the pan PI3K inhibitor BAY-806946 in FLT3-ITD cell lines and primary cells from AML patients. We show here that BAY-806946 enhanced quizartinib cytotoxicity and, most importantly, that this combination increases the ability of quizartinib to kill CD34+ CD38-leukemia stem cells, whilst sparing normal hematopoietic stem cells. Because constitutively active FLT3 receptor tyrosine kinase is known to boost aberrant PI3K signaling, the increased sensitivity of primary cells to the above combination can be the mechanistic results of the disruption of signaling by vertical inhibition.

2023 - Inhibition of ERK1/2 signaling prevents bone marrow fibrosis by reducing osteopontin plasma levels in a myelofibrosis mouse model [Articolo su rivista]
Bianchi, Elisa; Rontauroli, Sebastiano; Tavernari, Lara; Mirabile, Margherita; Pedrazzi, Francesca; Genovese, Elena; Sartini, Stefano; Dall'Ora, Massimiliano; Grisendi, Giulia; Fabbiani, Luca; Maccaferri, Monica; Carretta, Chiara; Parenti, Sandra; Fantini, Sebastian; Bartalucci, Niccolò; Calabresi, Laura; Balliu, Manjola; Guglielmelli, Paola; Potenza, Leonardo; Tagliafico, Enrico; Losi, Lorena; Dominici, Massimo; Luppi, Mario; Vannucchi, Alessandro Maria; Manfredini, Rossella
abstract

Clonal myeloproliferation and development of bone marrow (BM) fibrosis are the major pathogenetic events in myelofibrosis (MF). The identification of novel antifibrotic strategies is of utmost importance since the effectiveness of current therapies in reverting BM fibrosis is debated. We previously demonstrated that osteopontin (OPN) has a profibrotic role in MF by promoting mesenchymal stromal cells proliferation and collagen production. Moreover, increased plasma OPN correlated with higher BM fibrosis grade and inferior overall survival in MF patients. To understand whether OPN is a druggable target in MF, we assessed putative inhibitors of OPN expression in vitro and identified ERK1/2 as a major regulator of OPN production. Increased OPN plasma levels were associated with BM fibrosis development in the Romiplostim-induced MF mouse model. Moreover, ERK1/2 inhibition led to a remarkable reduction of OPN production and BM fibrosis in Romiplostim-treated mice. Strikingly, the antifibrotic effect of ERK1/2 inhibition can be mainly ascribed to the reduced OPN production since it could be recapitulated through the administration of anti-OPN neutralizing antibody. Our results demonstrate that OPN is a novel druggable target in MF and pave the way to antifibrotic therapies based on the inhibition of ERK1/2-driven OPN production or the neutralization of OPN activity.

2023 - Midostaurin plus daunorubicin or idarubicin for young and older adults with FLT3-mutated AML: a phase 3b trial [Articolo su rivista]
Sierra, J.; Montesinos, P.; Thomas, X.; Griskevicius, L.; Cluzeau, T.; Caillot, D.; Legrand, O.; Minotti, C.; Luppi, M.; Farkas, F.; Bengoudifa, B. R.; Gilotti, G.; Hodzic, S.; Rambaldi, A.; Venditti, A.
abstract

The pivotal RATIFY study demonstrated midostaurin (50 mg twice daily) with standard chemotherapy significantly reduced mortality in adult patients (<60 years) with newly diagnosed (ND) FLT3(mut) acute myeloid leukemia (AML). Considering that AML often present in older patients who show poor response to chemotherapy, this open-label, multicenter phase 3b trial was designed to further assess safety and efficacy of midostaurin plus chemotherapy in induction, consolidation, and maintenance monotherapy in young (<= 60 years) and older (>60 years) patients with FLT3(mut) ND-AML. Compared with RATIFY, this study extended midostaurin treatment from 14 days to 21 days, substituted anthracyclines (idarubicin or daunorubicin), and introduced variation in standard combination chemotherapy dosing ("7+3" or "5+2" in more fragile patients). Total 301 patients (47.2% >60 years and 82.7% with FLT3-ITDmut) of median age 59 years entered induction phase. Overall, 295 patients (98.0%) had at least 1 adverse event (AE), including 254 patients (84.4%) with grade >= 3 AE. The grade >= 3 serious AEs occurred in 134 patients. No difference was seen in AE frequency between age groups, but grade >= 3AE frequency was higher in older patients. Overall, complete remission (CR) rate including incomplete hematologic recovery (CR + CRi) (80.7% [95% confidence interval, 75.74-84.98]) was comparable between age groups (<= 60 years [83.5%]; >60 to <= 70 years [82.5%]; in patients >70 years [64.1%]) and the type of anthracycline used in induction. CR + CRi rate was lower in males (76.4%) than females (84.4%). Overall, the safety and efficacy of midostaurin remains consistent with previous findings, regardless of age, sex, or induction regimen. The trial is registered at www.clinicaltrials.gov as #NCT03379727.

2023 - PEG-Asparaginase Single-Agent Rescue in an Advanced Case of Monomorphic Epitheliotropic Intestinal T Cell Lymphoma [Articolo su rivista]
Barbieri, E.; Pozzi, S.; Gelmini, R.; Roncati, L.; Maccaferri, M.; Potenza, L.; Marasca, R.; Luppi, M.; Leonardi, G.
abstract

Purpose: MEITL is a very rare and highly aggressive peripheral T cell lymphoma with poor prognosis and for which there is no standard treatment. Treatment options for patients patients with relapsed/refractory disease are scarce and the choice of an appropriate rescue still represents an unmet need. Methods: Here, we report the case of a 65-year-old woman affected by MEITL, progressing after initial treatment with an anthracycline-based chemotherapy and surgery, who received single-agent PEG-asparaginase salvage therapy at our institution. Results: PEG-asparaginase single-agent rescue proved to be rapidly effective in controlling the disease and its associated paraneoplastic features. Nevertheless, toxicity was high and the patient died due to a treatment-related complication. Conclusion: The case we described brings new evidences on the effectiveness of PEG-asparaginase therapy in MEITL patients. Whether PEG-asparaginase should be included in the treatment course of MEITL patients could be the subject of future studies.

2023 - Perceptions of Death Among Patients with Advanced Cancer Receiving Early Palliative Care and Their Caregivers: Results from a Mixed-Method Analysis [Articolo su rivista]
Bigi, S.; Ganfi, V.; Borelli, E.; Potenza, L.; Artioli, F.; Eliardo, S.; Mucciarini, C.; Cottafavi, L.; Ferrari, U.; Lombardo, L.; Cagossi, K.; Pietramaggiori, A.; Fantuzzi, V.; Bernardini, I.; Cruciani, M.; Cacciari, C.; Odejide, O.; Adolfo Porro, C.; Zimmermann, C.; Efficace, F.; Bruera, E.; Luppi, M.; Bandieri, E.
abstract

BACKGROUND: Oncologists are often concerned that talking about death with patients may hinder their relationship. However, the views of death held by patients have not been thoroughly investigated. This study aimed to describe the perception of death among patients with advanced cancer receiving early palliative care (EPC) and their caregivers. MATERIAL AND METHODS: Qualitative and quantitative analyses were performed on 2 databases: (a) transcripts of open-ended questionnaires administered to 130 cancer patients receiving EPC with a mean age of 68.4 years and to 115 primary caregivers of patients on EPC with a mean age of 56.8; (b) texts collected from an Italian forum, containing instances of web-mediated interactions between patients and their caregivers. RESULTS: Quantitative analysis shows that: (a) patients and caregivers are not afraid of speaking about death; (b) patients and caregivers on EPC use the word "death" significantly more than patients on standard oncology care (SOC) and their caregivers (P < .0001). For both participants on EPC and SOC, the adjectives and verbs associated with the word "death" have positive connotations; however, these associations are significantly more frequent for participants on EPC (verbs, Ps < .0001; adjectives, Ps < .003). Qualitative analysis reveals that these positive connotations refer to an actual, positive experience of the end of life in the EPC group and a wish or a negated event in the SOC group. CONCLUSIONS: EPC interventions, along with proper physician-patient communication, may be associated with an increased acceptance of death in patients with advanced cancer and their caregivers.

2023 - Persistent SARS-CoV-2 infection with multiple clinical relapses in two patients with follicular lymphoma treated with bendamustine and obinutuzumab or rituximab [Articolo su rivista]
Franceschini, Erica; Pellegrino, Mariachiara; Todisco, Vera; Dolci, Giovanni; Bettelli, Francesca; Meschiari, Marianna; Bedini, Andrea; Fregni-Serpini, Giulia; Grottola, Antonella; Guaraldi, Giovanni; Pecorari, Monica; Sarti, Mario; Luppi, Mario; Perno, Carlo Federico; Mussini, Cristina
abstract

Purpose: People with hematologic malignancies have a significantly higher risk of developing severe and protracted forms of SARS-CoV-2 infection compared to immunocompetent patients, regardless of vaccination status. Results: We describe two cases of prolonged SARS-CoV-2 infection with multiple relapses of COVID-19 pneumonia in patients with follicular lymphoma treated with bendamustine and obinutuzumab or rituximab. The aim is to highlight the complexity of SARS-CoV-2 infection in this fragile group of patients and the necessity of evidence-based strategies to treat them properly. Conclusions: Patients with hematological malignancies treated with bendamustine and anti-CD20 antibodies had a significant risk of prolonged and relapsing course of COVID-19. Specific preventive and therapeutic strategies should be developed for this group of patients.

2023 - Preclinical Validation of an Advanced Therapy Medicinal Product Based on Cytotoxic T Lymphocytes Specific for Mutated Nucleophosmin (NPM1mut) for the Treatment of NPM1mut-Acute Myeloid Leukemia [Articolo su rivista]
De Cicco, Marica; Lagreca, Ivana; Basso, Sabrina; Barozzi, Patrizia; Muscianisi, Stella; Bianco, Alba; Riva, Giovanni; Di Vincenzo, Sara; Pulvirenti, Chiara; Sapuppo, Davide; Siciliano, Mariangela; Rosti, Vittorio; Candoni, Anna; Zecca, Marco; Forghieri, Fabio; Luppi, Mario; Comoli, Patrizia
abstract

2023 - Prognostic Relevance of Multi-Antigenic Myeloma-Specific T-Cell Assay in Patients with Monoclonal Gammopathies [Articolo su rivista]
Lagreca, Ivana; Nasillo, Vincenzo; Barozzi, Patrizia; Castelli, Ilaria; Basso, Sabrina; Castellano, Sara; Paolini, Ambra; Maccaferri, Monica; Colaci, Elisabetta; Vallerini, Daniela; Natali, Patrizia; Debbia, Daria; Pirotti, Tommaso; Ottomano, Anna Maria; Maffei, Rossana; Bettelli, Francesca; Giusti, Davide; Messerotti, Andrea; Gilioli, Andrea; Pioli, Valeria; Leonardi, Giovanna; Forghieri, Fabio; Bresciani, Paola; Cuoghi, Angela; Morselli, Monica; Manfredini, Rossella; Longo, Giuseppe; Candoni, Anna; Marasca, Roberto; Potenza, Leonardo; Tagliafico, Enrico; Trenti, Tommaso; Comoli, Patrizia; Luppi, Mario; Riva, Giovanni
abstract

: Multiple Myeloma (MM) typically originates from underlying precursor conditions, known as Monoclonal Gammopathy of Undetermined Significance (MGUS) and Smoldering Multiple Myeloma (SMM). Validated risk factors, related to the main features of the clonal plasma cells, are employed in the current prognostic models to assess long-term probabilities of progression to MM. In addition, new prognostic immunologic parameters, measuring protective MM-specific T-cell responses, could help to identify patients with shorter time-to-progression. In this report, we described a novel Multi-antigenic Myeloma-specific (MaMs) T-cell assay, based on ELISpot technology, providing simultaneous evaluation of T-cell responses towards ten different MM-associated antigens. When performed during long-term follow-up (mean 28 months) of 33 patients with either MGUS or SMM, such deca-antigenic myeloma-specific immunoassay allowed to significantly distinguish between stable vs. progressive disease (p < 0.001), independently from the Mayo Clinic risk category. Here, we report the first clinical experience showing that a wide (multi-antigen), standardized (irrespective to patients' HLA), MM-specific T-cell assay may routinely be applied, as a promising prognostic tool, during the follow-up of MGUS/SMM patients. Larger studies are needed to improve the antigenic panel and further explore the prognostic value of MaMs test in the risk assessment of patients with monoclonal gammopathies.

2023 - Prospective evaluation of minimal residual disease in the phase II FORTE trial: a head-to-head comparison between multiparameter flow cytometry and next-generation sequencing [Articolo su rivista]
Oliva, Stefania; Genuardi, Elisa; Paris, Laura; D'Agostino, Mattia; Rogers, Jennifer; Rota-Scalabrini, Delia; Jacob, Allison P; Patriarca, Francesca; Luppi, Mario; Bertazzoni, Paola; Velluti, Cristina; Capra, Andrea; Saraci, Elona; Rossi, Marco; Allegra, Alessandro; Mina, Roberto; Gentile, Massimo; Kirsch, Ilan R; Belotti, Angelo; Cavo, Michele; Bruno, Benedetto; Musto, Pellegrino; Boccadoro, Mario; Zamagni, Elena; Gay, Francesca
abstract

Background: Limited data are available on the concordance between multiparameter flow cytometry (MFC) and next-generation sequencing (NGS) for minimal residual disease (MRD) detection in a large trial for multiple myeloma (MM) patients. Methods: MRD was explored in the FORTE trial for transplant-eligible MM patients randomised to three carfilzomib-based induction-intensification-consolidation treatments and carfilzomib-lenalidomide (KR) vs R maintenance. MRD was assessed by 8-colour 2nd-generation flow cytometry in patients with ≥very good partial response before maintenance. NGS was performed in case of suspected complete response (CR) in a correlative subanalysis. Biological/prognostic concordance between MFC and NGS, conversion to MRD negativity during maintenance, and 1-year/2-year sustained MRD negativity were explored. Findings: Between September 28, 2015 and December 22, 2021, 2020 samples were available for MFC and 728 for the simultaneous MFC/NGS correlation in the "suspected CR population". Median follow-up was 62 months. Biological agreement was 87% at the 10-5 and 83% at the 10-6 cut-offs. A remarkable prognostic concordance was observed: hazard ratios in MFC-MRD and NGS-MRD-negative vs -positive patients were 0.29 and 0.27 for progression-free survival (PFS) and 0.35 and 0.31 for overall survival, respectively (p < 0.05). During maintenance, 4-year PFS was 91% and 97% in 1-year sustained MFC-MRD-negative and NGS-MRD-negative patients (10-5), respectively, and 99% and 97% in 2-year sustained MFC-MRD-negative and NGS-MRD-negative patients, regardless of treatment received. The conversion rate from pre-maintenance MRD positivity to negativity during maintenance was significantly higher with KR vs R both by MFC (46% vs 30%, p = 0.046) and NGS (56% vs 30%, p = 0.046). Interpretation: The significant biological/clinical concordance between MFC and NGS at the same sensitivity suggests their possible use in the evaluation of one of the currently strongest predictors of outcome. Funding: Amgen, Celgene/Bristol Myers Squibb, Multiple Myeloma Research Foundation.

2023 - Prospective multicenter study on infectious complications and clinical outcome of 230 unfit acute myeloid leukemia patients receiving first-line therapy with hypomethylating agents alone or in combination with Venetoclax [Articolo su rivista]
Candoni, A.; Lazzarotto, D.; Papayannidis, C.; Piccini, M.; Nadali, G.; Dargenio, M.; Riva, M.; Fracchiolla, N.; Mellillo, L.; Dragonetti, G.; Del Principe, M. I.; Cattaneo, C.; Stulle, M.; Pasciolla, C.; De Marchi, R.; Delia, M.; Tisi, M. C.; Bonuomo, V.; Sciume, M.; Spadea, A.; Sartor, C.; Griguolo, D.; Buzzatti, E.; Basilico, C. M.; Sarlo, C.; Piccioni, A. L.; Cerqui, E.; Lessi, F.; Olivieri, A.; Fanin, R.; Luppi, M.; Pagano, L.
abstract

2023 - Role of Notch2 pathway in mature B cell malignancies [Articolo su rivista]
Mesini, N.; Fiorcari, S.; Atene, C. G.; Maffei, R.; Potenza, L.; Luppi, M.; Marasca, R.
abstract

In recent decades, the Notch pathway has been characterized as a key regulatory signaling of cell-fate decisions evolutionarily conserved in many organisms and different tissues during lifespan. At the same time, many studies suggest a link between alterations of this signaling and tumor genesis or progression. In lymphopoiesis, the Notch pathway plays a fundamental role in the correct differentiation of T and B cells, but its deregulated activity leads to leukemic onset and evolution. Notch and its ligands Delta/Jagged exhibit a pivotal role in the crosstalk between leukemic cells and their environment. This review is focused in particular on Notch2 receptor activity. Members of Notch2 pathway have been reported to be mutated in Chronic Lymphocytic Leukemia (CLL), Splenic Marginal Zone Lymphoma (SMZL) and Nodal Marginal Zone Lymphoma (NMZL). CLL is a B cell malignancy in which leukemic clones establish supportive crosstalk with non-malignant cells of the tumor microenvironment to grow, survive, and resist even the new generation of drugs. SMZL and NMZL are indolent B cell neoplasms distinguished by a distinct pattern of dissemination. In SMZL leukemic cells affect mainly the spleen, bone marrow, and peripheral blood, while NMZL has a leading nodal distribution. Since Notch2 is involved in the commitment of leukemic cells to the marginal zone as a major regulator of B cell physiological differentiation, it is predominantly affected by the molecular lesions found in both SMZL and NMZL. In light of these findings, a better understanding of the Notch receptor family pathogenic role, in particular Notch2, is desirable because it is still incomplete, not only in the physiological development of B lymphocytes but also in leukemia progression and resistance. Several therapeutic strategies capable of interfering with Notch signaling, such as monoclonal antibodies, enzyme or complex inhibitors, are being analyzed. To avoid the unwanted multiple “on target” toxicity encountered during the systemic inhibition of Notch signaling, the study of an appropriate pharmaceutical formulation is a pressing need. This is why, to date, there are still no Notch-targeted therapies approved. An accurate analysis of the Notch pathway could be useful to drive the discovery of new therapeutic targets and the development of more effective therapies.

2023 - Stigma of Palliative Care among Patients with Advanced Cancer and Their Caregivers on Early Palliative Care [Articolo su rivista]
Bandieri, Elena; Borelli, Eleonora; Gilioli, Fabio; Bigi, Sarah; Mucciarini, Claudia; Ferrari, Umberto; Eliardo, Sonia; Pinto, Lidia; Porro, Carlo Adolfo; Efficace, Fabio; Luppi, Mario; Potenza, Leonardo
abstract

: The early referral to palliative care (PC) represents a successful value-based model with proven benefits regarding the quality of life and clinical outcomes for advanced cancer patients and their caregivers. Yet, its provision remains typically confined to the last weeks of life as per the historical, late PC model. The stigma according to which PC represents end-of-life care has been identified as the root of the problem. To explore the presence and effects of the stigma in a clinical context, we surveyed 78 patients and 110 caregivers (mean age: 71.7 and 60.7, respectively) on early PC to study what their perception of PC was before their direct experience. The responses were analyzed through a qualitative descriptive approach. The participants explicitly mentioned a lack of knowledge about PC (53% of the sample), which they identified also among physicians and the population (13%); an identification of PC with the late PC model (53%); and a detrimental reaction to the proposal of an early PC referral (83%). However, the participants explicitly mentioned that a direct experience of early PC allowed for an acquired awareness of early PC meaning and benefits (52%), as well as a comprehension of its differences with late PC (34%); the regret for the delayed referral (8%); the perception of the word "palliative" as a barrier (21%); and the belief that early PC should be part of the cancer routine practice (25%). A comprehensive multi-level intervention is necessary for a widespread understanding of the essence of anticipated PC.

2023 - Vancomycin resistant enterococcus risk factors for hospital colonization in hematological patients: a matched case-control study [Articolo su rivista]
Meschiari, Marianna; Kaleci, Shaniko; Monte, Martina Del; Dessilani, Andrea; Santoro, Antonella; Scialpi, Francesco; Franceschini, Erica; Orlando, Gabriella; Cervo, Adriana; Monica, Morselli; Forghieri, Fabio; Venturelli, Claudia; Ricchizzi, Enrico; Chester, Johanna; Sarti, Mario; Guaraldi, Giovanni; Luppi, Mario; Mussini, Cristina
abstract

BackgroundVancomycin-resistant enterococcus (VRE) was the fastest growing pathogen in Europe in 2022 (+ 21%) but its clinical relevance is still unclear. We aim to identify risk factors for acquired VRE rectal colonization in hematological patients and evaluate the clinical impact of VRE colonization on subsequent infection, and 30- and 90-day overall mortality rates, compared to a matched control group.MethodsA retrospective, single center, case-control matched study (ratio 1:1) was conducted in a hematological department from January 2017 to December 2020. Case patients with nosocomial isolation of VRE from rectal swab screening (>= 48 h) were matched to controls by age, sex, ethnicity, and hematologic disease. Univariate and multivariate logistic regression compared risk factors for colonization.ResultsA total of 83 cases were matched with 83 controls. Risk factors for VRE colonization were febrile neutropenia, bone marrow transplant, central venous catheter, bedsores, reduced mobility, altered bowel habits, cachexia, previous hospitalization and antibiotic treatments before and during hospitalization. VRE bacteraemia and Clostridioides difficile infection (CDI) occurred more frequently among cases without any impact on 30 and 90-days overall mortality. Vancomycin administration and altered bowel habits were the only independent risk factors for VRE colonization at multivariate analysis (OR: 3.53 and 3.1; respectively).ConclusionsAntimicrobial stewardship strategies to reduce inappropriate Gram-positive coverage in hematological patients is urgently required, as independent risk factors for VRE nosocomial colonization identified in this study include any use of vancomycin and altered bowel habits. VRE colonization and infection did not influence 30- and 90-day mortality. There was a strong correlation between CDI and VRE, which deserves further investigation to target new therapeutic approaches.

2023 - Words hurt: common and distinct neural substrates underlying nociceptive and semantic pain [Articolo su rivista]
Borelli, Eleonora; Benuzzi, Francesca; Ballotta, Daniela; Bandieri, Elena; Luppi, Mario; Cacciari, Cristina; Porro, Carlo Adolfo; Lui, Fausta
abstract

Introduction: Recent studies have shown that processing semantic pain, such as words associated with physical pain, modulates pain perception and enhances activity in regions of the pain matrix. A direct comparison between activations due to noxious stimulation and processing of words conveying physical pain may clarify whether and to what extent the neural substrates of nociceptive pain are shared by semantic pain. Pain is triggered also by experiences of social exclusion, rejection or loss of significant others (the so-called social pain), therefore words expressing social pain may modulate pain perception similarly to what happens with words associated with physical pain. This event-related fMRI study aims to compare the brain activity related to perceiving nociceptive pain and that emerging from processing semantic pain, i.e., words related to either physical or social pain, in order to identify common and distinct neural substrates. Methods: Thirty-four healthy women underwent two fMRI sessions each. In the Semantic session, participants were presented with positive words, negative pain-unrelated words, physical pain-related words, and social pain-related words. In the Nociceptive session, participants received cutaneous mechanical stimulations that could be either painful or not. During both sessions, participants were asked to rate the unpleasantness of each stimulus. Linguistic stimuli were also rated in terms of valence, arousal, pain relatedness, and pain intensity, immediately after the Semantic session. Results: In the Nociceptive session, the 'nociceptive stimuli' vs. 'non-nociceptive stimuli' contrast revealed extensive activations in SI, SII, insula, cingulate cortex, thalamus, and dorsolateral prefrontal cortex. In the Semantic session, words associated with social pain, compared to negative pain-unrelated words, showed increased activity in most of the same areas, whereas words associated with physical pain, compared to negative pain-unrelated words, only activated the left supramarginal gyrus and partly the postcentral gyrus. Discussion: Our results confirm that semantic pain partly shares the neural substrates of nociceptive pain. Specifically, social pain-related words activate a wide network of regions, mostly overlapping with those pertaining to the affective-motivational aspects of nociception, whereas physical pain-related words overlap with a small cluster including regions related to the sensory-discriminative aspects of nociception. However, most regions of overlap are differentially activated in different conditions.

2022 - Acute promyelocytic leukaemia long-term survivors: Higher fatigue and greater overall symptom burden [Articolo su rivista]
Sommer, K.; Vignetti, M.; Cottone, F.; Breccia, M.; Annibali, O.; Luppi, M.; Intermesoli, T.; Borlenghi, E.; Carluccio, P.; Rodeghiero, F.; Fabbiano, F.; Romani, C.; Sborgia, M.; Martino, B.; Crugnola, M.; Efficace, F.
abstract

Objective We aimed to investigate the association of fatigue with severity of other key cancer symptoms, as well as symptom interference with daily activities and outlook on life, in long-term survivors of acute promyelocytic leukaemia (APL). Methods The study sample consisted of APL survivors (n=244), with a median time from diagnosis of 14.3 years (IQR=11.1-16.9 years), previously enrolled in a long-term follow-up study. Symptom severity and symptom interference were assessed using the well-validated MD Anderson Symptom Inventory (MDASI). Fatigue was evaluated with the Functional Assessment of Chronic Illness Therapy-Fatigue questionnaire. Results Higher fatigue burden was associated with increased affective symptoms, memory problems, drowsiness, sleep disturbances, shortness of breath and pain. Higher levels of fatigue were also associated with higher scores across all interference items of the MDASI. Overall, symptoms interfered most with mood, but among APL survivors with high levels of fatigue, symptoms interfered most with enjoyment of life. Multivariable regression analysis confirmed the independent association between fatigue and all symptom severity items of the MDASI. Conclusions The current findings show that long-term APL survivors who report higher fatigue also experience a greater overall symptom burden and a substantial impact on performance of daily activities. Further studies are needed to examine whether interventions aimed at reducing fatigue could also reduce overall symptom burden.

2022 - Adverse outcome associated with daratumumab-based treatments in relapsed/refractory multiple myeloma patients with amplification of chromosome arm 1q21: a single-center retrospective experience [Articolo su rivista]
Barbieri, Emiliano; Maccaferri, Monica; Leonardi, Giovanna; Giacobbi, Francesca; Corradini, Giorgia; Lagreca, Ivana; Barozzi, Patrizia; Potenza, Leonardo; Marasca, Roberto; Luppi, Mario
abstract

2022 - An 81-Year-Old Man with a 6-Year History of Chronic Lymphocytic Leukemia Presenting with Disease Flare Following Ibrutinib Discontinuation [Articolo su rivista]
Pozzi, S.; Potenza, L.; Giusti, D.; Colaci, E.; Pioli, V.; Leonardi, G.; Maccaferri, M.; Luppi, M.; Marasca, R.
abstract

Patient: Male, 81-year-old Final Diagnosis: Chronic lymphocytic leukemia Symptoms: Fever Medication: — Clinical Procedure: — Specialty: Hematology Objective: Background: Case Report: Conclusions: Unusual clinical course Chronic lymphocytic leukemia (CLL) is a mature B-cell neoplasm and the most common leukemia in adults in Western countries. Novel agents, including BTK inhibitors and the BCL2 inhibitor venetoclax, have dramati-cally changed the treatment landscape. Moreover, a disease flare, characterized by sudden worsening of clinical symptoms, radiographic findings of rapidly worsening splenomegaly or lymphadenopathy, and laboratory changes (increased absolute lymphocyte count or lactate dehydrogenase), is a phenomenon described in up to 25% of patients with CLL after ibrutinib discontinuation. We describe a patient with CLL with disease flare after ibrutinib discontinuation due to disease progression and describe the subsequent management of vene-toclax initial treatment in the course of the disease flare. We describe the case of an 81-year-old man with a 6-year history of CLL who was treated with multiple lines of therapy and developed worsening of disease-related signs and symptoms with fever, marked increase of lym-phocyte count, acute worsening of renal function, and increase in lymph nodes and spleen size following ces-sation of targeted therapy with ibrutinib at the time of disease progression. There was subsequent overlap-ping of ibrutinib during the venetoclax dose escalation period to prevent disease flare recurrence. Our report highlights the problem of disease flare after ibrutinib discontinuation in order to avoid associated patient morbidity, underscoring the importance of awareness of this phenomenon and focusing on the addition of venetoclax at time of progression in ibrutinib-treated patients, as a temporary overlap strategy, to prevent disease flare.

2022 - BTK Inhibitors Impair Platelet-Mediated Antifungal Activity [Articolo su rivista]
Nasillo, V.; Lagreca, I.; Vallerini, D.; Barozzi, P.; Riva, G.; Maccaferri, M.; Paolini, A.; Forghieri, F.; Fiorcari, S.; Maffei, R.; Martinelli, S.; Atene, C. G.; Castelli, I.; Marasca, R.; Potenza, L.; Comoli, P.; Manfredini, R.; Tagliafico, E.; Trenti, T.; Luppi, M.
abstract

In recent years, the introduction of new drugs targeting Bruton’s tyrosine kinase (BTK) has allowed dramatic improvement in the prognosis of patients with chronic lymphocytic leukemia (CLL) and other B-cell neoplasms. Although these small molecules were initially considered less immunosuppressive than chemoimmunotherapy, an increasing number of reports have described the occurrence of unexpected opportunistic fungal infections, in particular invasive aspergillosis (IA). BTK represents a crucial molecule in several signaling pathways depending on different immune receptors. Based on a variety of specific off-target effects on innate immunity, namely on neutrophils, monocytes, pulmonary macrophages, and nurse-like cells, ibrutinib has been proposed as a new host factor for the definition of probable invasive pulmonary mold disease. The role of platelets in the control of fungal growth, through granule-dependent mechanisms, was described in vitro almost two decades ago and is, so far, neglected by experts in the field of clinical management of IA. In the present study, we confirm the antifungal role of platelets, and we show, for the first time, that the exposure to BTK inhibitors impairs several immune functions of platelets in response to Aspergillus fumigatus, i.e., the ability to adhere to conidia, activation (as indicated by reduced expression of P-selectin), and direct killing activity. In conclusion, our experimental data suggest that antiplatelet effects of BTK inhibitors may contribute to an increased risk for IA in CLL patients.

2022 - Brentuximab vedotin consolidation after autologous stem cell transplantation for Hodgkin lymphoma: A Fondazione Italiana Linfomi real-life experience [Articolo su rivista]
Massaro, F.; Pavone, V.; Stefani, P. M.; Botto, B.; Pulsoni, A.; Patti, C.; Cantonetti, M.; Visentin, A.; Scalzulli, P. R.; Rossi, A.; Galimberti, S.; Cimminiello, M.; Gini, G.; Musso, M.; Sorio, M.; Arcari, A.; Zilioli, V. R.; Luppi, M.; Mannina, D.; Fabbri, A.; Pietrantuono, G.; Annibali, O.; Tafuri, A.; Prete, E.; Mule, A.; Barbolini, E.; Marcheselli, L.; Luminari, S.; Merli, F.
abstract

The standard management for relapsed or refractory classical Hodgkin lymphoma (cHL) is salvage therapy followed by autologous stem cell transplantation (ASCT). This strategy allows almost 50% of patients to be cured. Post-ASCT maintenance treatment with brentuximab vedotin (BV) confers improved progression-free survival (PFS) to cHL patients at high risk of relapse. We investigated the outcome of 105 cHL patients receiving post-ASCT BV maintenance in the real-life setting of 23 Italian hematology centers. This population included naïve patients and those previously exposed to BV. Median follow-up was 20 months. Patients presented a median of two lines of treatment pre-ASCT, with 51% receiving BV. Twenty-nine percent of patients had at least two high-risk factors (refractory disease, complete response [CR] less than 12 months, extranodal disease at relapse), while 16% presented none. At PET-CT, a Deauville score (DS) of 1–3 was reported in 75% and 78% of pre- and post-ASCT evaluations, respectively. Grade 3–4 adverse events (AEs), mainly peripheral neuropathy, were observed in 16% of patients. Three-year PFS and overall survival (OS) were 62% and 86%, respectively. According to BV exposure, 3-year PFS and OS were 54% and 71%, respectively, for naïve and 77% and 96%, respectively, for previously exposed patients. Refractory disease (hazard ratio [HR] 4.46; p = 0.003) and post-ASCT DS 4–5 (HR 3.14; p = 0.005) were the only two factors significantly associated with PFS reduction in multivariable analysis. Post-ASCT BV maintenance is an effective, safe treatment option for cHL naïve patients and those previously exposed to BV.

2022 - Chimeric Antigen Receptor T-cell Therapy in Hematologic Malignancies and Patient-reported Outcomes: A Scoping Review [Articolo su rivista]
Efficace, F.; Cannella, L.; Sparano, F.; Giesinger, J. M.; Vignetti, M.; Baron, F.; Bruera, E.; Luppi, M.; Platzbecker, U.
abstract

The inclusion of patient-reported outcome (PRO) measures in chimeric antigen receptor (CAR) T-cell therapy research is critical for understanding the impact of this novel approach from a unique patient standpoint. We performed a scoping review to map the available literature on the use of PRO measures in CAR T-cell therapy studies of patients with hematologic malignancies published between January 2015 and July 2022. Fourteen studies were identified, of which 7 (50%) were investigational early-phase trials, 6 (42.9%) were observational studies, and 1 (7.1%) was a pilot study. The EQ-5D and the PROMIS-29 were the 2 most frequently used PRO measures, being included in 6 (42.9%) and 5 (35.7%) studies, respectively. Despite differences in study designs, there seems to be evidence of improvements over time since CAR T-cell infusion in important domains such as physical functioning and fatigue, at least in patients who respond to therapy. Overall, the studies identified in our review have shown the added value of PRO assessment in CAR T-cell therapy research by providing novel information that complements the knowledge on safety and efficacy. However, there are several questions which remain to be answered in future research. For example, limited evidence exists regarding patient experience during important phases of the disease trajectory as only 4 (28.6%) and 5 (35.7%) studies provided information on PROs during the first 2 weeks from CAR T-cell infusion and after the first year, respectively. Time is ripe for a more systematic implementation of high-quality PRO assessment in future clinical trials and in real-life settings of patients treated with CAR T-cell therapy.

2022 - Constitutional Mosaicism: A Critical Issue in the Definition of BRCA-Inherited Cancer Risk [Articolo su rivista]
Tenedini, Elena; Piana, Simonetta; Toss, Angela; Marino, Marco; Barbieri, Elena; Artuso, Lucia; Venturelli, Marta; Gasparini, Elisa; Mandato, Vincenzo Dario; Marchi, Isabella; Castellano, Sara; Luppi, Mario; Trenti, Tommaso; Cortesi, Laura; Tagliafico, Enrico
abstract

2022 - ELN2017 risk stratification improves outcome prediction when applied to the prospective GIMEMA AML1310 protocol [Articolo su rivista]
Buccisano, F.; Palmieri, R.; Piciocchi, A.; Arena, V.; Candoni, A.; Melillo, L.; Calafiore, V.; Cairoli, R.; de Fabritiis, P.; Storti, G.; Salutari, P.; Lanza, F.; Martinelli, G.; Luppi, M.; Capria, S.; Maurillo, L.; Del Principe, M. I.; Paterno, G.; Consalvo, M. A. I.; Ottone, T.; Lavorgna, S.; Voso, M. T.; Fazi, P.; Vignetti, M.; Arcese, W.; Venditti, A.
abstract

The 2017 version of the European LeukemiaNet (ELN) recommendations, by integrating cytogenetics and mutational status of specific genes, divides patients with acute myeloid leukemia into 3 prognostically distinct risk categories: favorable (ELN2017-FR), intermediate (ELN2017-IR), and adverse (ELN2017-AR). We performed a post hoc analysis of the GIMEMA (Gruppo Italiano Malattie EMatologiche dell’Adulto) AML1310 trial to investigate the applicability of the ELN2017 risk stratification to our study population. In this trial, after induction and consolidation, patients in complete remission were to receive an autologous stem cell transplant (auto-SCT) if categorized as favorable risk or an allogeneic stem cell transplant (allo-SCT) if adverse risk. Intermediate-risk patients were to receive auto-SCT or allo-SCT based on the postconsolidation levels of measurable residual disease as measured by using flow cytometry. Risk categorization was originally conducted according to the 2009 National Comprehensive Cancer Network recommendations. Among 500 patients, 445 (89%) were reclassified according to the ELN2017 criteria: ELN2017-FR, 186 (41.8%) of 455; ELN2017-IR, 179 (40.2%) of 445; and ELN2017-AR, 80 (18%) of 455. In 55 patients (11%), ELN2017 was not applicable. Two-year overall survival (OS) was 68.8%, 51.3%, 45.8%, and 42.8% for the ELN2017-FR, ELN2017-IR, ELN2017-not classifiable, and ELN2017-AR groups, respectively (P, .001). When comparing the 2 different transplant strategies in each ELN2017 risk category, a significant benefit of auto-SCT over allo-SCT was observed among ELN2017-FR patients (2-year OS of 83.3% vs 66.7%; P 5 .0421). The 2 transplant procedures performed almost equally in the ELN2017-IR group (2-year OS of 73.9% vs 70.8%; P 5 .5552). This post hoc analysis of the GIMEMA AML1310 trial confirms that the ELN2017 classification is able to accurately discriminate patients with different outcomes and who may benefit from different transplant strategies. This trial was registered as EudraCT number 2010-023809-36 and at www.clinicaltrials.gov as #NCT01452646.

2022 - Early Palliative Care in Acute Myeloid Leukemia [Articolo su rivista]
Potenza, L.; Borelli, E.; Bigi, S.; Giusti, D.; Longo, G.; Odejide, O.; Porro, C. A.; Zimmermann, C.; Efficace, F.; Bruera, E.; Luppi, M.; Bandieri, E.
abstract

Background: Several novel targeted therapies seem to improve the outcome of acute myeloid leukemia (AML) patients. Nonetheless, the 5-year survival rate remains below 40%, and the trajectory of the disease remains physically and emotionally challenging, with little time to make relevant decisions. For patients with advanced solid tumors, the integration of early palliative care (EPC) with standard oncologic care a few weeks after diagnosis has demonstrated several benefits. However, this model is underutilized in patients with hematologic malignancies. Methods: In this article, we analyze the palliative care (PC) needs of AML patients, examine the operational aspects of an integrated model, and review the evidence in favor of EPC integration in the AML course. Results: AML patients have a high burden of physical and psychological symptoms and high use of avoidant coping strategies. Emerging studies, including a phase III randomized controlled trial, have reported that EPC is feasible for inpatients and outpatients, improves quality of life (QoL), promotes adaptive coping, reduces psychological symptoms, and enhances the quality of end-of-life care. Conclusions: EPC should become the new standard of care for AML patients. However, this raises issues about the urgent development of adequate programs of education to increase timely access to PC.

2022 - Edmonton symptom assessment system Global Distress Score and overall survival in acute leukaemia [Articolo su rivista]
Morselli, M.; Banchelli, F.; Borelli, E.; Cordella, S.; Forghieri, F.; Bettelli, F.; Bigi, S.; Longo, G.; D'Amico, R.; Porro, C. A.; Efficace, F.; Bruera, E.; Luppi, M.; Bandieri, E.; Potenza, L.
abstract

2022 - Edmonton symptom assessment system global distress score and overall survival among patients with advanced cancer receiving early palliative care [Articolo su rivista]
Bandieri, E.; Banchelli, F.; Borelli, E.; D'Amico, R.; Efficace, F.; Bruera, E.; Luppi, M.; Potenza, L.
abstract

2022 - GITMO Registry Study on Allogeneic Transplantation in Patients Aged ≥60 Years from 2000 to 2017: Improvements and Criticisms [Articolo su rivista]
Malagola, M.; Polverelli, N.; Rubini, V.; Martino, M.; Patriarca, F.; Bruno, B.; Giaccone, L.; Grillo, G.; Bramanti, S.; Bernasconi, P.; De Gobbi, M.; Natale, A.; Terruzzi, E.; Olivieri, A.; Chiusolo, P.; Carella, A. M.; Casini, M.; Nozzoli, C.; Mazza, P.; Bassi, S.; Onida, F.; Vacca, A.; Falcioni, S.; Luppi, M.; Iori, A. P.; Pavone, V.; Skert, C.; Carluccio, P.; Borghero, C.; Proia, A.; Selleri, C.; Sacchi, N.; Mammoliti, S.; Oldani, E.; Ciceri, F.; Russo, D.; Bonifazi, F.
abstract

Today, allogeneic stem cell transplantation (allo-SCT) can be offered to patients up to age 70 to 72 years and represents one of the most effective curative treatments for many hematologic malignancies. The primary objective of the study was to collect data from the allo-SCTs performed in Italy between 2000 and 2017 in patients aged ≥60 years to evaluate the changes in safety and efficacy outcomes, as well as their distribution and characteristics over time. The Italian Group for Bone Marrow Transplantation, Hematopoietic Stem Cells and Cell Therapy (GITMO) AlloEld study (ClinicalTrials.gov identifier NCT04469985) is a retrospective analysis of allo-SCTs performed at 30 Italian transplantation centers in older patients (age ≥60 years) between 2000 and 2017 (n = 1996). For the purpose of this analysis, patients were grouped into 3 time periods: time A, 2000 to 2005 (n = 256; 12%); time B, 2006 to 2011 (n = 584; 29%); and time C, 2012 to 2017 (n = 1156; 59%). After a median follow-up of 5.6 years, the 5-year nonrelapse mortality (NRM) remained stable (time A, 32.8%; time B, 36.2%; and time C, 35.0%; P = .5), overall survival improved (time A, 28.4%; time B, 31.8%; and time C, 37.3%; P = .012), and the cumulative incidence of relapse was reduced (time A, 45.3%; time B, 38.2%; time C, 30.0%; P < .0001). The 2-year incidence of extensive chronic graft-versus-host disease was reduced significantly (time A, 17.2%; time B, 15.8%; time C, 12.2%; P = .004). Considering times A and B together (2000 to 2011), the 2-year NRM was positively correlated with the Hematopoietic Cell Transplantation Comorbidity Index (HCT-CI) score; NRM was 25.2% in patients with an HCT-CI score of 0, 33.9% in those with a score of 1 or 2, and 36.1% in those with a score of 3 (P < .001). However, after 2012, the HCT-CI score was not significantly predictive of NRM. This study shows that the transplantation procedure in elderly patients became more effective over time. Relapse incidence remains the major problem, and strategies to prevent it are currently under investigation (eg, post-transplantation maintenance). The selection of patients aged ≥60 could be improved by combining HCT-CI and frailty assessment to better predict NRM.

2022 - Genomic and clinical findings in myeloid neoplasms with PDGFRB rearrangement [Articolo su rivista]
Di Giacomo, D.; Quintini, M.; Pierini, V.; Pellanera, F.; La Starza, R.; Gorello, P.; Matteucci, C.; Crescenzi, B.; Fiumara, P. F.; Veltroni, M.; Borlenghi, E.; Albano, F.; Forghieri, F.; Maccaferri, M.; Bettelli, F.; Luppi, M.; Cuneo, A.; Rossi, G.; Mecucci, C.
abstract

Platelet-derived growth factor receptor B (PDGFRB) gene rearrangements define a unique subgroup of myeloid and lymphoid neoplasms frequently associated with eosinophilia and characterized by high sensitivity to tyrosine kinase inhibition. To date, various PDGFRB/5q32 rearrangements, involving at least 40 fusion partners, have been reported. However, information on genomic and clinical features accompanying rearrangements of PDGFRB is still scarce. Here, we characterized a series of 14 cases with a myeloid neoplasm using cytogenetic, single nucleotide polymorphism array, and next-generation sequencing. We identified nine PDGFRB translocation partners, including the KAZN gene at 1p36.21 as a novel partner in a previously undescribed t(1;5)(p36;q33) chromosome change. In all cases, the PDGFRB recombination was the sole cytogenetic abnormality underlying the phenotype. Acquired somatic variants were mainly found in clinically aggressive diseases and involved epigenetic genes (TET2, DNMT3A, ASXL1), transcription factors (RUNX1 and CEBPA), and signaling modulators (HRAS). By using both cytogenetic and nested PCR monitoring to evaluate response to imatinib, we found that, in non-AML cases, a low dosage (100–200 mg) is sufficient to induce and maintain longstanding hematological, cytogenetic, and molecular remissions.

2022 - Gratitude among advanced cancer patients and their caregivers: The role of early palliative care [Articolo su rivista]
Borelli, E.; Bigi, S.; Potenza, L.; Gilioli, F.; Artioli, F.; Porzio, G.; Porro, C. A.; Efficace, F.; Bruera, E.; Luppi, M.; Bandieri, E.
abstract

Objective: A cancer diagnosis represents a unique trauma, given its life-threatening, multidimensional, and uncertain nature. Gratitude is a construct representing the emotional state that arises when individuals recognize that a benefit has been received as a result of someone else’s action or a spiritual entity’s intervention. Based on the positive psychological wellbeing, gratitude has been associated with improved health outcomes even in the disease setting. Thus, the models of care that foster gratitude should be adopted in the clinical context. This study aims to explore whether and how gratitude may originate in patients with advanced cancer and their caregivers undergoing early palliative care (EPC). Methods: We analyzed 251 reports from 133 patients and 118 caregivers describing their clinical experience in two EPC units. The sources of gratitude were identified and ranked based on their frequencies. Words expressing gratitude and words referring to communication and spirituality were collected by means of the Linguistic Inquiry and Word Count software and correlated. Results: In total, 123 (92.5%) of 133 patients’ and 97 (82.2%) of 118 caregivers’ reports, respectively, included explicit or implicit expressions of gratitude. Gratitude was associated specifically with successful physical symptom management, emotional support, improved attitude toward death, better information, humanity, and the familiar environment. The use of words of gratitude in patients’ reports was positively correlated with the use of words referring to communication (r =.215, p =.026) and spirituality (r =.612, p <.001). Conclusion: Our results suggest that interventions within the EPC model based on doctor–patient–caregiver communication may allow patients and caregivers to experience a feeling of gratitude, and this may represent a resource to be exploited to improve their physical and psychosocial wellbeing.

2022 - High Levels of Circulating Tumor Plasma Cells as a Key Hallmark of Aggressive Disease in Transplant-Eligible Patients With Newly Diagnosed Multiple Myeloma [Articolo su rivista]
Bertamini, L.; Oliva, S.; Rota-Scalabrini, D.; Paris, L.; More, S.; Corradini, P.; Ledda, A.; Gentile, M.; De Sabbata, G.; Pietrantuono, G.; Pascarella, A.; Tosi, P.; Curci, P.; Gilestro, M.; Capra, A.; Galieni, P.; Pisani, F.; Annibali, O.; Monaco, F.; Liberati, A. M.; Palmieri, S.; Luppi, M.; Zambello, R.; Fazio, F.; Belotti, A.; Tacchetti, P.; Musto, P.; Boccadoro, M.; Gay, F.
abstract

PURPOSE High levels of circulating tumor plasma cells (CTC-high) in patients with multiple myeloma are a marker of aggressive disease. We aimed to confirm the prognostic impact and identify a possible cutoff value of CTC-high for the prediction of progression-free survival (PFS) and overall survival (OS), in the context of concomitant risk features and minimal residual disease (MRD) achievement.METHODS. CTC were analyzed at diagnosis with two-tube single-platform flow cytometry (sensitivity 4 x 10(-5)) in patients enrolled in the multicenter randomized FORTE clinical trial (ClinicalTrials.gov identifier: NCT02203643). MRD was assessed by second-generation multipara meter flow cytometry (sensitivity 10(-5)). We tested different cutoff values in series of multivariate (MV) Cox proportional hazards regression analyses on PFS outcome and selected the value that maximized the Harrell's C-statistic. We analyzed the impact of CTC on PFS and OS in a MV analysis including baseline features and MRD negativity.RESULTS CTC analysis was performed in 401 patients; the median follow-up was 50 months (interquartile range, 45-54 months). There was a modest correlation between the percentage of CTC and bone marrow plasma cells (r = 0.38). We identified an optimal CTC cutoff of 0.07% (approximately 5 cells/mu L, C-index 0.64). In MV analysis, CTC-high versus CTC-low patients had significantly shorter PFS (hazard ratio, 2.61; 95% CI, 1.49 to 2.97, P < .001; 4-year PFS 38% v69%) and OS (hazard ratio, 2.61; 95% CI, 1.49 to 4.56; P < .001; 4-year OS 68% v92%). The CTC levels, but not the bone marrow plasma cell levels, affected the outcome. The only factor that reduced the negative impact of CTC-high was the achievement of MRD negativity (interaction P = .039).CONCLUSION In multiple myeloma, increasing levels of CTC above an optimal cutoff represent an easy-to-assess, robust, and independent high-risk factor. The achievement of MRD negativity is the most important factor that modulates their negative prognostic impact. (C) 2022 by American Society of Clinical Oncology

2022 - Indoleamine 2, 3-Dioxygenase 1 Mediates Survival Signals in Chronic Lymphocytic Leukemia via Kynurenine/Aryl Hydrocarbon Receptor-Mediated MCL1 Modulation [Articolo su rivista]
Atene, C. G.; Fiorcari, S.; Mesini, N.; Alboni, S.; Martinelli, S.; Maccaferri, M.; Leonardi, G.; Potenza, L.; Luppi, M.; Maffei, R.; Marasca, R.
abstract

The indoleamine 2,3-dioxygenase 1 (IDO1) metabolic circuitry, comprising the first tryptophan (Trp) catabolite L-kynurenine (Kyn) and the aryl hydrocarbon receptor (AHR), has emerged as a mechanism of cancer immune evasion. Here, we investigated the functional role of the IDO1/Kyn/AHR axis in chronic lymphocytic leukemia (CLL). Our data show that CLL cells expressed an active form of the IDO1 enzyme and microenvironmental stimuli can positively modulate its expression. Interferon (IFN)-γ induces IDO1 expression through the Jak/STAT1 pathway and mediates Kyn production concomitantly with Trp consumption in CLL-conditioned media, while INCB018424 (ruxolitinib), a JAK1/2 inhibitor, impaired both effects. To characterize the involvement of IDO1 in leukemic cell maintenance, we overexpressed IDO1 by vector transfection measuring enhanced resistance to spontaneous apoptosis. IDO1 pro-survival influence was confirmed by treating CLL cells with Kyn, which mediated the increase of induced myeloid leukemia cell differentiation protein (MCL1). Conversely, AHR silencing or its blockade via CH-223191 improved the apoptosis of leukemic clones and mitigated MCL1 expression. Moreover, Kyn-treated CLL cells are less affected by the pro-apoptotic effect of ABT-199 (venetoclax), while CH-223191 showed synergistic/additive cytotoxicity with this drug. Lastly, targeting directly MCL1 in CLL cells with AMG-176, we abrogate the pro-survival effect of Kyn. In conclusion, our data identify IDO1/Kyn/AHR signaling as a new therapeutic target for CLL, describing for the first time its role in CLL pathobiology.

2022 - Linking COVID-19, monocyte activation and sepsis: MDW, a novel biomarker from cytometry [Articolo su rivista]
Riva, G.; Nasillo, V.; Luppi, M.; Tagliafico, E.; Trenti, T.
abstract

2022 - Monkeypox outbreak: after COVID-19, another challenge for the hemostatic system? [Articolo su rivista]
Marietta, M.; Coluccio, V.; Luppi, M.
abstract

2022 - Myeloablative conditioning with thiotepa-busulfan-fludarabine does not improve the outcome of patients transplanted with active leukemia: final results of the GITMO prospective trial GANDALF-01 [Articolo su rivista]
Bonifazi, F.; Pavoni, C.; Peccatori, J.; Giglio, F.; Arpinati, M.; Busca, A.; Bernasconi, P.; Grassi, A.; Iori, A. P.; Patriarca, F.; Brunello, L.; Di Grazia, C.; Carella, A. M.; Cilloni, D.; Picardi, A.; Proia, A.; Santarone, S.; Sorasio, R.; Carluccio, P.; Chiusolo, P.; Cupri, A.; Luppi, M.; Nozzoli, C.; Baronciani, D.; Casini, M.; Grillo, G.; Musso, M.; Onida, F.; Palazzo, G.; Parma, M.; Tringali, S.; Vacca, A.; Vallisa, D.; Sacchi, N.; Oldani, E.; Masciulli, A.; Gheorghiu, A.; Girmenia, C.; Martino, M.; Bruno, B.; Rambaldi, A.; Ciceri, F.
abstract

The outcome of refractory/relapsed (R/R) acute leukemias is still dismal and their treatment represents an unmet clinical need. However, allogeneic transplantation (allo-HSCT) remains the only potentially curative approach in this setting. A prospective study (GANDALF-01, NCT01814488; EUDRACT:2012-004008-37) on transplantation with alternative donors had been run by GITMO using a homogeneous myeloablative conditioning regimen with busulfan, thiotepa and fludarabine while GVHD prophylaxis was stratified by donor type. The study enrolled 101 patients; 90 found an alternative donor and 87 ultimately underwent allo-HSCT. Two-year overall survival of the entire and of the transplant population (primary endpoint) were 19% and 22%, without significant differences according to disease, donor type and disease history (relapsed vs refractory patients). Two-year progression-free survival was 19% and 17% respectively. The cumulative incidences of relapse and non-relapse mortality were 49% and 33% at two years. Acute grade II-IV and chronic GVHD occurred in 23 and 10 patients. Dose intensification with a myeloablative two-alkylating regimen as sole strategy for transplanting R/R acute leukemia does seem neither to improve the outcome nor to control disease relapse. A pre-planned relapse prevention should be included in the transplant strategy in this patient population.

2022 - Need for integrating early palliative care with standard hematology care long before the allogeneic hemopoietic stem cells transplantation [Articolo su rivista]
Potenza, L.; Borelli, E.; Bigi, S.; Ganfi, V.; Luppi, M.; Bandieri, E.
abstract

2022 - Neutralizing monoclonal antibodies in haematological patients paucisymptomatic for COVID-19: The GIMEMA EMATO-0321 study [Articolo su rivista]
Marasco, V.; Piciocchi, A.; Candoni, A.; Pagano, L.; Guidetti, A.; Musto, P.; Bruna, R.; Bocchia, M.; Visentin, A.; Turrini, M.; Tucci, A.; Pilerci, S.; Fianchi, L.; Salvini, M.; Galimberti, S.; Coviello, E.; Selleri, C.; Luppi, M.; Crea, E.; Fazi, P.; Passamonti, F.; Corradini, P.
abstract

COVID-19 continues to be a relevant issue among patients with haematological malignancies (HM). Vaccines are frequently not effective in subjects on active treatment. In this multicentre retrospective study of Gruppo Italiano Malattie EMatologiche dell'Adulto (GIMEMA), we collected data from 91 paucisymptomatic HM patients treated with anti-spike neutralizing monoclonal antibodies (nMoAbs) to determine time to viral clearance, referencing it to the expected value of 28 days from an historical group of untreated paucisymptomatic patients. Secondary endpoints included rate of hospitalization, intensive care unit (ICU) admission, COVID-19 related death and safety. SARS-CoV-2 molecular swab negativity was obtained in 86 patients (95%), with a median time of 18 days (IQR 13–26; p < 0.0001). We did not find significant variations according to age, diagnosis, treatment type, vaccination status or nMoAbs type. Rate of hospitalization due to COVID-19 progression was 12% (11/91), with 2 patients (2.2%) requiring ICU admission. With a median follow-up of 2.33 months, the overall mortality was 5.5% (5/91), with 3 deaths due to COVID-19. Side effects were rare and self-limiting. Our data suggest that nMoAbs can limit the detrimental effect of immunosuppressive treatments on COVID-19 clinical progression and time to viral clearance. The original trial was registered at www.clinicaltrials.gov as #NCT04932967.

2022 - Notch2 Increases the Resistance to Venetoclax-Induced Apoptosis in Chronic Lymphocytic Leukemia B Cells by Inducing Mcl-1 [Articolo su rivista]
Fiorcari, S.; Maffei, R.; Atene, C. G.; Mesini, N.; Maccaferri, M.; Leonardi, G.; Martinelli, S.; Paolini, A.; Nasillo, V.; Debbia, G.; Potenza, L.; Luppi, M.; Marasca, R.
abstract

Chronic lymphocytic leukemia (CLL) has experienced a clinical revolution—thanks to the discovery of crucial pathogenic mechanisms. CLL is still an incurable disease due to intrinsic or acquired resistance of the leukemic clone. Venetoclax is a Bcl-2 inhibitor with a marked activity in CLL, but emerging patterns of resistance are being described. We hypothesize that intrinsic features of CLL cells may contribute to drive mechanisms of resistance to venetoclax. We analyzed the expression of Interferon Regulatory Factor 4 (IRF4), Notch2, and Mcl-1 in a cohort of CLL patients. We evaluated CLL cell viability after genetic and pharmaceutical modulation of Notch2 expression in patients harboring trisomy 12. We tested venetoclax in trisomy 12 CLL cells either silenced or not for Notch2 expression or in combination with an inhibitor of Mcl-1, AMG-176. Trisomy 12 CLL cells were characterized by low expression of IRF4 associated with high levels of Notch2 and Mcl-1. Notch2 and Mcl-1 expression determined protection of CLL cells from spontaneous and drug-induced apoptosis. Considering the involvement of Mcl-1 in venetoclax resistance, our data demonstrated a contribution of high levels of Notch2 and Mcl-1 in a reduced response to venetoclax in CLL cells carrying trisomy 12. Furthermore, reduction of Mcl-1 expression by silencing Notch2 or by treatment with AMG-176 was able to restore the response of CLL cells to venetoclax. The expression of Notch2 identifies a subset of CLL patients, mainly harboring trisomy 12, characterized by high levels of Mcl-1. This biological mechanism may compromise an effective response to venetoclax.

2022 - Perceptions of Hope Among Bereaved Caregivers of Cancer Patients Who Received Early Palliative Care: A Content and Lexicographic Analysis [Articolo su rivista]
Bigi, Sarah; Ganfi, Vittorio; Borelli, Eleonora; Potenza, Leonardo; Artioli, Fabrizio; Eliardo, Sonia; Mucciarini, Claudia; Cottafavi, Luca; Cruciani, Massimiliano; Cacciari, Cristina; Odejide, Oreofe; Porro, Carlo Adolfo; Zimmermann, Camilla; Efficace, Fabio; Bruera, Eduardo; Luppi, Mario; Bandieri, Elena
abstract

2022 - Physicians’ Perceptions of Clinical Utility of a Digital Health Tool for Electronic Patient-Reported Outcome Monitoring in Real-Life Hematology Practice. Evidence From the GIMEMA-ALLIANCE Platform [Articolo su rivista]
Efficace, F.; Patriarca, A.; Luppi, M.; Potenza, L.; Caocci, G.; Tafuri, A.; Fazio, F.; Cartoni, C.; Petrucci, M. T.; Carmosino, I.; Moia, R.; Margiotta Casaluci, G.; Boggione, P.; Colaci, E.; Giusti, D.; Pioli, V.; Sparano, F.; Cottone, F.; De Fabritiis, P.; Ardu, N. R.; Niscola, P.; Capodanno, I.; Leporace, A. P.; Pelliccia, S.; Lugli, E.; La Sala, E.; Rigacci, L.; Santopietro, M.; Fozza, C.; Siragusa, S.; Breccia, M.; Fazi, P.; Vignetti, M.
abstract

Digital health tools are increasingly being used in cancer care and may include electronic patient-reported outcome (ePRO) monitoring systems. We examined physicians’ perceptions of usability and clinical utility of a digital health tool (GIMEMA-ALLIANCE platform) for ePRO monitoring in the real-life practice of patients with hematologic malignancies. This tool allows for the collection and assessment of ePROs with real-time graphical presentation of results to medical staff. Based on a predefined algorithm, automated alerts are sent to medical staff. Participating hematologists completed an online survey on their experience with the platform. Of the 201 patients invited to participate between December 2020 and June 2021 (cut-off date for current analysis), 180 (90%) agreed to enter the platform and had a median age of 57 years. Twenty-three hematologists with a median age of 42 years and an average of 17 years of experience in clinical practice were surveyed. All hematologists agreed or strongly agreed that the platform was easy to use, and 87%, agreed or strongly agreed that ePROs data were useful to enhance communication with their patients. The majority of physicians (78%) accessed the platform at least once per month to consult the symptom and health status profile of their patients. The frequency of access was independent of physician sex (p=0.393) and years of experience in clinical practice (p=0.404). In conclusion, our preliminary results support the clinical utility, from the perspective of the treating hematologist, of integrating ePROs into the routine cancer care of patients with hematologic malignancies.

2022 - Potential mechanisms of vaccine-induced thrombosis [Articolo su rivista]
Marietta, M.; Coluccio, V.; Luppi, M.
abstract

Vaccine-induced immune thrombocytopenia and thrombosis (VITT) is a rare syndrome characterized by high-titer anti-platelet factor 4 (PF4) antibodies, thrombocytopenia and arterial and venous thrombosis in unusual sites, as cerebral venous sinuses and splanchnic veins. VITT has been described to occur almost exclusively after administration of ChAdOx1 nCoV-19 and Ad26.COV2.S adenovirus vector- based COVID-19 vaccines. Clinical and laboratory features of VITT resemble those of heparin-induced thrombocytopenia (HIT). It has been hypothesized that negatively charged polyadenylated hexone proteins of the AdV vectors could act as heparin to induce the conformational changes of PF4 molecule that lead to the formation of anti-PF4/polyanion antibodies. The anti-PF4 immune response in VITT is fostered by the presence of a proinflammatory milieu, elicited by some impurities found in ChAdOx1 nCoV-19 vaccine, as well as by soluble spike protein resulting from alternative splice events. Anti-PF4 antibodies bind PF4, forming immune complexes which activate platelets, monocytes and granulocytes, resulting in the VITT's immunothrombosis. The reason why only a tiny minority of patents receiving AdV-based COVID-19 vaccines develop VITT is still unknown. It has been hypothesized that individual intrinsic factors, either acquired (i.e., pre-priming of B cells to produce anti-PF4 antibodies by previous contacts with bacteria or viruses) or inherited (i.e., differences in platelet T-cell ubiquitin ligand-2 [TULA-2] expression) can predispose a few subjects to develop VITT. A better knowledge of the mechanistic basis of VITT is essential to improve the safety and the effectiveness of future vaccines and gene therapies using adenovirus vectors.

2022 - Secondary infections worsen the outcome of COVID-19 in patients with hematological malignancies: A report from the ITA-HEMA-COV [Articolo su rivista]
Zappasodi, P.; Cattaneo, C.; Valeria Ferretti, V.; Mina, R.; Jose Maria Ferreri, A.; Merli, F.; Oberti, M.; Krampera, M.; Romano, A.; Zerbi, C.; Ferrari, J.; Cavo, M.; Salvini, M.; Bertu, L.; Stefano Fracchiolla, N.; Marchesi, F.; Massaia, M.; Marasco, V.; Cairoli, R.; Maria Scattolin, A.; Maria Vannucchi, A.; Gambacorti-Passerini, C.; Musto, P.; Gherlinzoni, F.; Cuneo, A.; Pinto, A.; Trentin, L.; Bocchia, M.; Galimberti, S.; Coviello, E.; Chiara Tisi, M.; Morotti, A.; Falini, B.; Turrini, M.; Tafuri, A.; Billio, A.; Gentile, M.; Massimo Lemoli, R.; Venditti, A.; Giovanni Della Porta, M.; Lanza, F.; Rigacci, L.; Tosi, P.; Mohamed, S.; Corso, A.; Luppi, M.; Giuliani, N.; Busca, A.; Pagano, L.; Bruno, R.; Antonio Grossi, P.; Corradini, P.; Passamonti, F.; Arcaini, L.
abstract

The impact of secondary infections (SI) on COVID-19 outcome in patients with hematological malignancies (HM) is scarcely documented. To evaluate incidence, clinical characteristics, and outcome of SI, we analyzed the microbiologically documented SI in a large multicenter cohort of adult HM patients with COVID-19. Among 1741 HM patients with COVID-19, 134 (7.7%) had 185 SI, with a 1-month cumulative incidence of 5%. Median time between COVID-19 diagnosis and SI was 16 days (IQR: 5–36). Acute myeloid leukemia (AML) and lymphoma/plasma cell neoplasms (PCN) were more frequent diagnoses in SI patients compared to patients without SI (AML: 14.9% vs. 7.1%; lymphoma/PCN 71.7% vs. 65.3%). Patients with SI were older (median age 70 vs. 66 years, p = 0.002), with more comorbidities (median Charlson Comorbidity Index 5 vs. 4, p < 0.001), higher frequency of critical COVID-19 (19.5% vs. 11.5%, p = 0.046), and more frequently not in complete remission (75% vs. 64.7% p = 0.024). Blood and bronchoalveolar lavage were the main sites of isolation for SI. Etiology of infections was bacterial in 80% (n = 148) of cases, mycotic in 9.7% (n = 18) and viral in 10.3% (n = 19); polymicrobial infections were observed in 24 patients (18%). Escherichia coli represented most of Gram-negative isolates (18.9%), while coagulase-negative Staphylococci were the most frequent among Gram-positive (14.2%). The 30-day mortality of patients with SI was higher when compared to patients without SI (69% vs. 15%, p < 0.001). The occurrence of SI worsened COVID-19 outcome in HM patients. Timely diagnosis and adequate management should be considered to improve their prognosis.

2022 - Survey on the effectiveness of telephone-based communication with relatives of hospitalized cancer patients in COVID-19 era in Italy [Articolo su rivista]
Ricco, B.; Fiorani, C.; Ferrara, L.; Potenza, L.; Saviola, A.; Malavasi, N.; Acquaviva, G.; Carboni, C.; Scarabelli, L.; Dominici, M.; Luppi, M.; Longo, G.
abstract

Objective: No-visitor policies adopted to prevent coronavirus disease-19 (COVID-19) spread in hospital wards have deeply impacted communication with patients and their relatives. Whereas in pre-COVID-19 era family-clinician meetings were held in person, during the pandemic interactions often took place over the phone, frequently causing feelings of uncertainty and distress to the close ones at home. The goal of this study was to assess and improve the effectiveness of structured telephone-based communication with hospitalized onco-hematological patients’ relatives in COVID-19 era. Methods: After no-visitor policy was adopted in the Onco-Hematological Unit of Modena, inpatients’ relatives were contacted daily for clinical updates. After discharge, a telephone satisfaction survey was administered to all contact people of patients consecutive admitted between December 2020 and January 2021 (n = 97). Mean score of response and potential statistically significative differences depending on respondents’ characteristics were assessed. Results: Most relatives were satisfied with the communication received with a mean total score of 4.69 on a 5-point Likert scale (standard deviation: 0.60). Results showed high satisfaction rate with both the informative (mean ± SD: 4.66 ± 0.64) and emotional (mean ± SD: 4.66 ± 0.58) content, with no significant difference depending on respondents’ demographic characteristics (p > 0.05). Conclusion: A structured telephone-based communication may be a reasonable substitute for face-to-face meetings; especially if regular in time, conducted by the same doctor and integrated with video calls. Our findings might assist health workers in implementing measures to minimize the psychological effects of no-visitor policies during hospitalization. Clinical updates delivery through structured phone calls and video calls could become an opportunity also in post-COVID era.

2022 - The Response to Oxidative Damage Correlates with Driver Mutations and Clinical Outcome in Patients with Myelofibrosis [Articolo su rivista]
Genovese, E.; Mirabile, M.; Rontauroli, S.; Sartini, S.; Fantini, S.; Tavernari, L.; Maccaferri, M.; Guglielmelli, P.; Bianchi, E.; Parenti, S.; Carretta, C.; Mallia, S.; Castellano, S.; Colasante, C.; Balliu, M.; Bartalucci, N.; Palmieri, R.; Ottone, T.; Mora, B.; Potenza, L.; Passamonti, F.; Voso, M. T.; Luppi, M.; Vannucchi, A. M.; Tagliafico, E.; Manfredini, R.
abstract

Myelofibrosis (MF) is the Philadelphia-negative myeloproliferative neoplasm characterized by the worst prognosis and no response to conventional therapy. Driver mutations in JAK2 and CALR impact on JAK-STAT pathway activation but also on the production of reactive oxygen species (ROS). ROS play a pivotal role in inflammation-induced oxidative damage to cellular components including DNA, therefore leading to greater genomic instability and promoting cell transformation. In order to unveil the role of driver mutations in oxidative stress, we assessed ROS levels in CD34+ hematopoietic stem/progenitor cells of MF patients. Our results demonstrated that ROS production in CD34+ cells from CALR-mutated MF patients is far greater compared with patients harboring JAK2 mutation, and this leads to increased oxidative DNA damage. Moreover, CALR-mutant cells show less superoxide dismutase (SOD) antioxidant activity than JAK2-mutated ones. Here, we show that high plasma levels of total antioxidant capacity (TAC) correlate with detrimental clinical features, such as high levels of lactate dehydrogenase (LDH) and circulating CD34+ cells. Moreover, in JAK2-mutated patients, high plasma level of TAC is also associated with a poor overall survival (OS), and multivariate analysis demonstrated that high TAC classification is an independent prognostic factor allowing the identification of patients with inferior OS in both DIPSS lowest and highest categories. Altogether, our data suggest that a different capability to respond to oxidative stress can be one of the mechanisms underlying disease progression of myelofibrosis.

2022 - The Role of T Cell Immunity in Monoclonal Gammopathy and Multiple Myeloma: From Immunopathogenesis to Novel Therapeutic Approaches [Articolo su rivista]
Lagreca, I.; Riva, G.; Nasillo, V.; Barozzi, P.; Castelli, I.; Basso, S.; Bettelli, F.; Giusti, D.; Cuoghi, A.; Bresciani, P.; Messerotti, A.; Gilioli, A.; Pioli, V.; Colasante, C.; Vallerini, D.; Paolini, A.; Maccaferri, M.; Donatelli, F.; Forghieri, F.; Morselli, M.; Colaci, E.; Leonardi, G.; Marasca, R.; Potenza, L.; Manfredini, R.; Tagliafico, E.; Trenti, T.; Comoli, P.; Luppi, M.
abstract

Multiple Myeloma (MM) is a malignant growth of clonal plasma cells, typically arising from asymptomatic precursor conditions, namely monoclonal gammopathy of undetermined significance (MGUS) and smoldering MM (SMM). Profound immunological dysfunctions and cyto-kine deregulation are known to characterize the evolution of the disease, allowing immune escape and proliferation of neoplastic plasma cells. In the past decades, several studies have shown that the immune system can recognize MGUS and MM clonal cells, suggesting that anti-myeloma T cell immunity could be harnessed for therapeutic purposes. In line with this notion, chimeric antigen receptor T cell (CAR-T) therapy is emerging as a novel treatment in MM, especially in the re-lapsed/refractory disease setting. In this review, we focus on the pivotal contribution of T cell im-pairment in the immunopathogenesis of plasma cell dyscrasias and, in particular, in the disease progression from MGUS to SMM and MM, highlighting the potentials of T cell-based immunother-apeutic approaches in these settings.

2022 - The quality of palliative sedation in end-stage disease: audit from a department of oncology and haematology [Articolo su rivista]
Alessia, S.; Matilde, S. F.; Chrystel, I.; Massimiliano, S.; Daniele, D.; Claudia, F.; Raffaella, P.; Flavia, C.; Leonardo, F.; Chiara, C.; Leonardo, P.; Dominici, Massimo; Luppi, Mario; Giuseppe, L.
abstract

Purpose: Palliative sedation (PS) plays a critical role to give suffering relief from refractory symptoms at the end of life. Our audit aimed to assess and improve quality of PS at the Department of Oncology and Hematology of University Hospital of Modena, to verify the adherence to international guidelines, the cooperation among members of care team, focusing with attention on family’s perception of this delicate situation. Methods: From December 2016 to June 2019, data of patients undergoing PS in the Department were collected by an electronic folder tool, “Sedation Tool” (ST), that recorded clinical and PS informations, D-PaP, Rudkin score, and family’s perception. Results: In total, 245 patients were enrolled. Eighty-two percent had a Karnofsky Performance Status 10–20%. The most common cancer types were lung and gastro-intestinal carcinomas (27% and 21% respectively). Refractory symptoms observed were confusion and agitation (76%), dyspnea (39%), pain (15%), delirium (10%), and psychological distress (5%). Midazolam was the drug of choice for PS. Most of patients had Rudkin score 5 after 24 h and 33% had terminal event within a period of 24 h from the beginning of PS. During PS, most of patient’s relatives reported peacefulness (65%), agitation/impatience in 6% of cases, and concern for suffering (16%). Conclusion: PS is used in case of worsening general conditions at the end-stage disease to relieve refractory symptoms with dignity. The ST can become a simple instrument to evaluate and improve PS quality, providing more attention on the impact of PS on relatives to then possibly develop new supportive procedures for patients and their families.

2022 - Validation and functional characterization of GWAS-identified variants for chronic lymphocytic leukemia: a CRuCIAL study [Articolo su rivista]
Garcia-Martin, P.; Diez, A. M.; Maldonado, J. M. S.; Serrano, A. J. C.; ter Horst, R.; Benavente, Y.; Landi, S.; Macauda, A.; Clay-Gilmour, A.; Hernandez-Mohedo, F.; Niazi, Y.; Gonzalez-Sierra, P.; Espinet, B.; Rodriguez-Sevilla, J. J.; Maffei, R.; Blanco, G.; Giaccherini, M.; Puiggros, A.; Cerhan, J.; Marasca, R.; Canadas-Garre, M.; Lopez-Nevot, M. A.; Chen-Liang, T.; Thomsen, H.; Gamez, I.; Moreno, V.; Marcos-Gragera, R.; Garcia-Alvarez, M.; Llorca, J.; Jerez, A.; Berndt, S.; Butrym, A.; Norman, A. D.; Casabonne, D.; Luppi, M.; Slager, S. L.; Hemminki, K.; Li, Y.; Alcoceba, M.; Campa, D.; Canzian, F.; de Sanjose, S.; Forsti, A.; Netea, M. G.; Jurado, M.; Sainz, J.
abstract

2021 - A single-tube multiplex method for monitoring mutations in cysteine 481 of Bruton Tyrosine Kinase (BTK) gene in chronic lymphocytic leukemia patients treated with ibrutinib [Articolo su rivista]
Maffei, R.; Fiorcari, S.; Atene, C. G.; Martinelli, S.; Scarfo, L.; Bonfiglio, S.; Maccaferri, M.; Ljungstrom, V.; Zucchini, P.; Forghieri, F.; Potenza, L.; Ghia, P.; Marasca, R.; Trenti, T.; Tagliafico, E.; Luppi, M.
abstract

2021 - Adolescent and young adult acute lymphoblastic leukemia. Final results of the phase II pediatric-like GIMEMA LAL-1308 trial [Articolo su rivista]
Testi, A. M.; Canichella, M.; Vitale, A.; Piciocchi, A.; Guarini, A.; Starza, I. D.; Cavalli, M.; De Propris, M. S.; Messina, M.; Elia, L.; Moleti, M. L.; Martino, B.; Luppi, M.; D'Aloisio, M.; Candoni, A.; Conter, V.; Fazi, P.; Vignetti, M.; Chiaretti, S.; Foa, R.
abstract

Adolescents and young adults (AYA) with acute lymphoblastic leukemia (ALL) represent a unique patient population with specific characteristics and needs. Growing evidences suggest that pediatric-inspired approaches improve the outcome in AYA. These results prompted the design of a pediatric AIEOP-BFM ALL 2000-based regimen - the GIMEMA LAL-1308 protocol - for newly diagnosed AYA (range 18-35 years) with Philadelphia negative (Ph-) ALL. The protocol included minimal residual disease (MRD) analysis at two different time-points (TP), that is, at the end of induction IA and consolidation IB, and a modulation in post-consolidation intensity according to MRD. Seventy-six patients were eligible between September 2010 and October 2014. The regimen was well tolerated, with 2.7% induction deaths and no deaths in the post-consolidation phase. The complete response (CR) rate was 92%; the 48-month overall survival (OS) and disease-free survival (DFS) were 60.3% and 60.4%. Both OS and DFS were significantly better in T-ALL than B-ALL. A molecular MRD <10−3 at TP1 was associated with a significantly better OS and DFS (77% vs 39% and 71.9% vs 34.4%, respectively);similar results were documented at TP2 (OS and DFS 74.5% vs 30.6% and 71.5% vs 25.7%, respectively). The LAL-1308 results were compared to those from similar historic AYA populations undergoing the two previous GIMEMA LAL-2000 and LAL-0904 protocols. Both OS and DFS improved significantly compared to the two previous protocols. These results indicate that this pediatric-inspired and MRD-oriented protocol is feasible and effective for Ph- AYA ALL patients, and underline the prognostic value of MRD determinations at specific TPs.

2021 - Adoptive Transfer of JC Virus-Specific T Lymphocytes for the Treatment of Progressive Multifocal Leukoencephalopathy [Articolo su rivista]
Berzero, G.; Basso, S.; Stoppini, L.; Palermo, A.; Pichiecchio, A.; Paoletti, M.; Lucev, F.; Gerevini, S.; Rossi, A.; Vegezzi, E.; Diamanti, L.; Bini, P.; Gastaldi, M.; Delbue, S.; Perotti, C.; Seminari, E.; Faraci, M.; Luppi, M.; Baldanti, F.; Zecca, M.; Marchioni, E.; Comoli, P.
abstract

Objective: Progressive multifocal leukoencephalopathy (PML) is still burdened by high mortality in a subset of patients, such as those affected by hematological malignancies. The aim of this study was to analyze the safety and carry out preliminary evaluation of the efficacy of polyomavirus JC (JCPyV)-specific T cell therapy in a cohort of hematological patients with PML. Methods: Between 2014 and 2019, 9 patients with a diagnosis of “definite PML” according to the 2013 consensus who were showing progressive clinical deterioration received JCPyV-specific T cells. Cell lines were expanded from autologous or allogenic peripheral blood mononuclear cells by stimulation with JCPyV antigen-derived peptides. Results: None of the patients experienced treatment-related adverse events. In the evaluable patients, an increase in the frequency of circulating JCPyV-specific lymphocytes was observed, with a decrease or clearance of JCPyV viral load in cerebrospinal fluid. In responsive patients, transient appearance of punctate areas of contrast enhancement within, or close to, PML lesions was observed, which was interpreted as a sign of immune control and which regressed spontaneously without the need for steroid treatment. Six of 9 patients achieved PML control, with 5 alive and in good clinical condition at their last follow-up. Interpretation: Among other novel treatments, T cell therapy is emerging as a viable treatment option in patients with PML, particularly for those not amenable to restoration of specific immunity. Neurologists should be encouraged to refer PML patients to specialized centers to allow access to this treatment strategy. ANN NEUROL 2021;89:769–779.

2021 - Allelic HLA Matching and Pair Origin Are Favorable Prognostic Factors for Unrelated Hematopoietic Stem Cell Transplantation in Neoplastic Hematologic Diseases: An Italian Analysis by the Gruppo Italiano Trapianto di Cellule Staminali e Terapie Cellulari, Italian Bone Marrow Donor Registry, and Associazione Italiana di Immunogenetica e Biologia dei Trapianti [Articolo su rivista]
Picardi, A.; Sacchi, N.; Miotti, V.; Lorentino, F.; Oldani, E.; Rambaldi, A.; Sessa, M.; Bruno, B.; Cerno, M.; Vago, L.; Bernasconi, P.; Arcese, W.; Benedetti, F.; Pioltelli, P.; Russo, D.; Farina, L.; Fagioli, F.; Guidi, S.; Saporiti, G.; Zallio, F.; Chiusolo, P.; Borghero, C.; Papalinetti, G.; La Rocca, U.; Milone, G.; Lamparelli, T.; Carella, A. M.; Luppi, M.; Olivieri, A.; Martino, M.; Carluccio, P.; Celeghini, I.; Andreani, M.; Gallina, A. M.; Patriarca, F.; Pollichieni, S.; Mammoliti, S.; Micciche, S.; Mangione, I.; Ciceri, F.; Bonifazi, F.
abstract

HLA molecules are important for immunoreactivity in allogeneic hematopoietic stem cell transplantation (HSCT). The Gruppo Italiano Trapianto di Cellule Staminali e Terapie Cellulari, Italian Bone Marrow Donor Registry, and Associazione Italiana di Immunogenetica e Biologia dei Trapianti promoted a retrospective observational study to evaluate HLA matching and the impact of allelic HLA mismatching and non-HLA factors on unrelated Italian HSCT outcomes. From 2012 to 2015, 1788 patients were enrolled in the study. The average donor age was 29 years and the average recipient age was 49 years. As a conditioning regimen, 71% of the patients received myeloablative conditioning. For GVHD prophylaxis, 76% received either antithymocyte or anti-T lymphocyte globulin, cyclosporine A, and methotrexate. Peripheral blood was the stem cell source in 80%. The median duration of follow-up was 53 months. Regarding HLA matching, 50% of donor-recipient pairs were 10/10 matched, 38% had 1 mismatch, and 12% had 2 or more mismatches. A total of 302 pairs shared Italian origin. Four-year overall survival (OS), progression-free survival, GVHD-free relapse-free survival, and relapse rates were 49%, 40%, 22%, and 34%, respectively. The 4-year NRM was 27%, and the 100-day cumulative incidence of grade ≥II acute GVHD (aGVHD) was 26%. In multivariate analysis, 9/10 and ≤8/10 HLA allele-matched pairs were associated with worse OS (P = .04 and. 007, respectively), NRM (P = .007 and P < .0001, respectively), and grade III-IV aGVHD (P = .0001 and. 01, respectively). Moreover, the incidences of grade II-IV aGVHD (P = .001) and chronic GVHD (P = .002) were significantly lower in Italian pairs. In conclusion, 10/10 HLA matching is a favorable prognostic factor for unrelated HSCT outcome in the Italian population. Moreover, the presence of 2 HLA-mismatched loci was associated with a higher NRM (P < .0001) and grade II-IV aGVHD (P = .006) and a poorer OS (P = .001) compared with 1 HLA-mismatched locus in early or intermediate disease phases. Finally, we found that Italian donor and recipient origin is a favorable prognostic factor for GVHD occurrence.

2021 - Automated capture-based NGS workflow: one thousand patients experience in a clinical routine framework [Articolo su rivista]
Tenedini, E; Celestini, F; Iapicca, P; Marino, M; Castellano, S; Artuso, L; Biagiarelli, F; Cortesi, L; Toss, A; Barbieri, E; Roncucci, L; Pedroni, M; Manfredini, R; Luppi, M; Trenti, T; Tagliafico, E
abstract

Objectives: The Next Generation Sequencing (NGS) based mutational study of hereditary cancer genes is crucial to design tailored prevention strategies in subjects with different hereditary cancer risk. The ease of amplicon-based NGS library construction protocols contrasts with the greater uniformity of enrichment provided by capture-based protocols and so with greater chances for detecting larger genomic rearrangements and copy-number variations. Capture-based protocols, however, are characterized by a higher level of complexity of sample handling, extremely susceptible to human bias. Robotics platforms may definitely help dealing with these limits, reducing hands-on time, limiting random errors and guaranteeing process standardization.Methods: We implemented the automation of the CE-IVD SOPHiA Hereditary Cancer Solution (TM) (HCS) libraries preparation workflow by SOPHiA GENETICS on the Hamilton's STARlet platform. We present the comparison of results between this automated approach, used for more than 1,000 DNA patients' samples, and the performances of the manual protocol evaluated by SOPHiA GENETICS onto 240 samples summarized in their HCS evaluation study.Results: We demonstrate that this automated workflow achieved the same expected goals of manual setup in terms of coverages and reads uniformity, with extremely lower standard deviations among samples considering the sequencing reads mapped onto the regions of interest.Conclusions: This automated solution offers same reliable and affordable NGS data, but with the essential advantages of a flexible, automated and integrated framework, minimizing possible human errors and depicting a laboratory's walk-away scenario.

2021 - COVID-19 elicits an impaired antibody response against SARS-CoV-2 in patients with haematological malignancies [Articolo su rivista]
Passamonti, F.; Romano, A.; Salvini, M.; Merli, F.; Porta, M. G. D.; Bruna, R.; Coviello, E.; Romano, I.; Cairoli, R.; Lemoli, R.; Farina, F.; Venditti, A.; Busca, A.; Ladetto, M.; Massaia, M.; Pinto, A.; Arcaini, L.; Tafuri, A.; Marchesi, F.; Fracchiolla, N.; Bocchia, M.; Armiento, D.; Candoni, A.; Krampera, M.; Luppi, M.; Cardinali, V.; Galimberti, S.; Cattaneo, C.; La Barbera, E. O.; Mina, R.; Lanza, F.; Visani, G.; Musto, P.; Petrucci, L.; Zaja, F.; Grossi, P. A.; Bertu, L.; Pagano, L.; Corradini, P.; Derenzini, E.; Marchetti, M.; Scattolin, A. M.; Corso, A.; Tosi, P.; Gherlinzoni, F.; Passerini, C. G.; Cavo, M.; Fava, C.; Turrini, M.; Visco, C.; Zappasodi, P.; Merli, M.; Mora, B.; Vannucchi, A. M.
abstract

COVID-19 is associated with high mortality in patients with haematological malignancies (HM) and rate of seroconversion is unknown. The ITA-HEMA-COV project (NCT04352556) investigated patterns of seroconversion for SARS-CoV-2 IgG in patients with HMs. A total of 237 patients, SARS-CoV-2 PCR-positive with at least one SARS-CoV-2 IgG test performed during their care, entered the analysis. Among these, 62 (26·2%) had myeloid, 121 (51·1%) lymphoid and 54 (22·8%) plasma cell neoplasms. Overall, 69% of patients (164 of 237) had detectable IgG SARS-CoV-2 serum antibodies. Serologically negative patients (31%, 73 of 237) were evenly distributed across patients with myeloid, lymphoid and plasma cell neoplasms. In the multivariable logistic regression, chemoimmunotherapy [odds ratio (OR), 3·42; 95% confidence interval (CI), 1·04–11·21; P = 0·04] was associated with a lower rate of seroconversion. This effect did not decline after 180 days from treatment withdrawal (OR, 0·35; 95% CI: 0·11–1·13; P = 0·08). This study demonstrates a low rate of seroconversion in HM patients and indicates that treatment-mediated immune dysfunction is the main driver. As a consequence, we expect a low rate of seroconversion after vaccination and thus we suggest testing the efficacy of seroconversion in HM patients.

2021 - Changes in Cancer Patients' and Caregivers' Disease Perceptions While Receiving Early Palliative Care: A Qualitative and Quantitative Analysis [Articolo su rivista]
Borelli, Eleonora; Bigi, Sarah; Potenza, Leonardo; Eliardo, Sonia; Artioli, Fabrizio; Mucciarini, Claudia; Cottafavi, Luca; Cagossi, Katia; Razzini, Giorgia; Cruciani, Massimiliano; Pietramaggiori, Alessandra; Fantuzzi, Valeria; Lombardo, Laura; Ferrari, Umberto; Ganfi, Vittorio; Lui, Fausta; Odejide, Oreofe; Cacciari, Cristina; Porro, Carlo Adolfo; Zimmermann, Camilla; Efficace, Fabio; Bruera, Eduardo; Luppi, Mario; Bandieri, Elena
abstract

2021 - Characterization of new ATM deletion associated with hereditary breast cancer [Articolo su rivista]
Parenti, S.; Rabacchi, C.; Marino, M.; Tenedini, E.; Artuso, L.; Castellano, S.; Carretta, C.; Mallia, S.; Cortesi, L.; Toss, A.; Barbieri, E.; Manfredini, R.; Luppi, M.; Trenti, T.; Tagliafico, E.
abstract

Next-generation sequencing (NGS)-based cancer risk screening with multigene panels has become the most successful method for programming cancer prevention strategies. ATM germ-line heterozygosity has been described to increase tumor susceptibility. In particular, families carrying heterozygous germ-line variants of ATM gene have a 5-to 9-fold risk of developing breast cancer. Recent studies identified ATM as the second most mutated gene after CHEK2 in BRCA-negative patients. Nowadays, more than 170 missense variants and several truncating mutations have been identified in ATM gene. Here, we present the molecular characterization of a new ATM deletion, identified thanks to the CNV algorithm implemented in the NGS analysis pipeline. An automated workflow implementing the SOPHiA Genetics’ Hereditary Cancer Solution (HCS) protocol was used to generate NGS libraries that were sequenced on Illumina MiSeq Platform. NGS data analysis allowed us to identify a new inactivating deletion of exons 19–27 of ATM gene. The deletion was characterized both at the DNA and RNA level.

2021 - Chromosome 16 changes do not always come for good [Articolo su rivista]
Paolini, A.; Nasillo, V.; Lusenti, B.; Roncati, L.; Tagliafico, E.; Luppi, M.
abstract

2021 - Cytomegalovirus reactivation after hematopoietic stem cell transplant with CMV-IG prophylaxis: A monocentric retrospective analysis [Articolo su rivista]
Gilioli, A.; Messerotti, A.; Bresciani, P.; Cuoghi, A.; Pioli, V.; Colasante, C.; Bettelli, F.; Giusti, D.; Forghieri, F.; Potenza, L.; Donatelli, F.; Giubbolini, R.; Galassi, L.; Marasca, R.; Banchelli, F.; D'Amico, R.; Pecorari, M.; Gennari, W.; Trenti, T.; Comoli, P.; Luppi, M.; Narni, F.
abstract

Human cytomegalovirus (CMV) represents the most common viral infection after hematopoietic stem cell transplant (HSCT), mainly occurring as reactivation from latency in seropositive patients, with a different prevalence based on the extent and timing of seroconversion in a specific population. Here, we retrospectively analyzed a cohort of patients who underwent HSCT at our Institution between 2013 and 2018, all of whom were prophylactically treated with CMV-IG (Megalotect Biotest®), to define the incidence and clinical outcomes of CMV reactivation and clinically significant infection. CMV infection occurred in 69% of our patient series, mainly resulting from reactivation, and CMV clinically significant infection (CS-CMVi) occurred in 48% of prophylactically treated patients. CMV infection and CS-CMVi impacted neither on relapse incidence nor on overall survival nor on relapse-free survival. Moreover, a very low incidence of CMV end-organ disease was documented. CMV-IG used alone as prophylactic therapy after HSCT does not effectively prevent CMV reactivation.

2021 - Different semantic and affective meaning of the words associated to physical and social pain in cancer patients on early palliative/supportive care and in healthy, pain-free individuals [Articolo su rivista]
Borelli, Eleonora; Bigi, Sarah; Potenza, Leonardo; Artioli, Fabrizio; Eliardo, Sonia; Mucciarini, Claudia; Cagossi, Katia; Razzini, Giorgia; Pasqualini, Antonella; Lui, Fausta; Ferlazzo, Fabio; Cruciani, Massimiliano; Bruera, Eduardo; Efficace, Fabio; Luppi, Mario; Cacciari, Cristina; Porro, Carlo Adolfo; Bandieri, Elena
abstract

2021 - Early palliative/supportive care in acute myeloid leukaemia allows low aggression end-of-life interventions: Observational outpatient study [Articolo su rivista]
Potenza, L.; Scaravaglio, M.; Fortuna, D.; Giusti, D.; Colaci, E.; Pioli, V.; Morselli, M.; Forghieri, F.; Bettelli, F.; Messerotti, A.; Catellani, H.; Gilioli, A.; Marasca, R.; Borelli, E.; Bigi, S.; Longo, G.; Banchelli, F.; D'Amico, R.; L Back, A.; Efficace, F.; Bruera, E.; Luppi, M.; Bandieri, E.
abstract

Objectives: Early palliative supportive care has been associated with many advantages in patients with advanced cancer. However, this model is underutilised in patients with haematological malignancies. We investigated the presence and described the frequency of quality indicators for palliative care and end-of-life care in a cohort of patients with acute myeloid leukaemia receiving early palliative supportive care. Methods: This is an observational, retrospective study based on 215 patients consecutively enrolled at a haematology early palliative supportive care clinic in Modena, Italy. Comprehensive hospital chart reviews were performed to abstract the presence of well-established quality indicators for palliative care and for aggressiveness of care near the end of life. Results: 131 patients received a full early palliative supportive care intervention. All patients had at least one and 67 (51%) patients had four or more quality indicators for palliative care. Only 2.7% of them received chemotherapy in the last 14 days of life. None underwent intubation or cardiopulmonary resuscitation and was admitted to intensive care unit during the last month of life. Only 4% had either multiple hospitalisations or two or more emergency department access. Approximately half of them died at home or in a hospice. More than 40% did not receive transfusions within 7 days of death. The remaining 84 patients, considered late referrals to palliative care, demonstrated sensibly lower frequencies of the same indicators. Conclusions: Patients with acute myeloid leukaemia receiving early palliative supportive care demonstrated high frequency of quality indicators for palliative care and low rates of treatment aggressiveness at the end of life.

2021 - Education of early palliative care specialists among hematologists and oncologists to address patients’ rather than physicians’ rights [Articolo su rivista]
Potenza, L.; Luppi, M.; Borelli, E.; Bigi, S.; Bandieri, E.
abstract

2021 - Fatigue in newly diagnosed acute myeloid leukaemia: General population comparison and predictive factors [Articolo su rivista]
Oswald, L. B.; Venditti, A.; Cella, D.; Cottone, F.; Candoni, A.; Melillo, L.; Cairoli, R.; Storti, G.; Salutari, P.; Luppi, M.; Albano, F.; Martelli, M. P.; Cuneo, A.; Tafuri, A.; Trisolini, S. M.; Tieghi, A.; Fazi, P.; Vignetti, M.; Efficace, F.
abstract

Objectives: This study compared the burden of fatigue between treatment-naïve patients with newly diagnosed acute myeloid leukaemia (AML) and the general population and investigated patient factors associated with fatigue severity. Methods: Pretreatment patient-reported fatigue was assessed with the Functional Assessment of Chronic Illness Therapy-Fatigue questionnaire in a sample of 463 newly diagnosed patients with AML who were enrolled in a clinical trial. Multivariable linear regression models were used to estimate the adjusted mean differences in fatigue between patients with AML and adults from the general population (n=847) by AML disease risk categories. A clinically meaningful difference in fatigue was defined as ≥3 points. Univariable and multivariable linear regression models were used to identify sociodemographic, clinical and molecular correlates of worse fatigue in patients with AML. Results: Patients with AML reported adjusted mean fatigue scores that were 7.5 points worse than the general population (95% CI -8.6 to -6.4, p<0.001). Across AML disease risk categories, adjusted mean differences in fatigue compared with the general population ranged from 6.7 points worse (patients with favourable risk: 95% CI -8.6 to -4.8, p<0.001) to 8.9 points worse (patients with poor risk, 95% CI -10.5 to -7.2, p<0.001). Overall, 91% of patients with AML reported fatigue that was equal to or worse than the general population's median fatigue score. Higher pretreatment fatigue was independently associated with female sex, WHO performance status ≥1 and lower platelet levels. Conclusions: Patients with newly diagnosed AML reported worse fatigue than the general population, and mean differences exceeded twice the threshold for clinical significance. Our findings may help to identify patients with AML most likely to benefit from supportive care interventions to reduce fatigue.

2021 - Herpes Simplex re-activation in patients with SARS-CoV2 pneumonia: a prospective observational study. [Articolo su rivista]
Franceschini, E; Cozzi-Lepri, A; Santoro, A; Bacca, E; Lancellotti, G; Menozzi, M; Gennari, W; Meschiari, M; Bedini, A; Orlando, G; Puzzolante, C; Digaetano, M; Milic, J; Codeluppi, M; Pecorari, M; Carli, F; Cuomo, G; Alfano, G; Corradi, L; Tonelli, R; De Maria, N; Busani, S; Biagioni, E; Coloretti, I; Guaraldi, G; Sarti, M; Luppi, M; Clini, E; Girardis, M; Gyssens, I; Mussini, C.
abstract

Background: Herpes simplex 1 co-infections in patients with COVID-19 are considered relatively uncommon; some reports on re-activations in patients in intensive-care unit have been published. The aim of the study was to analyze herpetic re-activations and their clinical manifestations in hospitalized COVID-19 patients, performing HSV-1 PCR on plasma twice a week. Methods: We conducted a prospective, observational, single-center study involving 70 consecutive patients with severe/critical SARS-CoV-2 pneumonia tested for HSV-1 hospitalized at Azienda Ospedaliero-Universitaria of Modena. Results: Of these 70 patients, 21 (30.0%) showed detectable viremia and 13 (62%) had clinically relevant manifestations of HSV-1 infection corresponding to 15 events (4 pneumonia, 5 herpes labialis, 3 gingivostomatitis, one encephalitis and two hepatitis). HSV-1 positive patients were more frequently treated with steroids than HSV-1 negative patients (76.2% vs 49.0%, p 0.036) and more often underwent mechanical ventilation (IMV) (57.1% vs 22.4%, p 0.005). In the unadjusted logistic regression analysis, steroid treatment, IMV, and higher LDH were significantly associated with an increased risk of HSV1 re-activation (odds ratio 3.33, 4.61, and 16.9, respectively). The association with use of steroids was even stronger after controlling for previous use of both tocilizumab and IMV (OR=5.13, 95% CI:1.36-19.32, p=0.016). The effect size was larger when restricting to participants who were treated with high dose of steroids while there was no evidence to support an association with use of tocilizumab. Conclusions: Our study shows a high incidence of HSV-1 reactivation both virologically and clinically in patients with SARS-CoV-51 2 severe pneumonia, especially in those treated with steroids.

2021 - How to improve prognostication in acute myeloid leukemia with cbfb‐myh11 fusion transcript: Focus on the role of molecular measurable residual disease (mrd) monitoring [Articolo su rivista]
Talami, A.; Bettelli, F.; Pioli, V.; Giusti, D.; Gilioli, A.; Colasante, C.; Galassi, L.; Giubbolini, R.; Catellani, H.; Donatelli, F.; Maffei, R.; Martinelli, S.; Barozzi, P.; Potenza, L.; Marasca, R.; Trenti, T.; Tagliafico, E.; Comoli, P.; Luppi, M.; Forghieri, F.
abstract

Acute myeloid leukemia (AML) carrying inv(16)/t(16;16), resulting in fusion transcript CBFB‐MYH11, belongs to the favorable‐risk category. However, even if most patients obtain morphological complete remission after induction, approximately 30% of cases eventually relapse. While well‐established clinical features and concomitant cytogenetic/molecular lesions have been recognized to be relevant to predict prognosis at disease onset, the independent prognostic impact of measurable residual disease (MRD) monitoring by quantitative real‐time reverse transcriptase polymerase chain reaction (qRT‐PCR), mainly in predicting relapse, actually supersedes other prognostic factors. Although the ELN Working Party recently indicated that patients affected with CBFB‐MYH11 AML should have MRD assessment at informative clinical timepoints, at least after two cycles of intensive chemotherapy and after the end of treatment, several controversies could be raised, especially on the frequency of subsequent serial monitoring, the most significant MRD thresholds (most commonly 0.1%) and on the best source to be analyzed, namely, bone marrow or peripheral blood samples. Moreover, persisting low‐level MRD positivity at the end of treatment is relatively common and not predictive of relapse, provided that transcript levels remain stably below specific thresholds. Rising MRD levels suggestive of molecular relapse/progression should thus be confirmed in subsequent samples. Further prospective studies would be required to optimize postremission monitoring and to define effective MRD‐based therapeutic strategies.

2021 - IRF4 L116R mutation promotes proliferation of chronic lymphocytic leukemia B cells inducing MYC [Articolo su rivista]
Benatti, S.; Atene, C. G.; Fiorcari, S.; Mesini, N.; Martinelli, S.; Zucchini, P.; Bacchelli, F.; Maccaferri, M.; Debbia, G.; Potenza, L.; Rossi, D.; Vallisa, D.; Trentin, L.; Gaidano, G.; Luppi, M.; Marasca, R.; Maffei, R.
abstract

2021 - IRF4 modulates the response to BCR activation in chronic lymphocytic leukemia regulating IKAROS and SYK [Articolo su rivista]
Maffei, R.; Fiorcari, S.; Benatti, S.; Atene, C. G.; Martinelli, S.; Zucchini, P.; Potenza, L.; Luppi, M.; Marasca, R.
abstract

Interferon regulatory factor 4 (IRF4) is a transcriptional regulator of immune system development and function. Here, we investigated the role of IRF4 in controlling responsiveness to B-cell receptor (BCR) stimulation in chronic lymphocytic leukemia (CLL). We modulated IRF4 levels by transfecting CLL cells with an IRF4 vector or by silencing using small-interfering RNAs. Higher IRF4 levels attenuated BCR signaling by reducing AKT and ERK phosphorylation and calcium release. Conversely, IRF4 reduction improved the strength of the intracellular cascade activated by BCR engagement. Our results also indicated that IRF4 negatively regulates the expression of the spleen tyrosine kinase SYK, a crucial protein for propagation of BCR signaling, and the zinc finger DNA-binding protein IKAROS. We modulated IKAROS protein levels both by genetic manipulation and pharmacologically by treating CLL cells with lenalidomide and avadomide (IMIDs). IKAROS promoted BCR signaling by reducing the expression of inositol 5-phosphatase SHIP1. Lastly, IMIDs induced IRF4 expression, while down-regulating IKAROS and interfered with survival advantage mediated by BCR triggering, also in combination with ibrutinib. Overall, our findings elucidate the mechanism by which IRF4 tunes BCR signaling in CLL cells. Low IRF4 levels allow an efficient transmission of BCR signal throughout the accumulation of SYK and IKAROS.

2021 - Ibrutinib interferes with innate immunity in chronic lymphocytic leukemia patients during COVID-19 infection [Articolo su rivista]
Fiorcari, S.; Atene, C. G.; Maffei, R.; Debbia, G.; Potenza, L.; Luppi, M.; Marasca, R.
abstract

2021 - Increased Plasma Levels of lncRNAs LINC01268, GAS5 and MALAT1 Correlate with Negative Prognostic Factors in Myelofibrosis [Articolo su rivista]
Fantini, Sebastian; Rontauroli, Sebastiano; Sartini, Stefano; Mirabile, Margherita; Bianchi, Elisa; Badii, Filippo; Maccaferri, Monica; Guglielmelli, Paola; Ottone, Tiziana; Palmieri, Raffaele; Genovese, Elena; Carretta, Chiara; Parenti, Sandra; Mallia, Selene; Tavernari, Lara; Salvadori, Costanza; Gesullo, Francesca; Maccari, Chiara; Zizza, Michela; Grande, Alexis; Salmoiraghi, Silvia; Mora, Barbara; Potenza, Leonardo; Rosti, Vittorio; Passamonti, Francesco; Rambaldi, Alessandro; Voso, Maria Teresa; Mecucci, Cristina; Tagliafico, Enrico; Luppi, Mario; Vannucchi, Alessandro Maria; Manfredini, Rossella
abstract

: Long non-coding RNAs (lncRNAs) have been recently described as key mediators in the development of hematological malignancies. In the last years, circulating lncRNAs have been proposed as a new class of non-invasive biomarkers for cancer diagnosis and prognosis and to predict treatment response. The present study is aimed to investigate the potential of circulating lncRNAs as non-invasive prognostic biomarkers in myelofibrosis (MF), the most severe among Philadelphia-negative myeloproliferative neoplasms. We detected increased levels of seven circulating lncRNAs in plasma samples of MF patients (n = 143), compared to healthy controls (n = 65). Among these, high levels of LINC01268, MALAT1 or GAS5 correlate with detrimental clinical variables, such as high count of leukocytes and CD34+ cells, severe grade of bone marrow fibrosis and presence of splenomegaly. Strikingly, high plasma levels of LINC01268 (p = 0.0018), GAS5 (p = 0.0008) or MALAT1 (p = 0.0348) are also associated with a poor overall-survival while high levels of LINC01268 correlate with a shorter leukemia-free-survival. Finally, multivariate analysis demonstrated that the plasma level of LINC01268 is an independent prognostic variable, suggesting that, if confirmed in future in an independent patients' cohort, it could be used for further studies to design an updated classification model for MF patients.

2021 - Inflammatory microenvironment and specific t cells in myeloproliferative neoplasms: Immunopathogenesis and novel immunotherapies [Articolo su rivista]
Nasillo, V.; Riva, G.; Paolini, A.; Forghieri, F.; Roncati, L.; Lusenti, B.; Maccaferri, M.; Messerotti, A.; Pioli, V.; Gilioli, A.; Bettelli, F.; Giusti, D.; Barozzi, P.; Lagreca, I.; Maffei, R.; Marasca, R.; Potenza, L.; Comoli, P.; Manfredini, R.; Maiorana, A.; Tagliafico, E.; Luppi, M.; Trenti, T.
abstract

The Philadelphia-negative myeloproliferative neoplasms (MPNs) are malignancies of the hematopoietic stem cell (HSC) arising as a consequence of clonal proliferation driven by somatically acquired driver mutations in discrete genes (JAK2, CALR, MPL). In recent years, along with the advances in molecular characterization, the role of immune dysregulation has been achieving increasing relevance in the pathogenesis and evolution of MPNs. In particular, a growing number of studies have shown that MPNs are often associated with detrimental cytokine milieu, expansion of the monocyte/macrophage compartment and myeloid-derived suppressor cells, as well as altered functions of T cells, dendritic cells and NK cells. Moreover, akin to solid tumors and other hemato-logical malignancies, MPNs are able to evade T cell immune surveillance by engaging the PD-1/PD-L1 axis, whose pharmacological blockade with checkpoint inhibitors can successfully restore effective antitumor responses. A further interesting cue is provided by the recent discovery of the high immunogenic potential of JAK2V617F and CALR exon 9 mutations, that could be harnessed as in-triguing targets for innovative adoptive immunotherapies. This review focuses on the recent insights in the immunological dysfunctions contributing to the pathogenesis of MPNs and outlines the potential impact of related immunotherapeutic approaches.

2021 - Ivar, an interpretation‐oriented tool to manage the update and revision of variant annotation and classification [Articolo su rivista]
Castellano, S.; Cestari, F.; Faglioni, G.; Tenedini, E.; Marino, M.; Artuso, L.; Manfredini, R.; Luppi, M.; Trenti, T.; Tagliafico, E.
abstract

The rapid evolution of Next Generation Sequencing in clinical settings, and the resulting challenge of variant reinterpretation given the constantly updated information, require robust data management systems and organized approaches. In this paper, we present iVar: a freely available and highly customizable tool with a user‐friendly web interface. It represents a platform for the unified management of variants identified by different sequencing technologies. iVar accepts variant call format (VCF) files and text annotation files and elaborates them, optimizing data organization and avoiding redundancies. Updated annotations can be periodically re‐uploaded and associated with variants as historically tracked attributes, i.e., modifications can be recorded whenever an updated value is imported, thus keeping track of all changes. Data can be visualized through variant‐centered and sample‐centered interfaces. A customizable search function can be exploited to periodically check if pathogenicity‐related data of a variant has changed over time. Patient recontacting ensuing from variant reinterpretation is made easier by iVar through the effective identification of all patients present in the database carrying a specific variant. We tested iVar by uploading 4171 VCF files and 1463 annotation files, obtaining a database of 4166 samples and 22,569 unique variants. iVar has proven to be a useful tool with good performance in terms of collecting and managing data from a medium‐throughput laboratory.

2021 - MATRix–RICE therapy and autologous haematopoietic stem-cell transplantation in diffuse large B-cell lymphoma with secondary CNS involvement (MARIETTA): an international, single-arm, phase 2 trial [Articolo su rivista]
Ferreri, A. J. M.; Doorduijn, J. K.; Re, A.; Cabras, M. G.; Smith, J.; Ilariucci, F.; Luppi, M.; Calimeri, T.; Cattaneo, C.; Khwaja, J.; Botto, B.; Cellini, C.; Nassi, L.; Linton, K.; Mckay, P.; Olivieri, J.; Patti, C.; Re, F.; Fanni, A.; Singh, V.; Bromberg, J. E. C.; Cozens, K.; Gastaldi, E.; Bernardi, M.; Cascavilla, N.; Davies, A.; Fox, C. P.; Frezzato, M.; Osborne, W.; Liberati, A. M.; Novak, U.; Zambello, R.; Zucca, E.; Cwynarski, K.
abstract

Background: Secondary CNS lymphoma is a rare but potentially lethal event in patients with diffuse large B-cell lymphoma. We aimed to assess the activity and safety of an intensive, CNS-directed chemoimmunotherapy consolidated by autologous haematopoietic stem-cell transplantation (HSCT) in patients with secondary CNS lymphoma. Methods: This international, single-arm, phase 2 trial was done in 24 hospitals in Italy, the UK, the Netherlands, and Switzerland. Adults (aged 18–70 years) with histologically diagnosed diffuse large B-cell lymphoma and CNS involvement at the time of primary diagnosis or at relapse and Eastern Cooperative Oncology Group Performance Status of 3 or less were enrolled and received three courses of MATRix (rituximab 375 mg/m2, intravenous infusion, day 0; methotrexate 3·5 g/m2, the first 0·5 g/m2 in 15 min followed by 3 g/m2 in a 3 h intravenous infusion, day 1; cytarabine 2 g/m2 every 12 h, in 1 h intravenous infusions, days 2 and 3; thiotepa 30 mg/m2, 30 min intravenous infusion, day 4) followed by three courses of RICE (rituximab 375 mg/m2, day 1; etoposide 100 mg/m2 per day in 500–1000 mL over a 60 min intravenous infusion, days 1, 2, and 3; ifosfamide 5 g/m2 in 1000 mL in a 24 h intravenous infusion with mesna support, day 2; carboplatin area under the curve of 5 in 500 mL in a 1 h intravenous infusion, day 2) and carmustine–thiotepa and autologous HSCT (carmustine 400 mg/m2 in 500 mL glucose 5% solution in a 1–2 h infusion, day −6; thiotepa 5 mg/kg in saline solution in a 2 h infusion every 12 h, days −5 and −4). The primary endpoint was progression-free survival at 1 year. Overall and complete response rates before autologous HSCT, duration of response, overall survival, and safety were the secondary endpoints. Analyses were in the modified intention-to-treat population. This study is registered with ClinicalTrials.gov, NCT02329080. The trial ended after accrual completion; the database lock was Dec 31, 2019. Findings: Between March 30, 2015, and Aug 3, 2018, 79 patients were enrolled. 75 patients were assessable. 319 (71%) of the 450 planned courses were delivered. At 1 year from enrolment the primary endpoint was met, 42 patients were progression free (progression-free survival 58%; 95% CI 55–61). 49 patients (65%; 95% CI 54–76) had an objective response after MATRix–RICE, 29 (39%) of whom had a complete response. 37 patients who responded had autologous HSCT. At the end of the programme, 46 patients (61%; 95% CI 51–71) had an objective response, with a median duration of objective response of 26 months (IQR 16–37). At a median follow-up of 29 months (IQR 20–40), 35 patients were progression-free and 33 were alive, with a 2-year overall survival of 46% (95% CI 39–53). Grade 3–4 toxicity was most commonly haematological: neutropenia in 46 (61%) of 75 patients, thrombocytopenia in 45 (60%), and anaemia in 26 (35%). 79 serious adverse events were recorded in 42 (56%) patients; four (5%) of those 79 were lethal due to sepsis caused by Gram-negative bacteria (treatment-related mortality 5%; 95% CI 0·07–9·93). Interpretation: MATRix–RICE plus autologous HSCT was active in this population of patients with very poor prognosis, and had an acceptable toxicity profile. Funding: Stand Up To Cancer Campaign for Cancer Research UK, the Swiss Cancer Research foundation, and the Swiss Cancer League.

2021 - Management of patients with acute myeloid leukemia undergoing therapy with midostaurin: a focus on antifungal prophylaxis [Articolo su rivista]
Cairoli, R.; Ferrara, F.; Girmenia, C.; Luppi, M.; Pea, F.; Specchia, G.; Venditti, A.
abstract

Both therapy and prophylaxis for infectious complications during the treatment of acute myeloid leukemia (AML) have improved, although invasive fungal disease still remains a life-threatening occurrence. Accordingly, prophylactic strategies with effective and well-tolerated antifungals remain a cornerstone of management. Herein, the recent literature on antifungal prophylaxis used during the treatment of AML is reviewed, with a focus on the use in combination with midostaurin. The multikinase inhibitor midostaurin targets FMS-like tyrosine kinase 3 (FLT3) and is approved, in association with 7 + 3, for the treatment of adult patients with newly diagnosed FLT3-mutated AML. Midostaurin has been shown to extend both overall and event-free survival in AML patients with an FLT3 mutation and is now the standard of care in FLT3+ AML. Antifungal prophylaxis should be adopted during all phases of treatment in all AML patients, and the strong CYP3A4 inhibitor posaconazole is frequently the preferred agent. As midostaurin is metabolized primarily by CYP3A4, there is a potential for drug–drug interactions that requires further evaluation. At present, the available data suggest that there are no absolute contraindications for coadministration of midostaurin with posaconazole, albeit with cautious monitoring. Considering the survival advantage offered by midostarin, concomitant administration of strong CYP3A4 inhibitors should not be ruled out, although such use should be evaluated cautiously and used on a case-by-case basis only if there are no suitable alternatives. It should also be kept in mind that patients with invasive fungal infection undergoing therapy for AML with midostaurin may need prolonged antifungal therapy, which must be based on the administration of the appropriate antifungal agent.

2021 - Monocyte Distribution Width (MDW) as novel inflammatory marker with prognostic significance in COVID-19 patients [Articolo su rivista]
Riva, G.; Castellano, S.; Nasillo, V.; Ottomano, A. M.; Bergonzini, G.; Paolini, A.; Lusenti, B.; Milic, J.; De Biasi, S.; Gibellini, L.; Cossarizza, A.; Busani, S.; Girardis, M.; Guaraldi, G.; Mussini, C.; Manfredini, R.; Luppi, M.; Tagliafico, E.; Trenti, T.
abstract

Monocyte Distribution Width (MDW), a new cytometric parameter correlating with cytomorphologic changes occurring upon massive monocyte activation, has recently emerged as promising early biomarker of sepsis. Similar to sepsis, monocyte/macrophage subsets are considered key mediators of the life-threatening hyper-inflammatory disorder characterizing severe COVID-19. In this study, we longitudinally analyzed MDW values in a cohort of 87 COVID-19 patients consecutively admitted to our hospital, showing significant correlations between MDW and common inflammatory markers, namely CRP (p < 0.001), fibrinogen (p < 0.001) and ferritin (p < 0.01). Moreover, high MDW values resulted to be prognostically associated with fatal outcome in COVID-19 patients (AUC = 0.76, 95% CI: 0.66–0.87, sensitivity 0.75, specificity 0.70, MDW threshold 26.4; RR = 4.91, 95% CI: 1.73–13.96; OR = 7.14, 95% CI: 2.06–24.71). This pilot study shows that MDW can be useful in the monitoring of COVID-19 patients, as this innovative hematologic biomarker is: (1) easy to obtain, (2) directly related to the activation state of a fundamental inflammatory cell subset (i.e. monocytes, pivotal in both cytokine storm and sepsis immunopathogenesis), (3) well correlated with clinical severity of COVID-19-associated inflammatory disorder, and, in turn, (4) endowed with relevant prognostic significance. Additional studies are needed to define further the clinical impact of MDW testing in the management of COVID-19 patients.

2021 - Multiparametric flow cytometry for MRD monitoring in hematologic malignancies: Clinical applications and new challenges [Articolo su rivista]
Riva, G.; Nasillo, V.; Ottomano, A. M.; Bergonzini, G.; Paolini, A.; Forghieri, F.; Lusenti, B.; Barozzi, P.; Lagreca, I.; Fiorcari, S.; Martinelli, S.; Maffei, R.; Marasca, R.; Potenza, L.; Comoli, P.; Manfredini, R.; Tagliafico, E.; Trenti, T.; Luppi, M.
abstract

In hematologic cancers, Minimal Residual Disease (MRD) monitoring, using either molecular (PCR) or immunophenotypic (MFC) diagnostics, allows the identification of rare cancer cells, readily detectable either in the bone marrow or in the peripheral blood at very low levels, far below the limit of classic microscopy. In this paper, we outlined the state-of-the-art of MFC-based MRD detection in different hematologic settings, highlighting main recommendations and new challenges for using such a method in patients with acute leukemias or chronic hematologic neoplasms. The combination of new molecular technologies with advanced flow cytometry is progressively allowing clinicians to design a personalized therapeutic path, proportionate to the biological aggressiveness of the disease, in particular by using novel immunotherapies, in view of a modern decision-making process, based on precision medicine. Along with the evolution of immunophenotypic and molecular diagnostics, the assessment of Minimal Residual Disease (MRD) has progressively become a keystone in the clinical management of hematologic malignancies, enabling valuable post-therapy risk stratifications and guiding risk-adapted therapeutic approaches. However, specific prognostic values of MRD in different hematological settings, as well as its appropriate clinical uses (basically, when to measure it and how to deal with different MRD levels), still need further investigations, aiming to improve standardization and harmonization of MRD monitoring protocols and MRD-driven therapeutic strategies. Currently, MRD measurement in hematological neoplasms with bone marrow involvement is based on advanced highly sensitive methods, able to detect either specific genetic abnormalities (by PCRbased techniques and next-generation sequencing) or tumor-associated immunophenotypic profiles (by multiparametric flow cytometry, MFC). In this review, we focus on the growing clinical role for MFC-MRD diagnostics in hematological malignancies-from acute myeloid and lymphoblastic leukemias (AML, B-ALL and T-ALL), to chronic lymphocytic leukemia (CLL) and multiple myeloma (MM)-providing a comparative overview on technical aspects, clinical implications, advantages and pitfalls of MFC-MRD monitoring in different clinical settings.

2021 - Neoantigen-specific T-cell immune responses: The paradigm of NPM1-mutated acute myeloid leukemia [Articolo su rivista]
Forghieri, F.; Riva, G.; Lagreca, I.; Barozzi, P.; Bettelli, F.; Paolini, A.; Nasillo, V.; Lusenti, B.; Pioli, V.; Giusti, D.; Gilioli, A.; Colasante, C.; Galassi, L.; Catellani, H.; Donatelli, F.; Talami, A.; Maffei, R.; Martinelli, S.; Potenza, L.; Marasca, R.; Tagliafico, E.; Manfredini, R.; Trenti, T.; Comoli, P.; Luppi, M.
abstract

The C-terminal aminoacidic sequence from NPM1-mutated protein, absent in normal human tissues, may serve as a leukemia-specific antigen and can be considered an ideal target for NPM1-mutated acute myeloid leukemia (AML) immunotherapy. Different in silico instruments and in vitro/ex vivo immunological platforms have identified the most immunogenic epitopes from NPM1-mutated protein. Spontaneous development of endogenous NPM1-mutated-specific cytotoxic T cells has been observed in patients, potentially contributing to remission maintenance and prolonged survival. Genetically engineered T cells, namely CAR-T or TCR-transduced T cells, directed against NPM1-mutated peptides bound to HLA could prospectively represent a promising therapeutic approach. Although either adoptive or vaccine-based immunotherapies are unlikely to be highly effective in patients with full-blown leukemia, these strategies, potentially in combination with immune-checkpoint inhibitors, could be promising in maintaining remission or preemptively eradicat-ing persistent measurable residual disease, mainly in patients ineligible for allogeneic hematopoietic stem cell transplant (HSCT). Alternatively, neoantigen-specific donor lymphocyte infusion derived from healthy donors and targeting NPM1-mutated protein to selectively elicit graft-versus-leukemia effect may represent an attractive option in subjects experiencing post-HSCT relapse. Future studies are warranted to further investigate dynamics of NPM1-mutated-specific immunity and explore whether novel individualized immunotherapies may have potential clinical utility in NPM1-mutated AML patients.

2021 - Nurse-Like Cells and Chronic Lymphocytic Leukemia B Cells: A Mutualistic Crosstalk inside Tissue Microenvironments [Articolo su rivista]
Fiorcari, S.; Maffei, R.; Atene, C. G.; Potenza, L.; Luppi, M.; Marasca, R.
abstract

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in Western countries and is an example of hematological disease where cooperation between genetic defects and tumor microenvironmental interaction is involved in pathogenesis. CLL is a disease that is considered as "addicted to the host"; indeed, the crosstalk between leukemic cells and the tumor microenvironment is essential for leukemic clone maintenance supporting CLL cells' survival, proliferation, and protection from drug-induced apoptosis. CLL cells are not innocent bystanders but actively model and manipulate the surrounding microenvironment to their own advantage. Besides the different players involved in this crosstalk, nurse-like cells (NLC) resemble features related to leukemia-associated macrophages with an important function in preserving CLL cell survival and supporting an immunosuppressive microenvironment. This review provides a comprehensive overview of the role played by NLC in creating a nurturing and permissive milieu for CLL cells, illustrating the therapeutic possibilities in order to specifically target and re-educate them.

2021 - Polymorphisms within the TNFSF4 and mapkapk2 loci influence the risk of developing invasive aspergillosis: A two-stage case control study in the context of the aspbiomics consortium [Articolo su rivista]
Sanchez-Maldonado, J. M.; Moniz-Diez, A.; Ter Horst, R.; Campa, D.; Cabrera-Serrano, A. J.; Martinez-Bueno, M.; Garrido-Collado, M. P.; Hernandez-Mohedo, F.; Fernandez-Puerta, L.; Lopez-Nevot, M. A.; Cunha, C.; Gonzalez-Sierra, P. A.; Springer, J.; Lackner, M.; Alcazar-Fuoli, L.; Fianchi, L.; Aguado, J. M.; Pagano, L.; Lopez-Fernandez, E.; Clavero, E.; Potenza, L.; Luppi, M.; Moratalla, L.; Solano, C.; Sampedro, A.; Cuenca-Estrella, M.; Lass-Florl, C.; Canzian, F.; Loeffler, J.; Li, Y.; Einsele, H.; Netea, M. G.; Vazquez, L.; Carvalho, A.; Jurado, M.; Sainz, J.
abstract

Here, we assessed whether 36 single nucleotide polymorphisms (SNPs) within the TNFSF4 and MAPKAPK2 loci influence the risk of developing invasive aspergillosis (IA). We conducted a two-stage case control study including 911 high-risk patients diagnosed with hematological malignancies that were ascertained through the aspBIOmics consortium. The meta-analysis of the discovery and replication populations revealed that carriers of the TNFSF4rs7526628T/T genotype had a significantly increased risk of developing IA (p = 0.00022). We also found that carriers of the TNFSF4rs7526628T allele showed decreased serum levels of TNFSF14 protein (p = 0.0027), and that their macrophages had a decreased fungicidal activity (p = 0.048). In addition, we observed that each copy of the MAPKAPK2rs12137965G allele increased the risk of IA by 60% (p = 0.0017), whereas each copy of the MAPKAPK2rs17013271T allele was estimated to decrease the risk of developing the disease (p = 0.0029). Mechanistically, we found that carriers of the risk MAPKAPK2rs12137965G allele showed increased numbers of CD38+IgM-IgD-plasmablasts in blood (p = 0.00086), whereas those harboring two copies of the allele had decreased serum concentrations of thymic stromal lymphopoietin (p = 0.00097). Finally, we also found that carriers of the protective MAPKAPK2rs17013271T allele had decreased numbers of CD27-IgM-IgD-B cells (p = 0.00087) and significantly lower numbers of CD14+ and CD14+CD16-cells (p = 0.00018 and 0.00023). Altogether, these results suggest a role of the TNFSF4 and MAPKAPK2 genes in determining IA risk.

2021 - Pre-existing cytopenia heralding de novo acute myeloid leukemia: Uncommon presentation of NPM1-mutated AML in a single-center study [Articolo su rivista]
Galassi, L.; Colasante, C.; Bettelli, F.; Gilioli, A.; Pioli, V.; Giusti, D.; Morselli, M.; Paolini, A.; Nasillo, V.; Lusenti, B.; Colaci, E.; Donatelli, F.; Catellani, H.; Pozzi, S.; Barbieri, E.; del Rosso, M. N.; Barozzi, P.; Lagreca, I.; Martinelli, S.; Maffei, R.; Riva, G.; Tenedini, E.; Roncati, L.; Marasca, R.; Potenza, L.; Comoli, P.; Trenti, T.; Manfredini, R.; Tagliafico, E.; Luppi, M.; Forghieri, F.
abstract

2020 - Acute myeloid leukemia in patients living with HIV infection: Several questions, fewer answers [Articolo su rivista]
Forghieri, F.; Nasillo, V.; Bettelli, F.; Pioli, V.; Giusti, D.; Gilioli, A.; Mussini, C.; Tagliafico, E.; Trenti, T.; Cossarizza, A.; Maffei, R.; Barozzi, P.; Potenza, L.; Marasca, R.; Narni, F.; Luppi, M.
abstract

Both human immunodeficiency virus (HIV) infection and acute myeloid leukemia (AML) may be considered relatively uncommon disorders in the general population, but the precise incidence of AML in people living with HIV infection (PLWH) is uncertain. However, life expectancy of newly infected HIV-positive patients receiving anti-retroviral therapy (ART) is gradually increasing, rivaling that of age-matched HIV-negative individuals, so that the occurrence of AML is also expected to progressively increase. Even if HIV is not reported to be directly mutagenic, several indirect leukemogenic mechanisms, mainly based on bone marrow microenvironment disruption, have been proposed. Despite a well-controlled HIV infection under ART should no longer be considered per se a contraindication to intensive chemotherapeutic approaches, including allogeneic hematopoietic stem cell transplantation, in selected fit patients with AML, survival outcomes are still generally unsatisfactory. We discussed several controversial issues about pathogenesis and clinical management of AML in PLWH, but few evidence-based answers may currently be provided, due to the limited number of cases reported in the literature, mainly as case reports or small retrospective case series. Prospective multicenter clinical trials are warranted to more precisely investigate epidemiology and cytogenetic/molecular features of AML in PLWH, but also to standardize and further improve its therapeutic management.

2020 - BTK Inhibition Impairs the Innate Response Against Fungal Infection in Patients With Chronic Lymphocytic Leukemia [Articolo su rivista]
Fiorcari, S.; Maffei, R.; Vallerini, D.; Scarfo, L.; Barozzi, P.; Maccaferri, M.; Potenza, L.; Ghia, P.; Luppi, M.; Marasca, R.
abstract

Infections represent a cause of morbidity and mortality in patients affected by chronic lymphocytic leukemia (CLL). Introduction of new drugs in CLL clinical practice has showed impressive efficacy, in particular those targeting BTK. Among the consistent clinical data, an increasing number of reports describing the occurrence of unexpected opportunistic fungal infections has been reported during treatment with ibrutinib in the first 6 months of treatment. The reason underlying manifestations of invasive fungal infections in patients treated with ibrutinib is still under investigation. Our study aimed to understand the impact of BTK inhibition on immune response to fungal infection mediated by macrophages and CD14+ monocytic population obtained from CLL patients. Exposure to ibrutinib and acalabrutinib reduced signaling pathways activated by Aspergillus fumigatus determining an exacerbation of an immunosuppressive signature, a reduction of phagocytosis and a significant deficit in the secretion of inflammatory cytokines either in macrophages and monocytes isolated from CLL patients and healthy donors. These effects lead to a failure in completely counteracting conidia germination. In addition we investigated the biological effects of ibrutinib on monocyte counterpart in patients who were undergoing therapy. A significant impairment in cytokine secretion and a deficit of phagocytosis in circulating monocytes were detected after 3 months of treatment. Thus, our results uncover modifications in the innate response in CLL patients induced by ibrutinib that may impair the immunological response to fungal infection. BTK inhibition affects a productive immune response of CLL-associated macrophages (NLC) during Aspergillus fumigatus infection. Reduction of TNF-α secretion and phagocytosis are detected in monocytes isolated from CLL patients during ibrutinib therapy.

2020 - COVID-19 Pandemic and Cancer: The Importance of Early Palliative Care [Articolo su rivista]
Potenza, L.; Luppi, M.; Efficace, F.; Bruera, E.; Bandieri, E.
abstract

In terms of clinical and ethical situations, this narrative compares the COVID-19 pandemic to the cancer endemic and shares information that may be helpful to improve the management of both future pandemics and cancer care.

2020 - COVID-19, coagulopathy and venous thromboembolism: more questions than answers [Articolo su rivista]
Marietta, M.; Coluccio, V.; Luppi, M.
abstract

The acute respiratory illnesses caused by severe acquired respiratory syndrome corona Virus-2 (SARS-CoV-2) is a global health emergency, involving more than 8.6 million people worldwide with more than 450,000 deaths. Among the clinical manifestations of COVID-19, the disease that results from SARS-CoV-2 infection in humans, a prominent feature is a pro-thrombotic derangement of the hemostatic system, possibly representing a peculiar clinicopathologic manifestation of viral sepsis. The severity of the derangement of coagulation parameters in COVID-19 patients has been associated with a poor prognosis, and the use of low molecular weight heparin (LMWH) at doses registered for prevention of venous thromboembolism (VTE) has been endorsed by the World Health Organization and by Several Scientific societies. However, some relevant issues on the relationships between COVID-19, coagulopathy and VTE have yet to be fully elucidated. This review is particularly focused on four clinical questions: What is the incidence of VTE in COVID-19 patients? How do we frame the COVID-19 associated coagulopathy? Which role, if any, do antiphospolipid antibodies have? How do we tackle COVID-19 coagulopathy? In the complex scenario of an overwhelming pandemic, most everyday clinical decisions have to be taken without delay, although not yet supported by a sound scientific evidence. This review discusses the most recent findings of basic and clinical research about the COVID-associated coagulopathy, to foster a more thorough knowledge of the mechanisms underlying this compelling disease.

2020 - COVID-19: More than a cytokine storm [Articolo su rivista]
Riva, G.; Nasillo, V.; Tagliafico, E.; Trenti, T.; Comoli, P.; Luppi, M.
abstract

2020 - COVID-19: Room for treating T cell exhaustion? [Articolo su rivista]
Riva, G.; Nasillo, V.; Tagliafico, E.; Trenti, T.; Luppi, M.
abstract

2020 - Cancer treatment during the coronavirus disease 2019 pandemic: Do not postpone, do it! [Articolo su rivista]
Omarini, C.; Maur, M.; Luppi, G.; Narni, F.; Luppi, M.; Dominici, M.; Longo, G.; Piacentini, F.
abstract

At the end of January 2020, a novel betacoronavirus, known as severe acute respiratory syndrome coronavirus 2, progressively spread in Italy. Patients with cancer are considered more prone to infections because of the immunosuppressive status due to both malignancy and anticancer treatments. From the first Italian government restrictions (23rd February), Modena Cancer Center adopted practical health vigilance recommendations to minimise the risk of exposure to the virus without overlooking cancer management. From 23rd February to 31st March 2020, 1257 patients on active anticancer treatment for oncological or haematological malignancies attended our institution. All the staff activities were rescheduled following our practical coronavirus disease 2019 (COVID-19) guideline. During this period, we have tallied 9 cases of COVID-19 infection (0.71%) in patients with cancer and 3 cases (1.66%) in health workers. The mortality rate of our patients with cancer was 22%, consistent with the data reported in the literature. In conclusion, following our practical health vigilance recommendations, physicians should be confident in maintaining life-saving anticancer treatment without exceedingly increasing the risk of nosocomial COVID-19 infection. The high rate of mortality suggested that all patients on active anticancer treatment with flu-like symptoms have to be carefully screened for COVID-19 infection.

2020 - Clinical characteristics and outcome of west nile virus infection in patients with lymphoid neoplasms: An italian multicentre study [Articolo su rivista]
Visentin, A.; Nasillo, V.; Marchetti, M.; Ferrarini, I.; Paolini, R.; Sancetta, R.; Rigolin, G. M.; Cibien, F.; Riva, M.; Briani, C.; Marinello, S.; Piazza, F.; Gherlinzoni, F.; Krampera, M.; Bassan, R.; Cuneo, A.; Luppi, M.; Semenzato, G.; Marasca, R.; Trentin, L.
abstract

2020 - Clinical characteristics and risk factors associated with COVID-19 severity in patients with haematological malignancies in Italy: a retrospective, multicentre, cohort study [Articolo su rivista]
Passamonti, F.; Cattaneo, C.; Arcaini, L.; Bruna, R.; Cavo, M.; Merli, F.; Angelucci, E.; Krampera, M.; Cairoli, R.; Della Porta, M. G.; Fracchiolla, N.; Ladetto, M.; Gambacorti Passerini, C.; Salvini, M.; Marchetti, M.; Lemoli, R.; Molteni, A.; Busca, A.; Cuneo, A.; Romano, A.; Giuliani, N.; Galimberti, S.; Corso, A.; Morotti, A.; Falini, B.; Billio, A.; Gherlinzoni, F.; Visani, G.; Tisi, M. C.; Tafuri, A.; Tosi, P.; Lanza, F.; Massaia, M.; Turrini, M.; Ferrara, F.; Gurrieri, C.; Vallisa, D.; Martelli, M.; Derenzini, E.; Guarini, A.; Conconi, A.; Cuccaro, A.; Cudillo, L.; Russo, D.; Ciambelli, F.; Scattolin, A. M.; Luppi, M.; Selleri, C.; Ortu La Barbera, E.; Ferrandina, C.; Di Renzo, N.; Olivieri, A.; Bocchia, M.; Gentile, M.; Marchesi, F.; Musto, P.; Federici, A. B.; Candoni, A.; Venditti, A.; Fava, C.; Pinto, A.; Galieni, P.; Rigacci, L.; Armiento, D.; Pane, F.; Oberti, M.; Zappasodi, P.; Visco, C.; Franchi, M.; Grossi, P. A.; Bertu, L.; Corrao, G.; Pagano, L.; Corradini, P.
abstract

Background: Several small studies on patients with COVID-19 and haematological malignancies are available showing a high mortality in this population. The Italian Hematology Alliance on COVID-19 aimed to collect data from adult patients with haematological malignancies who required hospitalisation for COVID-19. Methods: This multicentre, retrospective, cohort study included adult patients (aged ≥18 years) with diagnosis of a WHO-defined haematological malignancy admitted to 66 Italian hospitals between Feb 25 and May 18, 2020, with laboratory-confirmed and symptomatic COVID-19. Data cutoff for this analysis was June 22, 2020. The primary outcome was mortality and evaluation of potential predictive parameters of mortality. We calculated standardised mortality ratios between observed death in the study cohort and expected death by applying stratum-specific mortality rates of the Italian population with COVID-19 and an Italian cohort of 31 993 patients with haematological malignancies without COVID-19 (data up to March 1, 2019). Multivariable Cox proportional hazards model was used to identify factors associated with overall survival. This study is registered with ClinicalTrials.gov, NCT04352556, and the prospective part of the study is ongoing. Findings: We enrolled 536 patients with a median follow-up of 20 days (IQR 10–34) at data cutoff, 85 (16%) of whom were managed as outpatients. 440 (98%) of 451 hospitalised patients completed their hospital course (were either discharged alive or died). 198 (37%) of 536 patients died. When compared with the general Italian population with COVID-19, the standardised mortality ratio was 2·04 (95% CI 1·77–2·34) in our whole study cohort and 3·72 (2·86–4·64) in individuals younger than 70 years. When compared with the non-COVID-19 cohort with haematological malignancies, the standardised mortality ratio was 41·3 (38·1–44·9). Older age (hazard ratio 1·03, 95% CI 1·01–1·05); progressive disease status (2·10, 1·41–3·12); diagnosis of acute myeloid leukaemia (3·49, 1·56–7·81), indolent non-Hodgin lymphoma (2·19, 1·07–4·48), aggressive non-Hodgkin lymphoma (2·56, 1·34–4·89), or plasma cell neoplasms (2·48, 1·31–4·69), and severe or critical COVID-19 (4·08, 2·73–6·09) were associated with worse overall survival. Interpretation: This study adds to the evidence that patients with haematological malignancies have worse outcomes than both the general population with COVID-19 and patients with haematological malignancies without COVID-19. The high mortality among patients with haematological malignancies hospitalised with COVID-19 highlights the need for aggressive infection prevention strategies, at least until effective vaccination or treatment strategies are available. Funding: Associazione italiana contro le leucemie, linfomi e mieloma–Varese Onlus.

2020 - Early Palliative Care: A Necessary Intervention for Patients Ineligibile to Approved Potentially Life-saving CAR T-cell Therapy [Articolo su rivista]
Potenza, L.; Luppi, M.; Efficace, F.; Bruera, E.; Bandieri, E.
abstract

2020 - Early versus delayed palliative/supportive care in advanced cancer: An observational study [Articolo su rivista]
Bandieri, E.; Banchelli, F.; Artioli, F.; Mucciarini, C.; Razzini, G.; Cruciani, M.; Potenza, L.; D'Amico, R.; Efficace, F.; Bruera, E.; Luppi, M.
abstract

Objective: The positive impact of early palliative care interventions in advanced cancer patients has so far been largely evaluated in randomised controlled trials. This study aimed at providing information on the value of early palliative/supportive care, integrated with standard oncologic care, in a real-life setting. Methods: This was a retrospective observational study of 292 advanced cancer patients consecutively admitted at Carpi Hospital in Modena, Italy, between 2014 and 2017. For the purpose of this analysis, patients were classified into two groups (early and delayed palliative/supportive care patients), and analysed for different clinical indicators. Early and delayed palliative/supportive care were classified according to the time elapsed from advanced cancer diagnosis until palliative/supportive care start. Results: A total of 200 patients (68%), with at least three visits, were included in the analyses. The frequency of chemotherapy use in the last 60 days of life was 3.4% and 24.6% in the early and delayed groups, respectively (adjusted OR=0.1; 95% CI 0.0 to 0.4; p=0.002). The estimated survival probability at 1 year was 74.5% (95% CI 65.0% to 85.4%) and 45.5% (95% CI 37.6% to 55.0%), in the early and delayed groups, respectively. Performance status, pain and all the Edmonton Symptom Assessment Scale items, assessed at baseline and at 1 to 12 weeks after the intervention, showed significant improvement over time. However, no between-group differences were found with regard to symptom outcomes. Conclusions: An earlier palliative/supportive care intervention was associated with reduced aggressiveness of therapy, in patients receiving community oncology care. Symptom burden was improved by early palliative/supportive care, independently of the timing of patient referral.

2020 - Effects of Anti-vitamin k oral anticoagulants on bone and cardiovascular health [Articolo su rivista]
Marietta, M.; Coluccio, V.; Boriani, G.; Luppi, M.
abstract

Vitamin K antagonist oral anticoagulants (VKAs) have been proven over 50 years to be highly effective and acceptably safe in many settings and are still used by millions of people worldwide. The main concern about the safety of VKAs regards the risk of bleeding, but there is accumulation evidence of their potentially negative effects beyond hemostasis. Indeed, VKAs impair the action of several Vitamin-K Dependent Proteins (VKDP), such as Bone Gla protein, Matrix Gla protein, Gas6 Protein, Periostin and Gla-Ric Protein, involved in bone and vascular metabolism, thus exerting a detrimental effect on bone and vascular health. Indeed, although the evidence regarding this issue is not compelling, it has been shown that VKAs use decreases bone mass density, increases the risk of bone fractures and accelerates the process of vascular and valvular calcification. Vascular calcification is a major concern in Chronic Kidney Disease (CKD) patients, also in absence of VKAs, because of mineral metabolism derangement, chronic inflammation and oxidative stress. Direct Oral AntiCoagulants (DOACs) do not affect VKDP involved in vascular and valvular calcification, and do not induce calcific valve degeneration in animal models, being a possible alternative to AVK for CKD patients. However, the efficacy and safety of DOACs in this population, suggested by some recent observations, requires confirmation by dedicated, randomized study. We reviewed here the effects of VKAs in bone and vascular health as compared to DOACs, in order to provide the physicians with some data useful to wisely choose the most suitable anticoagulant for every patient.

2020 - Epidemiology and clinical outcomes of latent tuberculosis infection in adults affected with acute leukemia or aplastic anemia: a retrospective single-center study [Articolo su rivista]
Bettelli, F.; Giusti, D.; Morselli, M.; Colaci, E.; Nasillo, V.; Pioli, V.; Gilioli, A.; Iotti, S.; Galassi, L.; Giubbolini, R.; Colasante, C.; Catellani, H.; Barozzi, P.; Lagreca, I.; Vallerini, D.; Maffei, R.; Franceschini, E.; Mussini, C.; Banchelli, F.; D'Amico, R.; Marasca, R.; Narni, F.; Potenza, L.; Comoli, P.; Luppi, M.; Forghieri, F.
abstract

2020 - Harnessing T Cells to Control Infections After Allogeneic Hematopoietic Stem Cell Transplantation [Articolo su rivista]
Basso, S.; Compagno, F.; Zelini, P.; Giorgiani, G.; Boghen, S.; Bergami, E.; Bagnarino, J.; Siciliano, M.; Del Fante, C.; Luppi, M.; Zecca, M.; Comoli, P.
abstract

Dramatic progress in the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from alternative sources in pediatric patients has been registered over the past decade, providing a chance to cure children and adolescents in need of a transplant. Despite these advances, transplant-related mortality due to infectious complications remains a major problem, principally reflecting the inability of the depressed host immune system to limit infection replication and dissemination. In addition, development of multiple infections, a common occurrence after high-risk allo-HSCT, has important implications for overall survival. Prophylactic and preemptive pharmacotherapy is limited by toxicity and, to some extent, by lack of efficacy in breakthrough infections. T-cell reconstitution is a key requirement for effective infection control after HSCT. Consequently, T-cell immunotherapeutic strategies to boost pathogen-specific immunity may complement or represent an alternative to drug treatments. Pioneering proof of principle studies demonstrated that the administration of donor-derived T cells directed to human herpesviruses, on the basis of viral DNA monitoring, could effectively restore specific immunity and confer protection against viral infections. Since then, the field has evolved with implementation of techniques able to hasten production, allow for selection of specific cell subsets, and target multiple pathogens. This review provides a brief overview of current cellular therapeutic strategies to prevent or treat pathogen-related complications after HSCT, research carried out to increase efficacy and safety, including T-cell production for treatment of infections in patients with virus-naïve donors, results from clinical trials, and future developments to widen adoptive T-cell therapy access in the HSCT setting.

2020 - High serum ferritin levels in newly diagnosed patients with myelodysplastic syndromes are associated with greater symptom severity [Articolo su rivista]
Caocci, G.; Vignetti, M.; Patriarca, A.; Breccia, M.; Platzbecker, U.; Palumbo, G. A.; Stauder, R.; Cottone, F.; Petranovic, D.; Voso, M. T.; Tafuri, A.; Invernizzi, R.; Caers, J.; Luppi, M.; La Nasa, G.; Niscola, P.; Efficace, F.
abstract

We examined the association between serum ferritin (SF) levels and patient-reported functional aspects and symptoms, as measured by the EORTC QLQ-C30, in newly diagnosed patients with myelodysplastic syndromes (MDS). Analysis was conducted on 497 MDS patients who were classified in two groups based on the SF value of 1000 ng/mL. Clinically relevant differences of patient-reported functional and symptom scales were evaluated and classified as small, medium and large, based on established thresholds. Multivariable linear regression analysis was performed to account for potential confounding factors. Patients with SF of ≥ 1000 ng/mL reported statistically significant and clinically relevant worse outcomes across various health domains. Dyspnea was the symptom indicating the largest difference and mean scores of patients with higher and lower SF levels were 40 and 24.3, respectively (p = 0.005), indicating a large clinically relevant difference (Δ = 15.7). Further research is needed to better understand the relationship between SF levels and specific health-related quality of life domains.

2020 - Host immune genetic variations influence the risk of developing acute myeloid leukaemia: results from the NuCLEAR consortium [Articolo su rivista]
Sanchez-Maldonado, J. M.; Campa, D.; Springer, J.; Badiola, J.; Niazi, Y.; Moniz-Diez, A.; Hernandez-Mohedo, F.; Gonzalez-Sierra, P.; Ter Horst, R.; Macauda, A.; Brezina, S.; Cunha, C.; Lackner, M.; Lopez-Nevot, M. A.; Fianchi, L.; Pagano, L.; Lopez-Fernandez, E.; Potenza, L.; Luppi, M.; Moratalla, L.; Rodriguez-Sevilla, J. J.; Fonseca, J. E.; Tormo, M.; Solano, C.; Clavero, E.; Romero, A.; Li, Y.; Lass-Florl, C.; Einsele, H.; Vazquez, L.; Loeffler, J.; Hemminki, K.; Carvalho, A.; Netea, M. G.; Gsur, A.; Dumontet, C.; Canzian, F.; Forsti, A.; Jurado, M.; Sainz, J.
abstract

The purpose of this study was to conduct a two-stage case control association study including 654 acute myeloid leukaemia (AML) patients and 3477 controls ascertained through the NuCLEAR consortium to evaluate the effect of 27 immune-related single nucleotide polymorphisms (SNPs) on AML risk. In a pooled analysis of cohort studies, we found that carriers of the IL13rs1295686A/A genotype had an increased risk of AML (PCorr = 0.0144) whereas carriers of the VEGFArs25648T allele had a decreased risk of developing the disease (PCorr = 0.00086). In addition, we found an association of the IL8rs2227307 SNP with a decreased risk of developing AML that remained marginally significant after multiple testing (PCorr = 0.072). Functional experiments suggested that the effect of the IL13rs1295686 SNP on AML risk might be explained by its role in regulating IL1Ra secretion that modulates AML blast proliferation. Likewise, the protective effect of the IL8rs2227307 SNP might be mediated by TLR2-mediated immune responses that affect AML blast viability, proliferation and chemorresistance. Despite the potential interest of these results, additional functional studies are still warranted to unravel the mechanisms by which these variants modulate the risk of AML. These findings suggested that IL13, VEGFA and IL8 SNPs play a role in modulating AML risk.

2020 - Ibrutinib Is a Newly Recognized Host Factor for the Definition of Probable Invasive Pulmonary Mold Disease, Based on Off-target Effects, Unrelated to Its B-cell Immunosuppressant Activity [Articolo su rivista]
Luppi, Mario; Forghieri, Fabio; Potenza, Leonardo
abstract

2020 - Immunomodulatory effect of ibrutinib: Reducing the barrier against fungal infections [Articolo su rivista]
Maffei, R.; Maccaferri, M.; Arletti, L.; Fiorcari, S.; Benatti, S.; Potenza, L.; Luppi, M.; Marasca, Roberto
abstract

The Bruton tyrosine kinase (BTK) inhibitor ibrutinib is increasingly used in the treatment of chronic lymphocytic leukemia (CLL). Moreover, very promising results have been reported in other B-cell malignancies, including primary central nervous system lymphoma (PCNSL). Although well-tolerated in the majority of patients, ibrutinib demonstrates in some cases troublesome toxicities, including invasive fungal infections (IFIs). In the present review, we summarize clinical manifestations of IFIs in patients treated with ibrutinib, generally characterized by an early onset, mild clinical manifestations, asymptomatic/low symptomatic pulmonary localization and high incidence of central nervous system (CNS) involvement. IFI risk appears particularly increased in patients receiving ibrutinib associated with other immune modulator agents, especially with steroids or immune-chemotherapy. Moreover, the immunomodulatory effect of ibrutinib is described, pointing the attention on the involvement of specific molecules targeted by ibrutinib in innate and adaptive response to fungal infection. Overall, the findings indicate the ibrutinib may rapidly impair innate immune cell functions, while concomitantly restoring an effective protective potential of adaptive immune compartment. A correct awareness, especially when other predisposing factors are present, is warranted about the potential risk of IFIs in ibrutinib-treated patients.

2020 - In Ph+BCR-ABL1P210+ acute lymphoblastic leukemia the e13a2 (B2A2) transcript is prevalent [Articolo su rivista]
Baccarani, M.; Iacobucci, I.; Chiaretti, S.; Foa', R.; Balasubramanian, P.; Paietta, E.; Foroni, L.; Jeromin, S.; Izzo, B.; Spinelli, O.; Varma, N.; Menif, S.; Terragna, C.; Seth, T.; Bidet, A.; Coriu, D.; Lunghi, F.; Mayer, J.; Scappini, B.; Langabeer, S.; Maier, J.; Burt, E.; Candoni, A.; Albano, F.; Luppi, M.; Zupan, I.; Lion, T.; Zadro, R.; di Raimondo, F.; Poopak, B.; Rege-Cambrin, G.; Annunziata, M.; Ayala, A.; Salinas-Viedma, V.; Ines Prado, A.; Milner, B.; Galimberti, S.; Janssen, J.; Polli, V.; Comba, L.; Borsellino, B.; Annibali, O.; Crugnola, M.; Passamonti, F.
abstract

2020 - Independent research on cancer pain management in the setting of early palliative care: A flywheel to counteract general opioid misuse and abuse [Articolo su rivista]
Bandieri, E.; Potenza, L.; Efficace, F.; Bruera, E.; Luppi, M.
abstract

The increased recognition of the high prevalence and important burden of cancer pain and the documentation of a large proportion of patients receiving inadequate analgesic treatment should have reinforced the need for evidence-based recommendations. The World health Organization (WHO) guidelines on cancer pain management—or palliative care—are traditionally based on a sequential, three-step, analgesic ladder according to pain intensity: nonopioids (paracetamol or nonsteroidal anti-inflammatory drugs) to mild pain in step I; weak opioids (eg, codeine or tramadol) to mild-moderate pain in step II; and strong opioids to moderate-severe pain in step. III. Despite the widespread use of this ladder, unrelieved pain continues to be a substantial concern in one third of patients with either solid or hematologic malignancies. The sequential WHO analgesic ladder, and in particular, the usefulness of step II opioids have been questioned but there are no universally used guidelines for the treatment of pain in patients with advanced cancer and not all guideline recommendations are evidence-based. The American Society of Clinical Oncology and the European Society of Medical Oncology have recommended the implementation of early palliative care (EPC), which is a novel model of care, consisting of delivering dedicated palliative service concurrent with active treatment as early as possible in the cancer disease trajectory. Improvement in cancer pain management is one of the several important positive effects following EPC interventions. Independent well-designed research studies on pharmacological interventions on cancer pain, especially in the EPC setting are warranted and may contribute to spur research initiatives to investigate the poorly addressed issues of pain management in non cancer patients.

2020 - Investigating the association between physicians self-efficacy regarding communication skills and risk of “burnout” [Articolo su rivista]
Messerotti, Andrea; Banchelli, Federico; Ferrari, Silvia; Barbieri, Emiliano; Bettelli, Francesca; Bandieri, Elena; Giusti, Davide; Catellani, Hillary; Borelli, Eleonora; Colaci, Elisabetta; Pioli, Valeria; Morselli, Monica; Forghieri, Fabio; Galeazzi, Gian Maria; Marasca, Roberto; Bigi, Sarah; D’Amico, Roberto; Martin, Peter; Efficace, Fabio; Luppi, Mario; Potenza, Leonardo
abstract

2020 - More on: ‘COVID-19 coagulopathy in Caucasian patients’ [Articolo su rivista]
Marietta, M.; Coluccio, V.; Luppi, M.
abstract

2020 - Npm1‐mutated myeloid neoplasms with <20% blasts: A really distinct clinico‐pathologic entity? [Articolo su rivista]
Forghieri, F.; Nasillo, V.; Paolini, A.; Bettelli, F.; Pioli, V.; Giusti, D.; Gilioli, A.; Colasante, C.; Acquaviva, G.; Riva, G.; Barozzi, P.; Maffei, R.; Potenza, L.; Marasca, R.; Fozza, C.; Tagliafico, E.; Trenti, T.; Comoli, P.; Longo, G.; Luppi, M.
abstract

Nucleophosmin (NPM1) gene mutations rarely occur in non‐acute myeloid neoplasms (MNs) with &lt;20% blasts. Among nearly 10,000 patients investigated so far, molecular analyses documented NPM1 mutations in around 2% of myelodysplastic syndrome (MDS) cases, mainly belonging to MDS with excess of blasts, and 3% of myelodysplastic/myeloproliferative neoplasm (MDS/MPN) cases, prevalently classified as chronic myelomonocytic leukemia. These uncommon malignancies are associated with an aggressive clinical course, relatively rapid progression to overt acute myeloid leukemia (AML) and poor survival outcomes, raising controversies on their classification as distinct clinico‐pathologic entities. Furthermore, fit patients with NPM1‐mutated MNs with &lt;20% blasts could benefit most from upfront intensive chemotherapy for AML rather than from moderate intensity MDS‐directed therapies, although no firm conclusion can currently be drawn on best therapeutic approaches, due to the limited available data, obtained from small and mainly retrospective series. Caution is also suggested in definitely diagnosing NPM1‐mutated MNs with blast count &lt;20%, since NPM1‐mutated AML cases frequently present dysplastic features and multilineage bone marrow cells showing abnormal cytoplasmic NPM1 protein delocalization by immunohistochemical staining, therefore belonging to NPM1‐mutated clone regardless of blast morphology. Further prospective studies are warranted to definitely assess whether NPM1 mutations may become sufficient to diagnose AML, irrespective of blast percentage.

2020 - Polymorphisms within the ARNT2 and CX3CR1 Genes Are Associated with the Risk of Developing Invasive Aspergillosis [Articolo su rivista]
Lupianez, C. B.; Martinez-Bueno, M.; Sanchez-Maldonado, J. M.; Badiola, J.; Cunha, C.; Springer, J.; Lackner, M.; Segura-Catena, J.; Canet, L. M.; Alcazar-Fuoli, L.; Lopez-Nevot, M. A.; Fianchi, L.; Aguado, J. M.; Pagano, L.; Lopez-Fernandez, E.; Alarcon-Riquelme, M.; Potenza, L.; Goncalves, S. M.; Luppi, M.; Moratalla, L.; Solano, C.; Sampedro, A.; Gonzalez-Sierra, P.; Cuenca-Estrella, M.; Lagrou, K.; Maertens, J. A.; Lass-Florl, C.; Einsele, H.; Vazquez, L.; Loeffler, J.; Rios-Tamayo, R.; Carvalho, A.; Jurado, M.; Sainz, J.
abstract

Invasive aspergillosis (IA) is a life-threatening infection that affects an increasing number of patients undergoing chemotherapy or allo-transplantation, and recent studies have shown that genetic factors contribute to disease susceptibility. In this two-stage, population-based, case-control study, we evaluated whether 7 potentially functional single nucleotide polymorphisms (SNPs) within the ARNT2 and CX3CR1 genes influence the risk of IA in high-risk hematological patients. We genotyped selected SNPs in a cohort of 500 hematological patients (103 of those had been diagnosed with proven or probable IA), and we evaluated their association with the risk of developing IA. The association of the most interesting markers of IA risk was then validated in a replication population, including 474 subjects (94 IA and 380 non-IA patients). Functional experiments were also performed to confirm the biological relevance of the most interesting markers. The meta-analysis of both populations showed that carriers of the ARNT2rs1374213G, CX3CR1rs7631529A, and CX3CR1rs9823718G alleles (where the RefSeq identifier appears as a subscript) had a significantly increased risk of developing IA according to a log-additive model (P value from the meta-analysis [PMeta] = 9.8 · 10-5, PMeta = 1.5 · 10-4, and PMeta =7.9 · 10-5, respectively). Haplotype analysis also confirmed the association of the CX3CR1 haplotype with AG CGG with an increased risk of IA (P = 4.0 · 10-4). Mechanistically, we observed that monocyte-derived macrophages (MDM) from subjects carrying the ARNTR2rs1374213G allele or the GG genotype showed a significantly impaired fungicidal activity but that MDM from carriers of the ARNT2rs1374213G and CX3CR1rs9823718G or CX3CR1rs7631529A alleles had deregulated immune responses to Aspergillus conidia. These results, together with those from expression quantitative trait locus (eQTL) data browsers showing a strong correlation of the CX3CR1rs9823718G allele with lower levels of CX3CR1 mRNA in whole peripheral blood (P = 2.46 · 10-7) and primary monocytes (P = 4.31 · 10-7), highlight the role of the ARNT2 and CX3CR1 loci in modulating and predicting IA risk and provide new insights into the host immune mechanisms involved in IA development.

2020 - Selective inhibition of PI3Kγ affects survival and proliferation of chronic lymphocytic leukemia B cells [Articolo su rivista]
Maffei, R.; Benatti, S.; Atene, C. G.; Debbia, G.; Zucchini, P.; Potenza, L.; Luppi, M.; Fiorcari, S.; Marasca, R.
abstract

the catalytic p110gamma of PI3K is implicated in the survival and proliferation of CLL cells. Our findings support the idea that CLL cells are peculiar in the attitude to sense microenvironmental signals throughout the engagement of multiple PI3Ks. PI3Kgamma inhibition by IPI-549 may directly promote CLL apoptosis, but may also interfere with signals derived from several accessory cells of stromal and immune system. Together with the reported ability of PI3Kgamma inhibition in prevention of CLL migration and adhesion, our data provide knowledge to justify further clinical development of PI3Kc inhibition in CLL cells.

2020 - The IPSS-R more accurately captures fatigue severity of newly diagnosed patients with myelodysplastic syndromes compared with the IPSS index [Articolo su rivista]
Efficace, F.; Cottone, F.; Oswald, L. B.; Cella, D.; Patriarca, A.; Niscola, P.; Breccia, M.; Platzbecker, U.; Palumbo, G. A.; Caocci, G.; Stauder, R.; Ricco, A.; Petranovic, D.; Caers, J.; Luppi, M.; Fianchi, L.; Frairia, C.; Capodanno, I.; Follini, E.; Sarlo, C.; Fazi, P.; Vignetti, M.
abstract

We aimed to compare fatigue of newly diagnosed patients with myelodysplastic syndromes (MDS) with that of the general population (GP). We also investigated the ability of the IPSS and IPSS-R to capture severity of patient-reported fatigue at diagnostic workup. A sample of 927 newly diagnosed patients with MDS was consecutively enrolled in a large international observational study and all patients completed the FACIT-Fatigue questionnaire at baseline. Fatigue was compared with that of the GP (N = 1075) and a 3-point difference in mean scores was considered as clinically meaningful. Fatigue of MDS patients was on average 4.6 points below the mean of the GP (95% CI, −5.9 to −3.2, p &lt; 0.001), reflecting clinically meaningful worse fatigue. Unlike the IPSS, the IPSS-R identified clearly distinct subgroups with regard to burden of fatigue. Mean scores differences compared with GP ranged from nonclinically relevant for very low risk (Δ = −1.8, 95% CI, −4.0 to 0.5, p = 0.119) to large clinically meaningful differences for very high-risk IPSS-R patients (Δ = −8.2, 95% CI, −10.3 to −6.2, p &lt; 0.001). At diagnostic workup, fatigue of MDS is clinically meaningful worse than that reported by the GP. Compared with the IPSS classification, the IPSS-R provides a better stratification of patients with regard to fatigue severity.

2020 - Two fatal cases of acute liver failure due to HSV-1 infection in COVID-19 patients following immunomodulatory therapies. [Articolo su rivista]
Busani, S; Bedini, A; Biagioni, E; Serio, L; Tonelli, R; Meschiari, M; Franceschini, E; Guaraldi, G; Cossarizza, A; Clini, E; Maiorana, A; Gennari, W; De Maria, N; Luppi, M; Mussini, C; Girardis, M.; Gibellini, Lara
abstract

We reported two fatal cases of acute liver failure secondary to Herpes Simplex Virus 1 infection in COVID-19 patients, following tocilizumab and corticosteroid therapy. Screening for and prompt recognition of Herpes Simplex Virus 1 reactivation in these patients, undergoing immunomodulatory treatment, may have potentially relevant clinical consequences.

2020 - Younger age at diagnosis of acute promyelocytic leukaemia is associated with better long-term cognitive functioning [Articolo su rivista]
Breccia, M.; Vignetti, M.; Annibali, O.; Cottone, F.; Luppi, M.; Borlenghi, E.; Rodeghiero, F.; Efficace, F.
abstract

2019 - Breakthrough invasive fungal diseases in acute myeloid leukemia patients receiving mould active triazole primary prophylaxis after intensive chemotherapy: An Italian consensus agreement on definitions and management [Articolo su rivista]
Girmenia, C.; Busca, A.; Candoni, A.; Cesaro, S.; Luppi, M.; Nosari, A. M.; Pagano, L.; Rossi, G.; Venditti, A.; Aversa, F.
abstract

In the attempt to establish definitions and provide shared approaches to breakthrough invasive fungal diseases (br-IFD) in acute myeloid leukemia (AML) patients submitted to intensive chemotherapy and receiving triazoles as mould active primary antifungal prophylaxis (MA-PAP), literature on br-IFD in AML patients receiving triazoles MA-PAP was reviewed and a Consensus Development Conference Project was convened. The following four candidate key-questions were generated and formed the set of questions of the present document: “definition of br-IFD,” “diagnostic strategy during MA-PAP to detect br-IFD,” “possible causes of MA-PAP failure,” “management of br-IFD.

2019 - Characterization and dynamics of specific T cells against nucleophosmin-1 (NPM1)-mutated peptides in patients with NPM1-mutated acute myeloid leukemia [Articolo su rivista]
Forghieri, Fabio; Riva, Giovanni; Lagreca, Ivana; Barozzi, Patrizia; Vallerini, Daniela; Morselli, Monica; Paolini, Ambra; Bresciani, Paola; Colaci, Elisabetta; Maccaferri, Monica; Gilioli, Andrea; Nasillo, Vincenzo; Messerotti, Andrea; Pioli, Valeria; Arletti, Laura; Giusti, Davide; Bettelli, Francesca; Celli, Melania; Donatelli, Francesca; Corradini, Giorgia; Basso, Sabrina; Gurrado, Antonella; Cellini, Monica; Trenti, Tommaso; Marasca, Roberto; Narni, Franco; Martelli, Maria Paola; Falini, Brunangelo; Potenza, Leonardo; Luppi, Mario; Comoli, Patrizia
abstract

Nucleophosmin(NPM1)-mutated protein, a leukemia-specific antigen, represents an ideal target for AML immunotherapy. We investigated the dynamics of NPM1-mutated-specific T cells on PB and BM samples, collected from 31 adult NPM1-mutated AML patients throughout the disease course, and stimulated with mixtures of 18 short and long peptides (9-18mers), deriving from the complete C-terminal of the NPM1-mutated protein. Two 9-mer peptides, namely LAVEEVSLR and AVEEVSLRK (13.9-14.9), were identified as the most immunogenic epitopes. IFNγ-producing NPM1-mutated-specific T cells were observed by ELISPOT assay after stimulation with peptides 13.9-14.9 in 43/85 (50.6%) PB and 34/80 (42.5%) BM samples. An inverse correlation between MRD kinetics and anti-leukemic specific T cells was observed. Cytokine Secretion Assays allowed to predominantly and respectively identify Effector Memory and Central Memory T cells among IFNγ-producing and IL2-producing T cells. Moreover, NPM1-mutated-specific CTLs against primary leukemic blasts or PHA-blasts pulsed with different peptide pools could be expanded ex vivo from NPM1-mutated AML patients or primed in healthy donors. We describe the spontaneous appearance and persistence of NPM1-mutated-specific T cells, which may contribute to the maintenance of long-lasting remissions. Future studies are warranted to investigate the potential role of both autologous and allogeneic adoptive immunotherapy in NPM1-mutated AML patients.

2019 - Clinical differences in sarcoidosis patients with and without lymphoma: a single-center retrospective cohort analysis. [Articolo su rivista]
Cerri, Stefania; Fontana, Matteo; Balduzzi, Sara; Potenza, Leonardo; Faverio, Paola; Luppi, Mario; Damico, Roberto; Spagnolo, Paolo; Clini, Enrico; Luppi, Fabrizio
abstract

We retrospectively reviewed the database&nbsp;of the “Center for Rare Lung Diseases” at the University Hospital of Modena to identify all subjects with a diagnosis of sarcoidosis between 1990 and 2013, with the aim to evaluate clinical, functional and serological differences related to the presence of lymphoma in sarcoidosis patients, as well as difference in survival. This study suggests the existence of clinical, radiological and serological differences in sarcoidosis with or without lymphoma syndrome. The knowledge of these differences seems important for a timely diagnosis and treatment. However, further prospective studies are required to confirm present observations.

2019 - Direct oral anticoagulants for atrial fibrillation in patients with congenital factor VII deficiency [Articolo su rivista]
Arletti, L.; Coluccio, V.; Romagnoli, E.; Luppi, M.; Marietta, M.
abstract

The management of anticoagulant therapy (OAT) in patients with factor VII (FVII) deficiency is a very challenging clinical issue, as warfarin further reduces FVII levels, thus potentially increasing bleeding risk. On the other hand, the International Normalized Ratio test is misleading in such patients, as they do not reflect the actual level of global inhibition of the coagulation system. We report here three cases of patients with a moderate FVII deficiency and receiving direct oral anticoagulants (DOAC) for prevention of cardioembolism in atrial fibrillation. Of note, two of them experienced a treatment failure while on warfarin, while DOAC treatment was not associated with thrombotic or hemorrhagic adverse events. DOAC are very attractive for the management of OAT in FVII deficient patients, because they do not require monitoring by tests affected by the inherited defect, and their mechanism of action is FVII-independent.

2019 - Effects of listening to live singing in patients undergoing hemodialysis: a randomized controlled crossover study [Articolo su rivista]
Burrai, F; Lupi, R; Luppi, M; Micheluzzi, V; Donati, G; La Manna, G; Raghavan, R
abstract

Abstract Background: Participation in music therapy is associated with improved psychological and physical indices among chronically ill patients. Listening to music during hemodialysis treatments positively affects patients’ hemodynamics, laboratory values, quality of life, and physical symptoms. The effect of live singing during hemodialysis treatments, however, has not previously been studied. Methods: A total of 24 participants with a diagnosis of end-stage kidney disease participated in the study. The vocalist was a musically trained dialysis nurse. Twelve of the patients listened to 15 min of live singing during 6 consecutive hemodialysis sessions, while the other 12 underwent standard hemodia lysis. After a washout period of 2 days, the two groups were reversed. Results: Listening to live music was associated with improvements in systolic and diastolic blood pressure, better quality of sleep, fewer cramps, and reduced anxiety/depression, pain, and itching ( p &lt; .05, all values). Conclusions: Listening to live music during hemodialysis is an effective and potentially low-cost therapy for the dialysis care team to employ during hemodialysis treatments

2019 - GIMEMA AML1310 trial of risk-adapted, MRD-directed therapy for young adults with newly diagnosed acute myeloid leukemia [Articolo su rivista]
Venditti, A.; Piciocchi, A.; Candoni, A.; Melillo, L.; Calafiore, V.; Cairoli, R.; De Fabritiis, P.; Storti, G.; Salutari, P.; Lanza, F.; Martinelli, G.; Luppi, M.; Mazza, P.; Martelli, M. P.; Cuneo, A.; Albano, F.; Fabbiano, F.; Tafuri, A.; Chierichini, A.; Tieghi, A.; Fracchiolla, N. S.; Capelli, D.; Foa, R.; Alati, C.; Sala, E. L.; Fazi, P.; Vignetti, M.; Maurillo, L.; Buccisano, F.; Del Principe, M. I.; Irno-Consalvo, M.; Ottone, T.; Lavorgna, S.; Voso, M. T.; Lo-Coco, F.; Arcese, W.; Amadori, S.
abstract

We designed a trial in which postremission therapy of young patients with de novo acute myeloid leukemia (AML) was decided combining cytogenetics/genetics and postconsolidation levels of minimal residual disease (MRD). After induction and consolidation, favorable-risk patients (FR) were to receive autologous stem cell transplant (AuSCT) and poor-risk patients (PR) allogeneic stem cell transplant (AlloSCT). Intermediate-risk patients (IR) were to receive AuSCT or AlloSCT depending on the postconsolidation levels of MRD. Three hundred sixty-one of 500 patients (72%) achieved a complete remission, 342/361 completed the consolidation phase and were treatment allocated: 165 (48%) to AlloSCT (122 PR, 43 IR MRD-positive) plus 23 rescued after salvage therapy, for a total of 188 candidates; 150 (44%) to AuSCT (115 FR, 35 IR MRD-negative) plus 27 IR patients (8%) with no leukemia-associated phenotype, for a total of 177 candidates. Overall, 110/177 (62%) and 130/188 (71%) AuSCT or AlloSCT candidates received it, respectively. Two-year overall (OS) and disease-free survival (DFS) of the whole series was 56% and 54%, respectively. Two-year OS and DFS were 74% and 61% in the FR category, 42% and 45% in the PR category, 79% and 61% in the IR MRD-negative category, and 70% and 67% in the IR MRD-positive category. In conclusion, AuSCT may still have a role in FR and IR MRD-negative categories. In the IR MRD-positive category, AlloSCT prolongs OS and DFS to equal those of the FR category. Using all the available sources of stem cells, AlloSCT was delivered to 71% of the candidates

2019 - Health-related quality of life, symptom burden, and comorbidity in long-term survivors of acute promyelocytic leukemia [Articolo su rivista]
Efficace, Fabio; Breccia, Massimo; Avvisati, Giuseppe; Cottone, Francesco; Intermesoli, Tamara; Borlenghi, Erika; Carluccio, Paola; Rodeghiero, Francesco; Fabbiano, Francesco; Luppi, Mario; Romani, Claudio; Sborgia, Marco; D’Ardia, Stefano; Nobile, Francesco; Cantore, Nicola; Crugnola, Monica; Nadali, Gianpaolo; Vignetti, Marco; Amadori, Sergio; Lo Coco, Francesco
abstract

The objective of this study was to investigate health-related quality of life (HRQOL), symptom burden, and comorbidity profile in long-term acute promyelocytic leukemia (APL) survivors treated with standard chemotherapy. Overall, 307 long-term APL survivors were invited to participate. HRQOL was assessed with the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and compared with that of age and sex-matched controls from the general population. Symptom burden was assessed with the MD Anderson Symptom Inventory (MDASI) questionnaire and comorbidity profile was also investigated. Median follow-up time since diagnosis was 14.3 years (interquartile range: 11.1–16.9 years). APL survivors had a statistically and clinically meaningful worse score for the role physical scale of the SF-36 (−9.5; 95% CI, −15.7 to −3.2, P = 0.003) than their peers in the general population. Fatigue was reported as moderate to severe by 29% of patients and 84.4% reported at least one comorbidity. Prevalence of comorbidity in APL survivors was higher than that reported by the general population. Also, marked variations were found in the HRQOL profile by number of comorbidities. Even many years after treatment ends, APL survivors treated with standard chemotherapy do not fully recover as they report HRQOL limitations and a substantial burden of symptoms.

2019 - Mindfulness-Based stress reduction in early palliative care for people with metastatic cancer: A mixed-method study [Articolo su rivista]
Poletti, S.; Razzini, G.; Ferrari, R.; Ricchieri, M. P.; Spedicato, G. A.; Pasqualini, A.; Buzzega, C.; Artioli, F.; Petropulacos, K.; Luppi, M.; Bandieri, E.
abstract

Objectives: To explore the impact of a Mindfulness-Based Stress Reduction (MBSR) intervention for people with metastatic cancer integrated in Early Palliative Care (EPC). Design: Mixed-method study. Settings/Location: EPC Service integrated with Oncology Unit, Carpi General Hospital, Italy from January to October 2017. The MBSR intervention took place inside the hospital. Subjects: Study participation was offered to 25 consecutive people referred to the EPC service. Inclusion criteria: people with metastatic cancer between 18 and 75 years old; informed consent. Exclusion criteria: Performance Status &lt;60% according to Karnofsky scale; active psychiatric disorder. 20 patients were included in the study. Intervention: The adapted program consists of 8 meetings for 2.5 h once a week, a 4.5 h session between the 6th and 7th weeks and 0.5 h home practice daily. The following mindfulness practices were included during the training: formal sitting meditation, body scan, light yoga, walking meditation, and Aikido exercises. Participants were provided with materials for home practice. A qualified MBSR instructor conducted the program. Sessions were attended by a clinical psychologist and a physician trained in meditation, together with the palliative nurse as facilitators. Outcome Measures: Feasibility and acceptability were assessed on 16 participants. In addition, pre-post measures of cancer pain and mood state were collected. Semi-structured, in-depth interviews were conducted on a subset of 8 participants at the end of the study and analysed using the Interpretative-Phenomenological approach. Results: MBSR attendance to meetings and adherence to home practice were 75%. MBSR intervention helped participants to develop an accepting attitude in respect to metastatic cancer disease helping them to face anxiety and cancer pain. MBSR improves self-regulation of mood state engendering feelings of compassion MBSR program supports participants in questioning and reconnecting with their values and spiritual beliefs. Conclusions: A Mindfulness intervention integrated into EPC setting is feasible, well accepted and could help metastatic cancer patients to control cancer pain together with an opportunity of emotional and spiritual relief.

2019 - Overexpression of CD49d in trisomy 12 chronic lymphocytic leukemia patients is mediated by IRF4 through induction of IKAROS [Articolo su rivista]
Fiorcari, Stefania; Benatti, Stefania; Zucchetto, Antonella; Zucchini, Patrizia; Gattei, Valter; Luppi, Mario; Marasca, Roberto; Maffei, Rossana
abstract

2019 - Pomalidomide, bortezomib, and dexamethasone for patients with relapsed or refractory multiple myeloma previously treated with lenalidomide (OPTIMISMM): a randomised, open-label, phase 3 trial [Articolo su rivista]
Richardson, P. G.; Oriol, A.; Beksac, M.; Liberati, A. M.; Galli, M.; Schjesvold, F.; Lindsay, J.; Weisel, K.; White, D.; Facon, T.; San Miguel, J.; Sunami, K.; O'Gorman, P.; Sonneveld, P.; Robak, P.; Semochkin, S.; Schey, S.; Yu, X.; Doerr, T.; Bensmaine, A.; Biyukov, T.; Peluso, T.; Zaki, M.; Anderson, K.; Dimopoulos, M.; Abildgaard, N.; Adler, H.; Altuntas, F.; Akay, O. M.; Amin, B.; Anagnostopoulos, A.; Anderson, L.; Anttila, P.; Araujo, C.; Arce-Lara, C.; Aydin, Y.; Basu, S.; Battini, R.; Beeker, T.; Benboubker, L.; Ben-Yehuda, D.; Blade, J.; Blau, I. W.; Boccia, R.; Burke, L.; Byeff, P.; Cascavilla, N.; Cavo, M.; Chantry, A.; Charles, Y.; Chaudhry, A.; Corso, A.; Coyne, M.; De Arriba, F.; Delimpasi, S.; Desjardins, P.; Dhakal, B.; Di Bartolomeo, P.; Di Raimondo, F.; Durig, J.; Engelhardt, M.; Escoffre-Barbe, M.; Esteves, G.; Flogegard, M.; Gabrail, N.; Gamberi, B.; Garrison, M.; Gay, J.; Gisslinger, H.; Goldschmidt, H.; Goncalves, C.; Gressot, L.; Grosicki, S.; Hanna, W.; Hayden, P.; Henriques Bernardo, M. M.; Hermann, R.; Holden, V.; Honkalehto, K.; Huben, M.; Huffman, J.; Hunter, H.; Hus, M.; Jagasia, M.; Jagganath, S.; Janakiram, M.; Jaiyesimi, I.; Jenner, M.; Joao, C.; Johnson, P.; Jurcyszyn, A.; Kalayoglu Besisik, S.; Kambhampati, S.; Kanate, A.; Karadogan, I.; Khojasteh, A.; Kirkel, D.; Komarnicki, M.; Krauth, M. -T.; Kuriakose, P.; Larocca, A.; Lauri, B.; Leleu, X.; Lucio, P.; Luppi, M.; Mangiacavalli, S.; Mariette, C.; Matsue, K.; Mellqvist, U. -H.; Mendeleeva, L.; Meshad, M.; Miller, C.; Mohrbacher, A.; Moreau, P.; Morelli, A. M.; Muldur, E.; Naassan, A.; Nahi, H.; Nair, R.; O'Dwyer, M.; Ongoren Aydin, S.; Openshaw, T.; O'Rourke, T.; Osswald, M.; Overton, L.; Pati, A.; Pavic, M.; Pegourie, B.; Pehlivan, M.; Pierola, A. A.; Plesner, T.; Pluta, A.; Rabin, N.; Ramasamy, K.; Rambaldi, A.; Rodriguez, P.; Rollig, C.; Rosenblatt, J.; Rosenbluth, J.; Salomo, M.; Samoylova, O.; Sastre Moral, J.; Sati, H.; Selleri, C.; Shafeek, S.; Shinagawa, A.; Sleckman, B.; Smith, C.; Sonmez, M.; Stone, C.; Streetly, M.; Suzuki, K.; Taetle, R.; Tafuri, A.; Takezako, N.; Teke, H. U.; Vapaatalo, M.; Vassilopoulos, G.; Verma, A.; Vidito, S.; Viterbo, L.; Vural, F.; Wang, X. S.; Yagci, M.; Yee, A.
abstract

Background: As lenalidomide becomes increasingly established for upfront treatment of multiple myeloma, patients refractory to this drug represent a population with an unmet need. The combination of pomalidomide, bortezomib, and dexamethasone has shown promising results in phase 1/2 trials of patients with relapsed or refractory multiple myeloma. We aimed to assess the efficacy and safety of this triplet regimen in patients with relapsed or refractory multiple myeloma who previously received lenalidomide. Methods: We did a randomised, open-label, phase 3 trial at 133 hospitals and research centres in 21 countries. We enrolled patients (aged ≥18 years)with a diagnosis of multiple myeloma and measurable disease, an Eastern Cooperative Oncology Group performance status of 0–2, who received one to three previous regimens, including a lenalidomide-containing regimen for at least two consecutive cycles. We randomly assigned patients (1:1)to bortezomib and dexamethasone with or without pomalidomide using a permutated blocked design in blocks of four, stratified according to age, number of previous regimens, and concentration of β2 microglobulin at screening. Bortezomib (1·3 mg/m2)was administered intravenously until protocol amendment 1 then either intravenously or subcutaneously on days 1, 4, 8, and 11 for the first eight cycles and subsequently on days 1 and 8. Dexamethasone (20 mg [10 mg if age &gt;75 years])was administered orally on the same days as bortezomib and the day after. Patients allocated pomalidomide received 4 mg orally on days 1–14. Treatment cycles were every 21 days. The primary endpoint was progression-free survival in the intention-to-treat population, as assessed by an independent review committee. Safety was assessed in all patients who received at least one dose of study medication. This trial is registered at ClinicalTrials.gov, number NCT01734928; patients are no longer being enrolled. Findings: Between Jan 7, 2013, and May 15, 2017, 559 patients were enrolled. 281 patients were assigned pomalidomide, bortezomib, and dexamethasone and 278 were allocated bortezomib and dexamethasone. Median follow-up was 15·9 months (IQR 9·9–21·7). Pomalidomide, bortezomib, and dexamethasone significantly improved progression-free survival compared with bortezomib and dexamethasone (median 11·20 months [95% CI 9·66–13·73]vs 7·10 months [5·88–8·48]; hazard ratio 0·61, 95% CI 0·49–0·77; p&lt;0·0001). 278 patients received at least one dose of pomalidomide, bortezomib, and dexamethasone and 270 patients received at least one dose of bortezomib and dexamethasone, and these patients were included in safety assessments. The most common grade 3 or 4 treatment-emergent adverse events were neutropenia (116 [42%]of 278 patients vs 23 [9%]of 270 patients; nine [3%]vs no patients had febrile neutropenia), infections (86 [31%]vs 48 [18%]), and thrombocytopenia (76 [27%]vs 79 [29%]). Serious adverse events were reported in 159 (57%)of 278 patients versus 114 (42%)of 270 patients. Eight deaths were related to treatment; six (2%)were recorded in patients who received pomalidomide, bortezomib, and dexamethasone (pneumonia [n=2], unknown cause [n=2], cardiac arrest [n=1], cardiorespiratory arrest [n=1])and two (1%)were reported in patients who received bortezomib and dexamethasone (pneumonia [n=1], hepatic encephalopathy [n=1]). Interpretation: Patients with relapsed or refractory multiple myeloma who previously received lenalidomide had significantly improved progression-free survival when treated with pomalidomide, bortezomib, and dexamethasone compared with bortezomib and dexamethasone. Adverse events accorded with the individual profiles of pomalidomide, bortezomib, and dexamethasone. This study supports use of pomalidomide, bortezomib, and dexamethasone as a treatment option in patients with relapsed or refractory multiple myeloma who previously received lenalidomide. Funding: Celgene.

2019 - Rhodotorula infection in haematological patient: risk factors and outcome [Articolo su rivista]
Potenza, Leonardo; Chitasombat, Maria N; Klimko, Nikolay; Bettelli, Francesca; Dragonetti, Giulia; Del Principe, Maria Ilaria; Nucci, Marcio; Busca, Alessandro; Fracchiolla, Nicola; Sciumè, Mariarita; Spolzino, Angelica; Delia, Mario; Mancini, Valentina; Nadali, Gian Paolo; Dargenio, Michela; Shadrivova, Olga; Bacchelli, Federico; Aversa, Franco; Sanguinetti, Maurizio; Luppi, Mario; Kontoyiannis, Dimitrios P; Pagano, Livio
abstract

Rhodotorula spp are uncommon yeasts able to cause infections with high mortality rates. Rhodotorula infections have been associated with the presence of central venous catheter (CVC), immunosuppression, exposure to antifungals and the presence of either solid or hematologic malignancies. However, in this latter setting, only a few cases have so far been reported.

2019 - T-cell large granular lymphocyte leukemia in solid organ transplant recipients: case series and review of the literature [Articolo su rivista]
Alfano, G.; Fontana, F.; Colaci, E.; Mori, G.; Cerami, C.; Messerotti, A.; Potenza, L.; Luppi, M.; Cappelli, G.
abstract

T-cell large granular lymphocyte (T-LGL) leukemia is a rare clonal proliferation of cytotoxic lymphocytes rarely described in solid organ transplant (SOT). We reviewed records from 656 kidney transplant recipients in follow-up at our Center from January 1998 to July 2017. In addition, we researched, through PubMed, further&nbsp;reports of T-LGL leukemia in SOT from March 1981 to December 2017. We identified six cases of T-LGL leukemia in our cohort of patients and 10 in the literature. This lymphoproliferative disorder was detected in one combined liver–kidney, one liver and 14–kidney transplant recipients. Median age at presentation was 46.5&nbsp;years (IQR 39.2–56.9). The disease developed after a median age of 10&nbsp;years (IQR 4.9–12) from transplantation. Anemia was the most common presentation (62.5%) followed by lymphocytosis (43.7%) and thrombocytopenia (31.2%). Splenomegaly was reported in 43.7% of the patients. Eight patients (50%) who experienced severe symptoms were treated with non-specific immunosuppressive agents. Six of them (75%) had a good outcome, whereas two (25%) remained red blood cell transfusion dependent. No cases progressed to aggressive T-LGL leukemia or died of cancer at the end of follow-up. These results suggest that T-LGL leukemia is a rare but potentially disruptive hematological disorder in the post-transplant period.

2018 - A typical atypical chronic myeloid leukemia [Articolo su rivista]
Gilioli, Andrea; Paolini, Ambra; Bonacorsi, Goretta; Luppi, Mario
abstract

Key Clinical MessageIn the context of leukocytosis due to increased number of neutrophils and their precursors, with significant dysgranulopoiesis and no or minimal basophilia and no or minimal monocytosis, the typical feature of "clumped" chromatin, in irregularly coarse compacted nuclei, should lead to suspect the diagnosis of atypical chronic myeloid leukemia.

2018 - Angiopoietin-2 acts as a survival factor for chronic lymphocytic leukemia B cells throughout Tie-2 receptor engagement [Articolo su rivista]
Maffei, Rossana; Fiorcari, Stefania; Martinelli, Silvia; Guarnotta, Carla; Benatti, Stefania; Belmonte, Beatrice; Potenza, Leonardo; Luppi, Mario; Marasca, Roberto
abstract

we demonstrated that: (i) CLL cells expressed Tie-2 receptor both in peripheral blood and in lymph nodes; (ii) Ang2 may interact with Tie-2 in CLL mediating a survival signal throughout PI3K-AKT signalling, and (iii) the interruption of Ang2/Tie-2 signalling may be effective in CLL.

2018 - Breast Location for De Novo Extramedullary Myeloid Sarcoma [Articolo su rivista]
Tazzioli, Giovanni; Palma, Enza; Anna, Gambini; Messerotti, Andrea; Potenza, Leonardo; Luppi, Mario; Drago, Antonella; Grazia Amorico, Maria; Barbolini, Monica; Ficarra, Guido; Piacentini, Federico
abstract

Background: Myeloid Sarcoma (MS) is a rare hematologic cancer, which can occur as a breast mass to be distinguished from other non-hematopoietic tumors. Case Presentation: This report describe the unusual clinical history of a young woman diagnosed with MS. Radiotherapy/surgery alone may be inadequate, while chemotherapy and hematopoietic stem cell transplantation demonstrated to improve the prognosis for the isolated extramedullary localization. Conclusion: Performing a needle biopsy in order to exclude the diagnosis of a primitive breast disease is irreplaceable.

2018 - Dual inhibition of PI3K/mTOR signaling in chemoresistant AMLprimary cells. [Articolo su rivista]
Bertacchini, Jessika; Frasson, Chiara; Chiarini, Francesca; D'Avella, Daniele; Accordi, Benedetta; Anselmi, Laura; Barozzi, Patrizia; Foghieri, Fabio; Luppi, Mario; Martelli, Alberto M.; Basso, Giuseppe; Najmaldin, Saki; Khosravi, Abbas; Rahim, Fakher; Marmiroli, Sandra
abstract

A main cause of treatment failure for AML patients is resistance to chemotherapy. Survival of AML cells may depend on mechanisms that elude conventional drugs action and/or on the presence of leukemia initiating cells at diagnosis, and their persistence after therapy. MDR1 gene is an ATP-dependent drug efflux pump known to be a risk factor for the emergence of resistance, when combined to unstable cytogenetic profile of AML patients. In the present study, we analyzed the sensitivity to conventional chemotherapeutic drugs of 26 samples of primary blasts collected from AML patients at diagnosis. Detection of cell viability and apoptosis allowed to identify two group of samples, one resistant and one sensitive to in vitro treatment. The cells were then analyzed for the presence and the activity of P-glycoprotein. A comparative analysis showed that resistant samples exhibited a high level of MDR1 mRNA as well as of P-glycoprotein content and activity. Moreover, they also displayed high PI3K signaling. Therefore, we checked whether the association with signaling inhibitors might resensitize resistant samples to chemo-drugs. The combination showed a very potent cytotoxic effect, possibly through down modulation of MDR1, which was maintained also when primary blasts were co-cultured with human stromal cells. Remarkably, dual PI3K/mTOR inactivation was cytotoxic also to leukemia initiating cells. All together, our findings indicate that signaling activation profiling associated to gene expression can be very useful to stratify patients and improve therapy.

2018 - Effectiveness of originator (Neupogen) and biosimilar (Zarzio) filgrastim in autologous peripheral blood stem cell mobilization in adults with acute myeloid leukemia: a single-center retrospective study [Articolo su rivista]
Nasillo, Vincenzo; Paolini, Ambra; Riva, Giovanni; Morselli, Monica; Potenza, Leonardo; Coluccio, Valeria; Maccaferri, Monica; Colaci, Elisabetta; Fantuzzi, Valeria; Messerotti, Andrea; Arletti, Laura; Pioli, Valeria; Lugli, Elisabetta; Gilioli, Andrea; Quadrelli, Chiara; Zucchini, Patrizia; Vallerini, Daniela; Lagreca, Ivana; Barozzi, Patrizia; Cuoghi, Angela; Bresciani, Paola; Marasca, Roberto; Mariano, Maria Teresa; Ceccherelli, Giovanni; Comoli, Patrizia; Campioli, Daniele; Trenti, Tommaso; Narni, Franco; Luppi, Mario; Forghieri, Fabio
abstract

n.d.

2018 - Idelalisib impairs T-cell-mediated immunity in chronic lymphocytic leukemia [Articolo su rivista]
Martinelli, Silvia; Maffei, Rossana; Fiorcari, Stefania; Quadrelli, Chiara; Zucchini, Patrizia; Benatti, Stefania; Potenza, Leonardo; Luppi, Mario; Marasca, Roberto
abstract

n.d.

2018 - Immunophenotypic Profile and Clinical Outcome of Monoclonal B-cell Lymphocytosis in Kidney Transplantation [Articolo su rivista]
Alfano, G; Fontana, F; Colaci, E; Franceschini, E; Ligabue, G; Messerotti, A; Bettelli, Francesca; Grottola, A; Gennari, W; Potenza, L; Guaraldi, G; Mussini, C; Luppi, M; Cappelli, G
abstract

Monoclonal B-cell lymphocytosis (MBL) is a lymphoproliferative disorder characterized by clonal expansion of a B-cell population in peripheral blood of otherwise healthy subjects. MBL is divided into CLL (chronic lymphocytic leukemia)-like, atypical CLL-like and non-CLL MBL. The aim of this study was to evaluate immunophenotypic characteristics and clinical outcomes of MBL in kidney transplant (KT) recipients. We retrospectively evaluated 593 kidney transplant (KT) recipients in follow-up at our center. Among them, 157 patients underwent peripheral blood flow-cytometry for different clinical indications. A 6-color panel flow-cytometry was used to diagnose MBL. MBL was detected in 5 of 157 KT recipients. Immunophenotypic characterization of MBL showed four cases of non-CLL MBL and one case of CLL-like MBL. At presentation, median age was 65 years (range 61-73). After a median follow-up of 3.1 years (95%CI; 1.1-5) from diagnosis, patients did not progress either to CLL or lymphoma. The disorder did not increase the risk of malignancy, severe infections, graft loss and mortality among our KT recipients. Surprisingly, all cases were also affected by concomitant monoclonal gammopathy of undetermined significance, which did not progress to multiple myeloma during follow-up. In conclusion, our data suggest that MBL is an age-related disorder, with non-CLL MBL being the most common subtype among KT recipients. This article is protected by copyright. All rights reserved.

2018 - Increased SHISA3 expression characterizes chronic lymphocytic leukemia patients sensitive to lenalidomide [Articolo su rivista]
Maffei, Rossana; Fiorcari, Stefania; Martinelli, Silvia; Benatti, Stefania; Bulgarelli, Jenny; Rizzotto, Lara; Debbia, Giulia; Santachiara, Rita; Rigolin, Gian Matteo; Forconi, Francesco; Rossi, Davide; Laurenti, Luca; Palumbo, Giuseppe A.; Vallisa, Daniele; Cuneo, Antonio; Gaidano, Gianluca; Luppi, Mario; Marasca, Roberto
abstract

Lenalidomide is a therapeutically effective drug in chronic lymphocytic leukemia (CLL). Twenty-seven CLL patients were treated with lenalidomide in a phase II clinical trial. Ten patients were grouped as responders (R) and 6 as nonresponders (NR). We evaluated T lymphocytes, NK, monocytes and dendritic cells at baseline and after treatment. A gene expression analysis was performed on 16 CLL samples collected before treatment. The levels of immune cells or immune-related cytokines are not different between R and NR patients. However, CLL patients sensitive to lenalidomide clearly show a peculiar gene expression profile in leukemic cells. The most up-regulated gene (fold change =  +23 in R vs. NR) is Wnt inhibitor SHISA homolog 3 (SHISA3). SHISA3highCLL are characterized by a restrained activation of Wnt signaling and sensibility to lenalidomide-induced apoptosis. In conclusion, SHISA3 is a candidate gene for the identification of CLL patients who will benefit of lenalidomide treatment as single agent.

2018 - Is there a role for intervention radiology for the treatment of lower limb deep vein thrombosis in the era of direct oral anticoagulants? A comprehensive review [Articolo su rivista]
Marietta, Marco; Romagnoli, Elisa; Cosmi, Benilde; Coluccio, Valeria; Luppi, Mario
abstract

Despite recent advances in the treatment of Deep Vein Thrombosis (DVT) provided by Direct Oral Anticoagulants (DOAC), a substantial proportion of lower limb DVT patients will develop some degree of post-thrombotic syndrome (PTS) within 2 years. Systemic thrombolysis, although effective in reducing the risk of PTS and leg ulceration, is associated with a high risk of major bleeding, making it unsuitable for the vast majority of patients. A local approach, aimed at delivering the fibrinolytic drug directly into, or near to, the thrombus surface, is attractive because of the possibility of lowering of the administered drug dose, thus reducing the bleeding risks. However, even after the recent publication of the ATTRACT trial, only weak evidence is available about the efficacy and safety of Catheter Directed Thrombolysis (CDT), either alone (pharmacological technique) or in combination with additional endovascular approaches (pharmacomechanical technique, PMT) including percutaneous mechanical thrombectomy, angioplasty with or without stenting and ultrasound-assisted CDT. The present review is aimed at providing the physicians with a comprehensive evaluation of the current evidence about this relevant topic, in order to build a reliable conceptual framework for a more appropriate use of this resource.

2018 - Lymphoproliferative syndromes associated with human herpesvirus-6A and human herpesvirus-6B [Articolo su rivista]
Eliassen, Eva; Krueger, Gerhard; Luppi, Mario; Ablashi, Dharam
abstract

Human herpesvirus 6A and 6B (HHV-6A and HHV-6B) have been noted since their discovery for their T-lymphotropism. Although it has proven difficult to determine the extent to which HHV-6A and HHV-6B are involved in the pathogenesis of many diseases, evidence suggests that primary infection and reactivation of both viruses may induce or contribute to the progression of several lymphoproliferative disorders, ranging from benign to malignant and including infectious mononucleosis-like illness, drug induced hypersensitivity syndrome/drug reaction with eosinophilia and systemic symptoms (DIHS/DRESS), and nodular sclerosis Hodgkin's lymphoma. Herein, we discuss the conditions associated with the lymphoproliferative capacity of HHV-6, as well as the potential mechanisms behind them. Continued exploration on this topic may add to our understanding of the interactions between HHV-6 and the immune system and may open the doors to more accurate diagnosis and treatment of certain lymphoproliferative disorders.

2018 - Minimal/Measurable Residual Disease Monitoring in NPM1-Mutated Acute Myeloid Leukemia: A Clinical Viewpoint and Perspectives [Articolo su rivista]
Forghieri, Fabio; Comoli, Patrizia; Marasca, Roberto; Potenza, Leonardo; Luppi, Mario
abstract

Acute myeloid leukemia (AML) with NPM1 gene mutations is currently recognized as a distinct entity, due to its unique biological and clinical features. We summarize here the results of published studies investigating the clinical application of minimal/measurable residual disease (MRD) in patients with NPM1-mutated AML, receiving either intensive chemotherapy or hematopoietic stem cell transplantation. Several clinical trials have so far demonstrated a significant independent prognostic impact of molecular MRD monitoring in NPM1-mutated AML and, accordingly, the Consensus Document from the European Leukemia Net MRD Working Party has recently recommended that NPM1-mutated AML patients have MRD assessment at informative clinical timepoints during treatment and follow-up. However, several controversies remain, mainly with regard to the most clinically significant timepoints and the MRD thresholds to be considered, but also with respect to the optimal source to be analyzed, namely bone marrow or peripheral blood samples, and the correlation of MRD with other known prognostic indicators. Moreover, we discuss potential advantages, as well as drawbacks, of newer molecular technologies such as digital droplet PCR and next-generation sequencing in comparison to conventional RQ-PCR to quantify NPM1-mutated MRD. In conclusion, further prospective clinical trials are warranted to standardize MRD monitoring strategies and to optimize MRD-guided therapeutic interventions in NPM1-mutated AML patients.

2018 - Non-replacement therapy for haemophilia treatment: Fetching the east by the west [Articolo su rivista]
Marietta, Marco; Luppi, Mario
abstract

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2018 - Novel agents for acute myeloid leukemia [Articolo su rivista]
Luppi, Mario; Fabbiano, Francesco; Visani, Giuseppe; Martinelli, Giovanni; Venditti, Adriano
abstract

Acute myeloid leukemia (AML) is a complex hematological disease characterized by genetic and clinical heterogeneity. Recent advances in the understanding of AML pathogenesis have paved the way for the development of new agents targeting specific molecules or mechanisms that contribute to finally move beyond the current standard of care, which is “3 + 7” regimen. In particular, new therapeutic options such as targeted therapies (midostaurin and enasidenib), monoclonal antibodies (gemtuzumab ozogamicin), and a novel liposomal formulation of cytarabine and daunorubicin (CPX-351) have been recently approved, and will be soon available for the treatment of adult patients with AML. In this review, we will present and describe these recently approved drugs as well as selected novel agents against AML that are currently under investigation, and show the most promising results as monotherapy or in combination with chemotherapy. The selection of these emerging treatments is based on the authors’ opinion.

2018 - O blood type is a risk factor for upper gastrointestinal bleeding [Articolo su rivista]
Franchini, M.; Togliani, T.; Turdo, R.; Lucchini, G.; Bonfanti, C.; Giacomini, I.; Luppi, M.; Pilati, S.
abstract

2018 - Patient-reported outcomes enhance the survival prediction of traditional disease risk classifications: An international study in patients with myelodysplastic syndromes [Articolo su rivista]
Efficace, Fabio; Cottone, Francesco; Abel, Gregory; Niscola, Pasquale; Gaidano, Gianluca; Bonnetain, Franck; Anota, Amelie; Caocci, Giovanni; Cronin, Angel; Fianchi, Luana; Breccia, Massimo; Stauder, Reinhard; Platzbecker, Uwe; Palumbo, Giuseppe A; Luppi, Mario; Invernizzi, Rosangela; Bergamaschi, Micaela; Borin, Lorenza; Di Tucci, Anna Angela; Zhang, Huiyong; Sprangers, Mirjam; Vignetti, Marco; Mandelli, Franco
abstract

Current prognostic systems for myelodysplastic syndromes (MDS) are based on clinical, pathologic, and laboratory indicators. The objective of the current study was to develop a new patient-centered prognostic index for patients with advanced MDS by including self-reported fatigue severity into a well-established clinical risk classification: the International Prognostic Scoring System (IPSS).

2017 - Anticancer drug-loaded quantum dots engineered polymeric nanoparticles: Diagnosis/therapy combined approach [Articolo su rivista]
Belletti, Daniela; Riva, Giovanni; Luppi, Mario; Tosi, Giovanni; Forni, Flavio; Vandelli, Maria Angela; Ruozi, Barbara; Pederzoli, Francesca
abstract

Primary Effusion Lymphoma (PEL) is an HHV-8-related non Hodgkin lymphoma localized in body cavities (as pleural, peritoneal and pericardial) presenting lymphomatous effusion that, until now, lack of an effective therapy. Curcumin was reported to display pro-apoptotic effect via the inhibition of the JAK/STAT pathway, that is overexpressed in PEL cells, as consequence of virus infection. The administration of curcumin is severely restricted by its physicochemical properties, mainly its low solubility in biological fluid and consequently low bioavailability. Encapsulation into biocompatible and biodegradable PLGA nanoparticles (NPs) could be a strategy to overcome biological limits of curcumin, offering a valuable step forward for its clinical application. In this study we described single-emulsion process for curcumin loading into NPs (encapsulation efficiency about 35%). We applied a post-formulation strategy (NHS/EDC reaction) to decorate the surface of the curcumin-loaded NPs with quantum dots (QDs) as imaging agents (QDs-NPs-Cur, 24pmol of QDs per 100mg of NPs) obtaining tools useful for possible application in theranostic approach. Bifunctionalized NPs were tested in vitro on two PEL's cell line (BCBL-1 and HBL-6). The efficacy of the treatment was evaluated by cytofluorimetric assay by measuring both cell viability and cell density. We found that the NPs significantly improve the cellular effect of curcumin (respect to free drug). Moreover, by means of confocal microscopy, both the localization of bifunctional NPs and of the released drug were easily detectable. Thus, we conclude that the delivery of curcumin using bifunctional traceable NPs is a promising future approach for the diagnosis and the treatment of PEL.

2017 - BCR-ABL-specific T-cell therapy in Ph+ ALL patients on tyrosine-kinase inhibitors [Articolo su rivista]
Comoli, Patrizia; Basso, Sabrina; Riva, Giovanni; Barozzi, Patrizia; Guido, Ilaria; Gurrado, Antonella; Quartuccio, Giuseppe; Rubert, Laura; Lagreca, Ivana; Vallerini, Daniela; Forghieri, Fabio; Morselli, Monica; Bresciani, Paola; Cuoghi, Angela; Paolini, Ambra; Colaci, Elisabetta; Marasca, Roberto; Cuneo, Antonio; Iughetti, Lorenzo; Trenti, Tommaso; Narni, Franco; Foà, Robin; Zecca, Marco; Luppi, Mario; Potenza, Leonardo
abstract

Although the emergence of bone marrow (BM)-resident (p190)BCR-ABL-specific T lymphocytes has been correlated with hematologic and cytogenetic remissions in patients with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL) undergoing maintenance tyrosine-kinase inhibitor treatment, little is known about the possibility of culturing these cells ex vivo and using them in T-cell therapy strategies. We investigated the feasibility of expanding/priming (p190)BCR-ABL-specific T cells in vitro by stimulation with dendritic cells pulsed with (p190)BCR-ABL peptides derived from the BCR-ABL junctional region and alternative splicing, and of adoptively administering them to patients with relapsed disease. We report on the feasibility of producing clinical-grade BCR-ABL-specific cytotoxic T lymphocytes (CTLs), endowed with antileukemia activity, from Ph(+) ALL patients and healthy donors. We treated 3 patients with Ph(+) ALL with autologous or allogeneic (p190)BCR-ABL-specific CTLs. No postinfusion toxicity was observed, except for a grade II skin graft-versus-host disease in the patient treated for hematologic relapse. All patients achieved a molecular or hematologic complete remission (CR) after T-cell therapy, upon emergence of (p190)BCR-ABL-specific T cells in the BM. Our results show that (p190)BCR-ABL-specific CTLs are capable of controlling treatment-refractory Ph(+) ALL in vivo, and support the development of adoptive immunotherapeutic approaches with BCR-ABL CTLs in Ph(+) ALL.

2017 - Brucellosis in Chile: Description of a series of 13 cases [Articolo su rivista]
Olivares, R.; Vidal, P.; Sotomayor, C.; Norambuena, M.; Luppi, M.; Silva, F.; Cifuentes, M.
abstract

Introduction: Brucellosis is a zoonosis caused by Brucella spp. It may be acquired by consuming unpasteurized dairy products. Brucellosis has a low incidence in Chile, thus, we have a scarce data. Aim: To report and to characterize the first series of clinical cases of adult patients diagnosed with brucellosis in Chile. Methods: We describe a series of 13 clinical cases in patients diagnosed between 2000 and 2016 in three different centers in the Metropolitan Region, Chile. A retrospective analysis was performed on clinical presentation, laboratory, antibiotic treatment, morbidity and mortality. Results: The mean age was 50 years old. Eight cases had a record of consumption of unpasteurized dairy products. The most frequently reported complaints were fever. The most frequent focal point involved was the spine. Only one patient had a positive blood culture, while the diagnosis was made using serological techniques in the other part of the group. The most indicated antibiotic regimens were doxycycline-rifampicin and doxycycline-gentamicin. The hospital stay was 20 days approximately as an average. Clinical cure was achieved in all cases. Conclusions: Brucellosis is an infrequent zoonosis in Chile, and it produces a nonspecific clinical picture, so it is necessary to have high suspicion to make the diagnosis based in the antecedent of consumption of unpasteurized dairy or raw meat.

2017 - Clinical Utility of Epstein-Barr Virus Viral Load Monitoring and Risk Factors for Posttransplant Lymphoproliferative Disorders After Kidney Transplantation [Articolo su rivista]
Franceschini, Erica; Plessi, Jessica; Zona, Stefano; Santoro, Antonella; Digaetano, Margherita; Fontana, Francesco; Alfano, Gaetano; Guaraldi, Giovanni; Comoli, Patrizia; Facchini, Francesca; Potenza, Leonardo; Gennari, William; Codeluppi, Mauro; Luppi, Mario; Cappelli, Gianni; Gyssens, Inge C.; Mussini, Cristina
abstract

Background. Posttransplant lymphoproliferative disease (PTLD) is an important cause of morbidity and mortality in solid organ transplants. Epstein Barr virus (EBV) plays a major role in PTLD development. Guidelines recommend EBV viral load (VL) monitoring in high-risk populations in the first year. Methods. Retrospective observational study in all adult patients who had at least 1 EBV-VL performed in the postkidney transplant (KT) period from January 2005 to December 2014 at the Policlinico Modena Hospital. We compared patients with negative EBV-DNA to patients with positive EBV-DNA and we described PTLD developed in the study period. Results.One hundred ninety (36.3%) KT patients of 523 were screened for EBV-DNA with 796 samples. One hundred twenty-eight (67.4%) of 190 tested patients presented at least 1 positive sample for EBV. Older age, the use of sirolimus, everolimus, and steroids were associated with EBV-DNA positivity in the univariate analysis. Nine (1.7%) of 523 patients had PTLD. Incidence rate of PTLD in the KT cohort was 0.19/100 person year follow-up (95% confidence interval, 0.09-0.37). One of 9 patients developed early PTLD and was a high-risk patient. Only this PTLD case was positive for EBV. No PTLD case had an EBV-VL superior to 4000 copies/mL. Conclusions. Our results suggest that the keystone of PTLD diagnosis is the clinical suspicion. Our study suggests that, in line with guidelines, EBV-VL assays may be avoided in low-risk patients in the absence of a strong clinical PTLD suspicion without increasing patients' risk of developing PTLD. This represents a safe and cost-saving clinical strategy for our center

2017 - Detection of Fusarium-specific T cells in hematologic patients with invasive fusariosis [Articolo su rivista]
Vallerini, Daniela; Forghieri, Fabio; Lagreca, Ivana; Riva, Giovanni; Barozzi, Patrizia; Quadrelli, Chiara; Morselli, Monica; Bresciani, Paola; Cuoghi, Angela; Coluccio, Valeria; Maccaferri, Monica; Paolini, Ambra; Colaci, Elisabetta; Marasca, Roberto; Narni, Franco; Cellini, Monica; Beauvais, Anne; Latgè, Jean Paul; Romani, Luigina; Iughetti, Lorenzo; Comoli, Patrizia; Campioli, Daniele; Trenti, Tommaso; Luppi, Mario; Potenza, Leonardo
abstract

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2017 - Liposomal amphotericin B (AmBisome®) at beginning of its third decade of clinical use [Articolo su rivista]
Aversa, Franco; Busca, Alessandro; Candoni, Anna; Cesaro, Simone; Girmenia, Corrado; Luppi, Mario; Nosari, Anna Maria; Pagano, Livio; Romani, Luigina; Rossi, Giuseppe; Venditti, Adriano; Novelli, Andrea
abstract

Haematologists have been using liposomal amphotericin-B (L-AMB) since 1993 and despite the introduction of several novel antifungal agents over the past decade, their propensity to prescribe L-AMB remains unchanged. Although antifungal guidelines strongly recommend voriconazole as the drug of choice in the treatment of probable and proven invasive fungal disease (IFD), L-AMB is widely used in the real life because of its several advantages in terms of wide anti-mould spectrum, tolerability and efficacy. Furthermore, the concept that L-AMB is endowed with immune-modulating effects, which may have a role in fighting IFD, represent another reason for its use in patients who do not tolerate an excess of inflammation. Finally, given its tolerability and safety, L-AMB is an ideal partner for combining therapy, particularly with echinocandins with which shares the immunological properties that result in a synergistic effect.

2017 - Macitentan, a double antagonist of endothelin receptors, efficiently impairs migration and microenvironmental survival signals in chronic lymphocytic leukemia [Articolo su rivista]
Maffei, Rossana; Fiorcari, Stefania; Vaisitti, Tiziana; Martinelli, Silvia; Benatti, Stefania; Debbia, Giulia; Rossi, Davide; Zucchini, Patrizia; Potenza, Leonardo; Luppi, Mario; Gaidano, Gianluca; Deaglio, Silvia; Marasca, Roberto
abstract

The crosstalk between chronic lymphocytic leukemia (CLL) cells and tumor microenvironment is essential for leukemic clone maintenance, supporting CLL cells survival, proliferation and protection from drug-induced apoptosis. Over the past years, the role of several soluble factors involved in these processes has been studied. CLL cells express higher levels of endothelin 1 (ET-1) and ETA receptor as compared to normal B cells. Upon ET-1 stimulation, CLL cells improve their survival and proliferation and reduce their sensitivity to the phosphoinositide-3-kinase d inhibitor idelalisib and to fludarabine. Here, we demonstrate that CLL cells express not only ETA receptor but also ETB receptor. ET-1 acts as a homing factor supporting CLL cells migration and adhesion to microenvironmental cells. In addition, ET-1 stimulates a pro-angiogenic profile of CLL cells increasing VEGF expression through hypoxia-inducible factor-1 (HIF-1α) accumulation in CLL cells. Macitentan, a specific dual inhibitor of ETAand ETBreceptors, targets CLL cells affecting leukemic cells migration and adhesion and overcoming the pro-survival and proliferation signals mediated by microenvironment. Furthermore, macitentan cooperates with ibrutinib inhibiting the BCR pathway and with ABT-199 disrupting BCL2 pathway. Our data describe the biological effects of a new drug, macitentan, able to counteract essential processes in CLL pathobiology as survival, migration, trafficking and drug resistance. These findings envision the possibility to interfere with ET receptors activity using macitentan as a possible novel therapeutic strategy for CLL patients.

2017 - Monoclonal Gammopathy of Undetermined Significance After Kidney Transplantation: Single-Center Experience [Articolo su rivista]
Alfano, Gaetano; Fontana, Francesco; Colaci, Elisabetta; Messerotti, Andrea; Bettelli, Francesca; Potenza, Leonardo; Luppi, Mario; Cappelli, Gianni
abstract

Background: Monoclonal gammopathy of undetermined significance (MGUS) is an asymptomatic premalignant plasma cell disorder. Prevalence and clinical outcomes of MGUS in kidney transplant (KT) recipients have been previously reported in few studies with conflicting results. Methods: We conducted a retrospective study in a population of 548 KT recipients transplanted between 1998 and 2015. Results: Thirty-nine (8.1%) subjects developed MGUS after KT. At diagnosis of MGUS, the average age was 52 ± 9.2 years, and 23% of the patients were younger than 50 years. Occurrence of MGUS was not influenced by age and sex. After a mean follow-up of 7.8 years, only 1 (2.5%) patient progressed to multiple myeloma. We found no differences in the incidence of solid and hematological malignancies, serious infections, graft failure, and mortality between KT patients with MGUS and a matched cohort of KT recipients without MGUS. The MGUS group had a significantly higher prevalence of monoclonal B cell lymphocytosis, premalignant condition poorly described in KT recipients. Prior history of glomerulonephritis or interstitial nephritis, as cause of renal failure, represented the only predictive factor for MGUS development. Conclusions: MGUS is a premalignant disorder frequently encountered in KT recipients. We found no differences in clinical outcomes between MGUS patients and KT controls.

2017 - More About the Risk of Ibrutinib-associated Bleeding [Articolo su rivista]
Paolini, Ambra; Coluccio, Valeria; Luppi, Mario; Marietta, Marco
abstract

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2017 - Multicenter Prospective Study for Laboratory Diagnosis of HHV8 Infection in Solid Organ Donors and Transplant Recipients and Evaluation of the Clinical Impact After Transplantation [Articolo su rivista]
Chiereghin, Angela; Barozzi, Patrizia; Petrisli, Evangelia; Piccirilli, Giulia; Gabrielli, Liliana; Riva, Giovanni; Potenza, Leonardo; Cappelli, Gianni; de Ruvo, Nicola; Libri, Irene; Maggiore, Umberto; Morelli, Maria Cristina; Potena, Luciano; Todeschini, Paola; Gibertoni, Dino; Labanti, Manuel; Sangiorgi, Gabriela; la Manna, Gaetano; Pinna, Antonio Daniele; Luppi, Mario; Lazzarotto, Tiziana
abstract

BACKGROUND: We performed serological and molecular pretransplant screening in solid organ transplant (SOT) donors and recipients in north-central Italy and a surveillance program for human herpes virus 8 (HHV8) infection after transplant, aiming to establish an optimal management of HHV8 infection in SOT recipients. METHODS: For pretransplant HHV8 screening in both donors and recipients, 6 serological (4 indirect immunofluorescent assays (IFA) and 2 enzyme-linked immunosorbent assays (ELISA) - both HHV8 lytic and latent antigen-based) and 2 molecular assays were used. A reference standard to identify HHV8-positive patients was defined by at least 2 positive assays. All transplant patients at risk to develop HHV8-related disease underwent virological posttransplant monitoring by quantitative real-time PCR assay. RESULTS: HHV8 seroprevalence was 4% (10/249) in donors and 18% (93/517) in organ recipients. The best performance was obtained by 2 lytic antigen-based IFAs that showed almost perfect agreement to the reference standard (0.943 and 0.931 Cohen’s kappa). HHV8-DNA was detected in 6.8% and 2.9% of HHV8-seropositive donor samples by in-house nested PCR and quantitative real-time PCR assays, respectively. After transplant, 3 out of 12 (25%) HHV8-mismatch patients (seropositive donor/seronegative recipient) developed a primary infection, 1 of whom developed a lethal nonmalignant illness. Two out of 93 HHV8-seropositive recipients (2.1%) had viral replication in posttransplant period, 1 of whom developed Kaposi’s sarcoma. CONCLUSIONS: Serological assays, specifically lytic IFAs, were the best methodological approach to identify HHV8-infected SOT donors and recipients. A very low incidence (1.9%) of posttransplant HHV8-related disease was observed.

2017 - Randomized comparison of liposomal amphotericin B versus placebo to prevent invasive mycoses in acute lymphoblastic leukaemia [Articolo su rivista]
Cornely, O. A.; Leguay, T.; Maertens, J.; Vehreschild, M. J. G. T.; Anagnostopoulos, A.; Castagnola, C.; Verga, L.; Rieger, C.; Kondakci, M.; Harter, G.; Duarte, R. F.; Allione, B.; Cordonnier, C.; Heussel, C. P.; Morrissey, C. O.; Agrawal, S. G.; Peter Donnelly, J.; Bresnik, M.; Hawkins, M. J.; Garner, W.; Gokbuget, N.; Jarchum, G.; Dictar, M.; Ramirez Borga, S.; Valledor, A.; Knoebl, P.; Greil, R.; Linkesch, W.; Sill, H.; De Prijck, B.; Sonet, A.; Theunissen, K.; Selleslag, D.; Vargas Schwarzbold, A.; Nucci, M. L. M.; Lopes de Castro Lobo, C.; Fogliatto, L.; Bonmati, C.; Turlure, P.; Herbrecht, R.; Thiebaut, A.; Michallet, M.; Leguay, T.; Egerer, G.; Silling, G.; Pfreundschuh, M.; Hasenkamp, J.; Kraemer, D. M.; Topp, M.; Heinz, W.; Junghanss, C.; Schaich, M. A.; Parmentier, S.; Roellig, C.; Beck, H. J.; Huttmann, A.; Mousset, S.; Duenzinger, U. N.; Schwartz, S.; Haerter, G.; Kondakci, M.; Ostermann, H.; Rieger, C.; Cornely, O. A.; Vehreschild, M. J. G. T.; Tsirigotis, P.; Matsouka, P.; Angelopoulou, M. K.; Karakantza, M.; Spyridonidis, A.; Anagnostopoulos, A.; Kolomansky, A.; Moses, A.; Horowitz, N.; Rahav, G.; Aversa, F.; Velardi, A.; Pagano, L.; Gentile, G.; Gobbi, M.; Luppi, M.; Nosari, A. M.; Rambaldi, A.; Candoni, A.; Marbello, L.; Rossi, G.; Pogliani, E.; Allione, B.; Castagnola, C.; Moreira, I.; Nunes, A.; Botelho de Sousa, A.; Rubio Tejero, A. I.; Vallejo, C.; Vazquez, L.; Besalduch Vidal, J.; Gomez-Garcia de Soria, V.; Jurado Chacon, M.; Gonzalez Campos, J.; Olavarria, E.; Barba, P.; de la Serna Torroba, J.; Duarte, R.; Heim, D.; Zimmerli, S.; Gerber, B.; Akova, M.; Bolaman, A. Z.; Tabak, F.; Akan, H.; Senol, E.
abstract

Objectives: To prevent invasive fungal disease (IFD) in adult patients undergoing remission-induction chemotherapy for newly diagnosed acute lymphoblastic leukaemia (ALL). Patients and methods: In a double-blind multicentre Phase 3 study, patients received prophylactic liposomal amphotericin B (L-AMB) at 5 mg/kg intravenously or placebo twice weekly in a 2:1 random allocation during remission-induction treatment. The primary endpoint was the development of proven or probable IFD. Secondary endpoints included those focused on the safety and tolerability of prophylactic L-AMB. Results: Three hundred and fifty-five patients from 86 centres in Europe and South America received at least one dose of L-AMB (n = 237) or placebo (n = 118). Rates of proven and probable IFD assessed independently were 7.9%(18/228) in the L-AMB group and 11.7%(13/111) in the placebo group (P = 0.24). Rates of possible IFD were 4.8% (11/228) in the L-AMB and 5.4% (6/111) in the placebo group (P = 0.82). The remission-induction phase was a median of 22 days for both groups. Overall mortality was similar between the groups: 7.2% (17/237) for L-AMB and 6.8% (8/118) for placebo (P = 1.00). Hypokalaemia and creatinine increase were significantly more frequent with L-AMB. Conclusions: The IFD rate among adult patients undergoing remission-induction chemotherapy for newly diagnosed ALL was 11.7%in the placebo group, andwas not significantly different in patients receiving L-AMB, suggesting that the L-AMB regimen studied is not effective as prophylaxis against IFD. The IFD rate appears higher than previously reported, warranting further investigation. Tolerability of L-AMB was what might be expected. Further studies are needed to determine the optimal antifungal strategy during remission-induction chemotherapy of ALL.

2017 - Successful Treatment of KSHV Inflammatory Cytokine Syndrome (KICS) after Kidney-Liver Transplant: Correlations with HHV8 miRNome and Specific T-Cell Response [Articolo su rivista]
Mularoni, Alessandra; Gallo, Alessia; Riva, Giovanni; Barozzi, Patrizia; Miele, Monica; Cardinale, Giovanni; Vizzini, Giovanni; Volpes, Riccardo; Grossi, Paolo; Di Carlo, Daniele; Luca, Angelo; Trenti, Tommaso; Luppi, Mario; Conaldi, Pier Giulio
abstract

In post-transplant patients, HHV-8/KSHV infection is known to cause aggressive tumors, as well as severe non-neoplastic complications. These latter syndromes are driven by HHV-8/KSHV lytic reactivations and related hyper-inflammatory host responses, typically characterized by high viral loads, elevated levels of cytokines and other inflammation biomarkers, cytopenia, organ failure, high fever, and worsening conditions (with no evidence of B-cell neoplasias). These disorders are associated with a high mortality rate, often due to lack of prompt diagnosis, effective therapeutic approaches, and adequate follow up. Indeed, these features resemble most of those defining the so-called KSHV inflammatory cytokine syndrome (KICS), which was recently recognized in HIV-positive patients. Here we describe, for the first time, a case of KICS-like non-neoplastic recurrent complication occurring in an HIV-negative post-transplant patient, which was successfully treated by combination of anti-CD20 monoclonal therapy, antivirals, and modification of immunosuppressive regimen. In addition to clinical and laboratory findings collected during 3-year-long follow up, we also report novel experimental data on HHV-8-specific T-cell dynamics and circulating miRNA profile, showing correlations with clinical course and other laboratory markers (including viral load, CRP and cytokine levels), thus providing useful information about abnormal cellular and cytokine dynamics underlying of HHV-8-associated inflammatory disorders in post-transplant patients. This article is protected by copyright. All rights reserved.

2017 - The expression of endothelin-1 in chronic lymphocytic leukemia is controlled by epigenetic mechanisms and extracellular stimuli. [Articolo su rivista]
Martinelli, Silvia; Maffei, Rossana; Fiorcari, Stefania; Quadrelli, Chiara; Zucchini, Patrizia; Benatti, Stefania; Potenza, Leonardo; Luppi, Mario; Marasca, Roberto
abstract

Endothelin-1 (ET-1) is a hormone peptide widely expressed and is involved in several biological processes, important not only for normal cell function but also for tumor development, including cell proliferation, invasion, metastasis, angiogenesis and osteogenesis. In accordance, ET-1 was already shown to contribute to the growth and progression of many different solid cancers. We recently demonstrated that ET-1 has a role in the pathogenesis of chronic lymphocytic leukemia (CLL) where it is abnormally expressed. In the context of this malignancy, ET-1 is able to mediate survival, drug-resistance and growth signals in leukemic cells. Previous studies, not conducted in CLL, have shown that ET-1 regulatory mechanisms are numerous and cell specific. Here, we valued the expression of ET-1 in CLL, in relation to DNA methylation but also in response to stimulation of some important pathways for the dialogue between CLL and microenvironment. We found that a high methylation of ET-1 first intron affects the basal expression of ET-1 in CLL. Moreover, we showed that the activation of CD40 or Toll-like receptor (TLR) by extracellular stimuli produces an augment of ET-1 level in CLL cells. Finally, we demonstrated the fundamental role of NF-kB signalling pathway in promoting and maintaining ET-1 expression in CLL cells, both in basal conditions and after CD40 activation.

2017 - The importance of cytogenetic and molecular analyses in eosinophilia-associated myeloproliferative neoplasms: an unusual case with normal karyotype and TNIP1- PDGFRB rearrangement and overview of PDGFRB partner genes [Articolo su rivista]
Maccaferri, Monica; Pierini, V.; Di Giacomo, D.; Zucchini, Patrizia; Forghieri, Fabio; Bonacorsi, G.; Paolini, Ambra; Quadrelli, Chiara; Giacobbi, F.; Fontana, Francesco; Cappelli, Gianni; Potenza, Leonardo; Marasca, Roberto; Luppi, Mario; Mecucci, C.
abstract

Fusion genes derived from the platelet-derived growth factor receptor beta (PDGFRB) or alpha (PDGFRA) play an important role in the pathogenesis of eosinophilia-associated myeloproliferative neoplasms (Eo-MPN). Here, we would like to report a case of Eo-MPN with normal karyotype and rearrangement of TNIP1-PDGFRB, with unusual clinical presentation, which delayed the diagnosis and initiation of an appropriate therapy. We would also like to provide a review of the so far reported PDGFRB fusion partners in myeloid neoplasms, either myeloproliferative or myelodysplastic, summarizing clinical features and response to imatinib

2016 - A sequential approach with imatinib, chemotherapy and transplant for adult Ph+ acute lymphoblastic leukemia. Final results of the GIMEMA LAL 0904 study [Articolo su rivista]
Chiaretti, Sabina; Vitale, Antonella; Vignetti, Marco; Piciocchi, Alfonso; Fazi, Paola; Elia, Loredana; Falini, Brunangelo; Ronco, Francesca; Ferrara, Felicetto; De Fabritiis, Paolo; Luppi, Mario; La Nasa, Giorgio; Tedeschi, Alessandra; Califano, Catello; Fanin, Renato; Dore, Fausto; Mandelli, Franco; Meloni, Giovanna; Foa', Robin
abstract

In the GIMEMA LAL 0904 protocol, adult Ph+ acute lymphoblastic leukemia patients were treated with chemotherapy for induction and consolidation, followed by maintenance with imatinib. The protocol was subsequently amended and imatinib was incorporated in the induction and post-remission phase together with chemotherapy. Due to the toxicity of this combined approach, the protocol was further amended to a sequential scheme based on imatinib plus steroids as induction, followed by consolidation with chemotherapy plus imatinib and, when applicable, by a hematopoietic stem cell transplant. Fifty-one patients (median age: 45.9 years) were enrolled in the final sequential protocol, hereby reported. At the end of induction (day +50), 96% of evaluable patients (n=49) achieved a complete hematologic remission; after consolidation, all were in complete hematologic remission. No deaths in induction were recorded. Overall survival and disease-free survival at 60 months are 48.8% and 45.8%, respectively. At day +50 (end of the imatinib induction), a log-reduction of BCR-ABL1 levels &gt;1.3 was associated with a significantly more prolonged disease-free survival (55.6%, C.I. 95%: 39.0-79.3 vs 20%, C.I. 95%: 5.8-69.1; p=0.03), overall survival (59.1%, C.I. 95%: 42.3-82.6 vs 20%, C.I. 95%: 5.8-69.1, p=0.02) and lower relapse incidence (20.5%, C.I. 95%: 7.2-38.6 vs 60.0%, C.I. 95%: 21.6-84.3, p=0.01). Mean BCR-ABL1 levels remained significantly higher in patients who subsequently relapsed. Finally, BCR-ABL1p190 patients showed a significantly faster molecular response than BCR-ABL1p210 patients (p=0.023). Thought the study was not powered to evaluate the role of allogeneic stem cell transplant, allografting positively impacted on overall survival and disease-free survival. A sequential approach with imatinib alone in induction, consolidated by chemotherapy plus imatinib followed by a stem cell transplant is a feasible, well-tolerated and effective strategy for adult Ph+ acute lymphoblastic leukemia, leading to the best long-term survival rates so far reported. Trial ID: NCT00458848.

2016 - All-trans retinoic acid (ATRA) in non-promyelocytic acute myeloid leukemia (AML): results of combination of ATRA with low-dose Ara-C in three elderly patients with NPM1-mutated AML unfit for intensive chemotherapy and review of the literature [Articolo su rivista]
Forghieri, Fabio; Bigliardi, Sara; Quadrelli, Chiara; Morselli, Monica; Potenza, Leonardo; Paolini, Ambra; Colaci, Elisabetta; Barozzi, Patrizia; Zucchini, Patrizia; Riva, Giovanni; Vallerini, Daniela; Lagreca, Ivana; Marasca, Roberto; Narni, Franco; Venditti, Adriano; Martelli, Maria Paola; Falini, Brunangelo; Lo Coco, Francesco; Amadori, Sergio; Luppi, Mario
abstract

Based upon the clinical behavior of three patients, we suggest that the combination of low-dose Ara-C and all-trans retinoic acid may potentially be effective in some elderly patients, unfit for intensive chemotherapy, affected with NPM1-mutated acute myeloid leukemia without FLT3 mutations, warranting perspective clinical studies in these selected patients.

2016 - CRLF2 overexpression identifies an unfavourable subgroup of adult B-cell precursor acute lymphoblastic leukemia lacking recurrent genetic abnormalities [Articolo su rivista]
Chiaretti, Sabina; Brugnoletti, Fulvia; Messina, Monica; Paoloni, Francesca; Fedullo, Anna Lucia; Piciocchi, Alfonso; Elia, Loredana; Vitale, Antonella; Mauro, Elisa; Ferrara, Felicetto; De Fabritiis, Paolo; Luppi, Mario; Ronco, Francesca; De Propris, Maria Stefania; Raponi, Sara; Kronnie, Geertruy te; Vignetti, Marco; Guarini, Anna; Foà, Robin
abstract

Background: A deregulated CRLF2 (d-CRLF2) expression was described in B-cell acute lymphoblastic leukemia without recurrent fusion genes (B-NEG ALL). While the role of d-CRLF2 in children has been extensively described, little is known about its role and impact in adult ALL. Methods: Expression levels of CRLF2 were evaluated by quantitative real-time PCR in 102 newly-diagnosed adult B-NEG ALL and correlated with the clinico-biological characteristics and outcome. Incidence and clinical impact of the P2RY8/CRLF2 transcript was also assessed. Results: High CRLF2 levels, as continuous variable, were significantly associated with hyperleucocytosis (p = 0.0002) and thrombocytopenia (p = 0.005); when a cut-point at δCt ≤ 8 was applied, 35 cases (34.3%), mostly males (80%), proved positive for CRLF2 expression. High CRLF2 levels, as continuous or categorical variable, were associated with a worse disease-free (p = 0.003 and p = 0.015) and overall survival (p = 0.017 and 0.0038). Furthermore, when CRLF2 was analyzed as a categorical variable, a high statistical association was found with IKZF1 deletion and mutations in the JAK/STAT pathway (p = 0.001 and p &lt; 0.0001, respectively). Finally, the P2RY8/CRLF2 transcript, identified in 8/102 patients (7.8%), was associated with a poor outcome. Conclusions: In adult B-NEG ALL, high CRLF2 expression is associated with distinct clinico-biological features and an unfavourable prognosis in both univariate and multivariate analysis; similarly, P2RY8/CRLF2 positivity correlates with a poor outcome. The quantification of CRLF2 is an important prognostic marker in adult B-lineage ALL without known genetic lesions.

2016 - Chronic and recurrent benign lymphadenopathy without constitutional symptoms associated with human herpesvirus-6B reactivation [Articolo su rivista]
Forghieri, Fabio; Luppi, Mario; Barozzi, Patrizia; Riva, Giovanni; Monica, Morselli; Bigliardi, Sara; Quadrelli, Chiara; Vallerini, Daniela; Maccaferri, Monica; Coluccio, Valeria; Paolini, Ambra; Colaci, Elisabetta; Goretta, Bonacorsi; Maiorana, Antonino; Sara, Tagliazucchi; Fabio, Rumpianesi; Mattioli, Francesco; Presutti, Livio; Gelmini, Roberta; Cermelli, Claudio; Giulio, Rossi; Patrizia, Comoli; Marasca, Roberto; Narni, Franco; Potenza, Leonardo
abstract

Chronic/recurrent behaviour may be encountered in some distinct atypical or malignant lymphoproliferations, while recurrences are not generally observed in reactive/benign lymphadenopathies. We retrospectively anal- ysed a consecutive series of 486 human immunodeficiency virus-negative adults, who underwent lymphadenectomy. Neoplastic and benign/reactive histopathological pictures were documented in 299 (61 5%) and 187 (38 5%) cases, respectively. Of note, seven of the 111 (6 3%) patients with benign lymphadenopathy without well-defined aetiology, showed chronic/ recurrent behaviour, without constitutional symptoms. Enlarged lymph nodes were round in shape and hypoechoic, mimicking lymphoma. Reac- tive follicular hyperplasia and paracortical expansion were observed. Human herpesvirus (HHV)-6B positive staining in follicular dendritic cells (FDCs) was documented in all seven patients. Serological, molecular and immunological examinations suggested HHV-6B reactivation. Among the remaining 104 cases with reactive lymphoid hyperplasia in the absence of well-known aetiology and without recurrences, positivity for HHV-6B on FDCs was found in three cases, whereas in seven further patients, a scanty positivity was documented in rare, scattered cells in inter-follicular regions. Immunohistochemistry for HHV-6A and HHV-6B was invariably negative on 134 lymph nodes, with either benign pictures with known aetiology or malignant lymphoproliferative disorders, tested as further controls. Future studies are warranted to investigate a potential association between HHV- 6B reactivation and chronic/recurrent benign lymphadenopathy.

2016 - Circulating functional T cells specific to human herpes virus 6 (HHV6) antigens in individuals with chromosomally integrated HHV6 [Articolo su rivista]
Barozzi, Patrizia; Riva, Giovanni; Vallerini, Daniela; Quadrelli, Chiara; Lagreca, Ivana; Eccheli, Roberta; Forghieri, Fabio; Coluccio, Valeria; Maccaferri, Monica; Paolini, Ambra; Colaci, Elisabetta; Morselli, Monica; Marasca, Roberto; Narni, Franco; Potenza, Leonardo; Luppi, Mario; Comoli, Patrizia; Campioli, Daniele; Trenti, Tommaso
abstract

Circulating functional T cells specific to human herpes virus 6 (HHV6) antigens in individuals with chromosomally integrated HHV6

2016 - Common Genetic Polymorphisms within NFκB-Related Genes and the Risk of Developing Invasive Aspergillosis [Articolo su rivista]
Lupiañez, Carmen B; Villaescusa, María T; Carvalho, Agostinho; Springer, Jan; Lackner, Michaela; Sánchez Maldonado, José M; Canet, Luz M; Cunha, Cristina; Segura Catena, Juana; Alcazar Fuoli, Laura; Solano, Carlos; Fianchi, Luana; Pagano, Livio; Potenza, Leonardo; Aguado, José M; Luppi, Mario; Cuenca Estrella, Manuel; Lass Flörl, Cornelia; Einsele, Hermann; Vázquez, Lourdes; Ríos Tamayo, Rafael; Loeffler, Jurgen; Jurado, Manuel; Sainz, Juan
abstract

Invasive Aspergillosis (IA) is an opportunistic infection caused by Aspergillus, a ubiquitously present airborne pathogenic mold. A growing number of studies suggest a major host genetic component in disease susceptibility. Here, we evaluated whether 14 single-nucleotide polymorphisms within NFκB1, NFκB2, RelA, RelB, Rel, and IRF4 genes influence the risk of IA in a population of 834 high-risk patients (157 IA and 677 non-IA) recruited through a collaborative effort involving the aspBIOmics consortium and four European clinical institutions. No significant overall associations between selected SNPs and the risk of IA were found in this large cohort. Although a hematopoietic stem cell transplantation (HSCT)-stratified analysis revealed that carriers of the IRF4 rs12203592T/T genotype had a six-fold increased risk of developing the infection when compared with those carrying the C allele (ORREC = 6.24, 95%CI 1.25-31.2, P = 0.026), the association of this variant with IA risk did not reach significance at experiment-wide significant threshold. In addition, we found an association of the IRF4AATC and IRF4GGTC haplotypes (not including the IRF4 rs12203592T risk allele) with a decreased risk of IA but the magnitude of the association was similar to the one observed in the single-SNP analysis, which indicated that the haplotypic effect on IA risk was likely due to the IRF4 rs12203592 SNP. Finally, no evidence of significant interactions among the genetic markers tested and the risk of IA was found. These results suggest that the SNPs on the studied genes do not have a clinically relevant impact on the risk of developing IA.

2016 - Evaluation of serum (1 → 3)-β-D-glucan clinical performance: kinetic assessment, comparison with galactomannan and evaluation of confounding factors [Articolo su rivista]
Pini, Pietro; Bettua, C; Orsi, Carlotta Francesca; Venturelli, C; Forghieri, F; Bigliardi, S; Faglioni, L; Luppi, Fabrizio; Serio, L; Codeluppi, M; Luppi, Mario; Mussini, Cristina; Girardis, Massimo; Blasi, Elisabetta
abstract

Purpose We investigated the clinical performance of (1 → 3)-β-d-glucan (BG), as an early marker of invasive fungal infections (IFI), in different clinical settings. Methods BG serum levels were assessed by Fungitell (Associates of Cape Cod, Inc), in parallel with galactomannan (GM) when requested by clinicians. By a prospective monocentric study, 270 episodes at risk or with suspect of IFI were enrolled, namely 58 proven-probable invasive aspergillosis (IA), 27 proven invasive candidiasis (IC), 11 possible IC, 16 P.jirovecii pneumonia (PJP), 4 episodes of other IFI and 154 non-IFI controls. Results We found that (a) the BG overall sensitivity, specificity, positive predictive value and negative predictive value (NPV) were 87.9, 80.5, 76.7 and 89.9 %, respectively; (b) the highest sensitivity was found in the IC groups, followed by PJP, IA and other IFI groups; (c) an association was observed between BG kinetics and patients outcome; (d) in the IA episodes, the combination of BG or GM vs GM alone increased sensitivity from 60.0 to 83.3 % in the haematological patients; (e) false-positive BG results were related to Gram-negative infections or infusion of polyclonal IgM-enriched immunoglobulins, where high levels of BG were indeed detected. Conclusion Besides strengthening its overall good clinical performance, we provide evidence that serum BG correlates with clinical outcome and that, once used in combination with GM, BG allows to enhance IFI diagnosis rate. The high sensitivity and NPV, observed in the Intensive Care Unit setting, open to BG validation as a marker for assessment of antifungal treatment.

2016 - Ibrutinib modifies the function of monocyte/macrophage population in chronic lymphocytic leukemia [Articolo su rivista]
Fiorcari, Stefania; Maffei, Rossana; Audrito, Valentina; Martinelli, Silvia; Hacken, Elisa Ten; Zucchini, Patrizia; Grisendi, Giulia; Potenza, Leonardo; Luppi, Mario; Burger, Jan A; Deaglio, Silvia; Marasca, Roberto
abstract

In lymphoid organs, nurse-like cells (NLCs) show properties of tumor-associated macrophages, playing a crucial role in chronic lymphocytic leukemia (CLL) cell survival. Ibrutinib, a potent inhibitor of Bruton's tyrosine kinase (BTK), is able to counteract pro-survival signals in CLL cells. Since the effects on CLL cells have been studied in the last years, less is known about the influence of ibrutinib on NLCs properties. We sought to determine how ibrutinib modifies NLCs functions focusing on the balance between immunosuppressive and inflammatory features. Our data show that ibrutinib targets BTK expressed by NLCs modifying their phenotype and function. Treatment with ibrutinib reduces the phagocytic ability and increases the immunosuppressive profile of NLCs exacerbating the expression of M2 markers. Accordingly, ibrutinib hampers LPS-mediated signaling, decreasing STAT1 phosphorylation, while allows IL-4-mediated STAT6 phosphorylation. In addition, NLCs treated with ibrutinib are able to protect CLL cells from drug-induced apoptosis partially through the secretion of IL-10. Results from patient samples obtained prior and after 1 month of treatment with ibrutinib show an accentuation of CD206, CD11b and Tie2 in the monocytic population in the peripheral blood. Our study provides new insights into the immunomodulatory action of ibrutinib on monocyte/macrophage population in CLL.

2016 - Lenalidomide in chronic lymphocytic leukemia: The present and future in the era of tyrosine kinase inhibitors [Articolo su rivista]
Maffei, Rossana; Colaci, Elisabetta; Fiorcari, Stefania; Martinelli, Silvia; Potenza, Leonardo; Luppi, Mario; Marasca, Roberto
abstract

Lenalidomide is an immunomodulatory agent (IMiD) clinically active in chronic lymphocytic leukemia (CLL), both in heavily pre-treated patients and upfront. Lenalidomide has a unique mechanism of action in CLL. Its efficacy relies on a multifactorial mode-of-action (MOA), comprising a plethora of immunomodulatory actions, the disruption of mutualistic interactions inside CLL microenvironment and direct effects against leukemic cells. In the last few years, a number of new and highly effective drugs appeared in the scenario of CLL therapeutic options, i.e. tyrosine kinase inhibitors (TKIs), showing a good safety profile and impressive clinical response, also in high-risk patients. In this review, we describe the data from clinical studies about lenalidomide efficacy in CLL and we critically dissect the different mechanisms of action of this drug. We point the attention on open issues, including drug dosage and administration schedule, prediction of clinical response to lenalidomide, and combination therapeutic strategies. This overview would be useful to envision a possible role of lenalidomide in the treatment flow-chart of CLL, exploiting its peculiar MOA and also exploring the possible synergetic effect with new drugs.

2016 - Mucorales-specific T cells in patients with hematologic malignancies [Articolo su rivista]
Potenza, Leonardo; Vallerini, Daniela; Barozzi, Patrizia; Riva, Giovanni; Gilioli, Andrea; Forghieri, Fabio; Candoni, Anna; Cesaro, Simone; Quadrelli, Chiara; Maertens, Johan; Rossi, Giulio; Morselli, Monica; Codeluppi, Mauro; Mussini, Cristina; Colaci, Elisabetta; Messerotti, Andrea; Paolini, Ambra; Maccaferri, Monica; Fantuzzi, Valeria; DEL GIOVANE, Cinzia; Stefani, Alessandro; Morandi, Uliano; Maffei, Rossana; Marasca, Roberto; Narni, Franco; Fanin, Renato; Comoli, Patrizia; Romani, Luigina; Beauvais, Anne; Viale, Pier Luigi; Latgè, Jean Paul; Lewis, Russell E.; Luppi, Mario
abstract

Background: Invasive mucormycosis (IM) is an emerging life-threatening fungal infection. It is difficult to obtain a definite diagnosis and to initiate timely intervention. Mucorales-specific T cells occur during the course of IM and are involved in the clearance of the infection. We have evaluated the feasibility of detecting Mucorales-specific T cells in hematological patients at risk for IM, and have correlated the detection of such cells with the clinical conditions of the patients. Methods and Findings: By using an enzyme linked immunospot assay, the presence of Mucorales-specific T cells in peripheral blood (PB) samples has been investigated at three time points during highdose chemotherapy for hematologic malignancies. Mucorales-specific T cells producing interferon-γ, interleukin-10 and interleukin-4 were analysed in order to detect a correlation between the immune response and the clinical picture. Twenty-one (10.3%) of 204 patients, accounting for 32 (5.3%) of 598 PB samples, tested positive for Mucorales-specific T cells. Two groups could be identified. Group 1, including 15 patients without signs or symptoms of invasive fungal diseases (IFD), showed a predominance of Mucorales-specific T cells producing interferon-gamma. Group 2 included 6 patients with a clinical picture consistent with invasive fungal disease (IFD):2 cases of proven IM and 4 cases of possible IFD. The proven patients had significantly higher number of Mucorales-specific T cells producing interleukin-10 and interleukin-4 and higher rates of positive samples by using derived diagnostic cut-offs when compared with the 15 patients without IFD. Conclusions: Mucorales-specific T cells can be detected and monitored in patients with hematologic malignancies at risk for IM. Mucorales-specific T cells polarized to the production of T helper type 2 cytokines are associated with proven IM and may be evaluated as a surrogate diagnostic marker for IM.

2016 - PEGylated siRNA lipoplexes for silencing of BLIMP-1 in Primary Effusion Lymphoma: In vitro evidences of antitumoral activity [Articolo su rivista]
Belletti, Daniela; Tosi, Giovanni; Forni, Flavio; Lagreca, Ivana; Barozzi, Patrizia; Pederzoli, Francesca; Vandelli, Maria Angela; Riva, Giovanni; Luppi, Mario; Ruozi, Barbara
abstract

Silencing of the B lymphocyte-induced maturation protein 1 (Blimp-1), a pivotal transcriptional regulator during terminal differentiation of B cells into plasma cells with siRNAs is under investigation as novel therapeutic approach in Primary Effusion Lymphoma (PEL), a HHV-8 related and aggressive B cell Lymphoma currently lacking of an efficacious therapeutic approach. The clinical application of small interfering RNA (siRNA) in cancer therapy is limited by the lack of an efficient systemic siRNA delivery system. In this study we aim to develop pegylated siRNA lipoplexes formed using the cationic lipid DOTAP and DSPE-PEG2000, capable to effectively stabilize anti-Blimp-1 siRNA and suitable for systemic administration. Two types of pegylated lipoplexes using a classic (C-PEG Lipoplexes) or a post-pegylation method (P-PEG-Lipoplexes) were formulated and compared in their physicochemical properties (size, zeta potential, morphology and structure) and efficiency on PEL cell lines. A stable siRNAs protection was obtained with post pegylation approach (2% molar of DSPE-PEG2000 with respect to lipid) resulting in structures with diameters of 300 nm and a complexation efficiency higher that 80% (0.08 nmol/10 nmol of lipid). In vitro studies on PEL cell lines suggested that empty liposomes were characterized by a low cell toxicity also after PEG modification (cell viability and cell density over 85% after treatment with 10 μM of lipid). We demonstrated that P-PEG-Lipoplexes were able to significantly reduce the levels of BLIMP-1 protein leading to reduction of viability (less that 15% after transfection with 100 nM of complexed siRNAs) and activation of apoptosis. In vitro efficiency encourages us to further test the in vivo potential of P-PEG-Lipoplexes in PEL therapy.

2016 - Personal history and quality of life in chronic myeloid leukemia patients: a cross-sectional study using narrative medicine and quantitative analysis [Articolo su rivista]
Breccia, Massimo; Graffigna, Guendalina; Galimberti, Sara; Iurlo, Alessandra; Pungolino, Ester; Pizzuti, Michele; Maggi, Alessandro; Falzetti, Franca; Capalbo, Silvana Franca; Intermesoli, Tamara; Maffioli, Margherita; Elena, Chiara; Melosi, Alessandro; Simonetti, Federico; Capochiani, Enrico; Seta, Roberta Della; Pacilli, Matteo; Luppi, Mario; Di Renzo, Nicola; Mastrullo, Lucia; Trabacchi, Elena; Vallisa, Daniele; Rapezzi, Davide; Orlandi, Ester Maria; Gambacorti Passerini, Carlo; Efficace, Fabio; Alimena, Giuliana
abstract

Tyrosine kinase inhibitors (TKIs) drastically changed the outcome of patients diagnosed with chronic myeloid leukemia (CML). Several reports indicated the advantage of continue long-term adherence associated with positive outcome. Therefore, it is important to better understand from the patient's standpoint the experience of living with the disease and the related treatment. OBJECTIVES: In this study, quantitative analysis and narrative medicine were combined to get insights on this issue in a population of 257 patients with CML in chronic phase treated with TKIs (43 % men, with a median age of 58 years, 27 % aged 31-50 years), followed for a median time of 5 years. Sixty-one percent of patients enrolled were treated in first line, whereas 37 % were treated in second line. RESULTS: The results showed more positive perceptions and acceptance in males compared to females, without impact of disease on relationships. Level of positive acceptance was more evident in elderly compared to younger patients, with a close connection with median time from diagnosis. Overall, female patients reported negative perceptions and an impact of disease on family daily living. The majority of patients understood the importance of continue adherence to treatment, with 27 % resulting less adherent (60 % for forgetfulness), even if well informed and supported by his/her physician. DISCUSSION AND CONCLUSIONS: Narrative medicine, in association to quantitative analysis, can help physicians to understand needs of their patients in order to improve communication.

2016 - Polymorphisms in host immunity-modulating genes and risk of invasive aspergillosis: Results from the AspBIOmics Consortium [Articolo su rivista]
Lupiañez, C. B.; Canet, L. M.; Carvalho, A.; Alcazar Fuoli, L.; Springer, J.; Lackner, M.; Segura Catena, J.; Comino, A.; Olmedo, C.; Ríos, R.; Fernández Montoya, A.; Cuenca Estrella, M.; Solano, C.; López Nevot, M. Á.; Cunha, C.; Oliveira Coelho, A.; Villaescusa, T.; Fianchi, L.; Aguado, J. M.; Pagano, L.; López Fernández, E.; Potenza, Leonardo; Luppi, Mario; Lass Flörl, C.; Loeffler, J.; Einsele, H.; Vazquez, L.; Jurado, M.; Sainz, J.
abstract

Recent studies suggest that immune-modulating single-nucleotide polymorphisms (SNPs) influence the risk of developing cancer-related infections. Here, we evaluated whether 36 SNPs within 14 immune-related genes are associated with the risk of invasive aspergillosis (IA) and whether genotyping of these variants might improve disease risk prediction. We conducted a case-control association study of 781 immunocompromised patients, 149 of whom were diagnosed with IA. Association analysis showed that the IL4Rrs2107356 and IL8rs2227307 SNPs (using dbSNP numbering) were associated with an increased risk of IA (IL4Rrs2107356 odds ratio [OR], 1.92; 95% confidence interval [CI], 1.20 to 3.09; IL8rs2227307 OR, 1.73; 95% CI, 1.06 to 2.81), whereas the IL12Brs3212227 and IFNγrs2069705 variants were significantly associated with a decreased risk of developing the infection (IL12Brs3212227 OR, 0.60; 95% CI, 0.38 to 0.96; IFNγrs2069705 OR, 0.63; 95% CI, 0.41 to 0.97). An allogeneic hematopoietic stem cell transplantation (allo-HSCT)-stratified analysis revealed that the effect observed for the IL4Rrs2107356 and IFNγrs2069705 SNPs was stronger in allo-HSCT (IL4Rrs2107356 OR, 5.63; 95% CI, 1.20 to 3.09; IFNγrs2069705 OR, 0.24; 95% CI, 0.10 to 0.59) than in non-HSCT patients, suggesting that the presence of these SNPs renders patients more vulnerable to infection, especially under severe and prolonged immunosuppressive conditions. Importantly, in vitro studies revealed that carriers of the IFNγrs2069705C allele showed a significantly increased macrophage-mediated neutralization of fungal conidia (P = 0.0003) and, under stimulation conditions, produced higher levels of gamma interferon (IFNγ) mRNA (P = 0.049) and IFNγ and tumor necrosis factor alpha (TNF-α) cytokines (P value for 96 h of treatment with lipopolysaccharide [PLPS-96 h], 0.057; P value for 96 h of treatment with phytohemagglutinin [PPHA-96 h], 0.036; PLPS+PHA-96 h = 0.030; PPHA-72 h = 0.045; PLPS+PHA-72 h = 0.018; PLPS-96 h = 0.058; PLPS+PHA-96 h = 0.0058). Finally, we also observed that the addition of SNPs significantly associated with IA to a model including clinical variables led to a substantial improvement in the discriminatory ability to predict disease (area under the concentration-time curve [AUC] of 0.659 versus AUC of 0.564; P-2 log likehood ratio test = 5.2 · 10-4 and P50.000 permutation test = 9.34 · 10-5). These findings suggest that the IFNγrs2069705 SNP influences the risk of IA and that predictive models built with IFNγ, IL8, IL12p70, and VEGFA variants can used to predict disease risk and to implement risk-adapted prophylaxis or diagnostic strategies.

2016 - Randomized trial of low-dose morphine versus weak opioids in moderate cancer pain [Articolo su rivista]
Bandieri, Elena; Romero, Marilena; Ripamonti, Carla Ida; Artioli, Fabrizio; Sichetti, Daniela; Fanizza, Caterina; Santini, Daniele; Cavanna, Luigi; Melotti, Barbara; Conte, Pier Franco; Roila, Fausto; Cascinu, Stefano; Bruera, Eduardo; Tognoni, Gianni; Luppi, Mario
abstract

Purpose: The WHO guidelines on cancer pain management recommend a sequential three-step analgesic ladder. However, conclusive data are lacking as to whether moderate pain should be treated with either step II weak opioids or low-dose step III strong opioids. Patients and Methods: In a multicenter, 28-day, open-label randomized controlled study, adults with moderate cancer pain were assigned to receive either a weak opioid or low-dose morphine. The primary outcome was the number of responder patients, defined as patientswith a 20% reduction in pain intensity on the numerical rating scale. Results: A total of 240 patients with cancer (118 in the low-dose morphine and 122 in the weak-opioid group) were included in the study. The primary outcome occurred in 88.2% of the low-dose morphine and in 57.7% of the weak-opioid group (odds risk, 6.18; 95% CI, 3.12 to 12.24; P&lt;001). The percentage of responder patients was higher in the low-dose morphine group, as early as at 1 week. Clinically meaningful (&gt; 30%) and highly meaningful (&gt; 50%) pain reduction from baseline was significantly higher in the low-dose morphine group (P &lt;.001). A change in the assigned treatment occurred more frequently in the weak-opioid group, because of inadequate analgesia. The general condition of patients, which was based on the Edmonton Symptom Assessment System overall symptom score, was better in the morphine group. Adverse effects were similar in both groups. Conclusion: In patients with cancer and moderate pain, low-dose morphine reduced pain intensity significantly compared with weak opioids, with a similarly good tolerability and an earlier effect.

2016 - Reply to O. Corli et al and M. Lucchesi et al [Articolo su rivista]
Bandieri, Elena; Romero, Marilena; Ripamonti, Carla Ida; Artioli, Fabrizio; Fanizza, Caterina; Cascinu, Stefano; Bruera, Eduardo; Luppi, Mario
abstract

As with the letter from Corli and Ribeiro, the Data Supplement from our article provides readers further points for discussion on the relevant issue of clinically meaningful pain reduction.

2016 - Reversal of the glycolytic phenotype of primary effusion lymphoma cells by combined targeting of cellular metabolism and PI3K/Akt/ mTOR signaling [Articolo su rivista]
Mediani, Laura; Gibellini, Federica; Bertacchini, Jessika; Frasson, Chiara; Bosco, Raffaella; Accordi, Benedetta; Basso, Giuseppe; Bonora, Massimo; Calabrò, Maria Luisa; Mattiolo, Adriana; Sgarbi, Gianluca; Baracca, Alessandra; Pinton, Paolo; Riva, Giovanni; Rampazzo, Enrico; Petrizza, Luca; Prodi, Luca; Milani, Daniela; Luppi, Mario; Potenza, Leonardo; De Pol, Anto; Cocco, Lucio; Capitani, Silvano; Marmiroli, Sandra
abstract

PEL is a B-cell non-Hodgkin lymphoma, occurring predominantly as a lymphomatous effusion in body cavities, characterized by aggressive clinical course, with no standard therapy. Based on previous reports that PEL cells display a Warburg phenotype, we hypothesized that the highly hypoxic environment in which they grow in vivo makes them more reliant on glycolysis, and more vulnerable to drugs targeting this pathway. We established here that indeed PEL cells in hypoxia are more sensitive to glycolysis inhibition. Furthermore, since PI3K/Akt/mTOR has been proposed as a drug target in PEL, we ascertained that pathway-specific inhibitors, namely the dual PI3K and mTOR inhibitor, PF-04691502, and the Akt inhibitor, Akti 1/2, display improved cytotoxicity to PEL cells in hypoxic conditions. Unexpectedly, we found that these drugs reduce lactate production/extracellular acidification rate, and, in combination with the glycolysis inhibitor 2-deoxyglucose (2-DG), they shift PEL cells metabolism from aerobic glycolysis towards oxidative respiration. Moreover, the associations possess strong synergistic cytotoxicity towards PEL cells, and thus may reduce adverse reaction in vivo, while displaying very low toxicity to normal lymphocytes. Finally, we showed that the association of 2-DG and PF-04691502 maintains its cytotoxic and proapoptotic effect also in PEL cells co-cultured with human primary mesothelial cells, a condition known to mimic the in vivo environment and to exert a protective and pro-survival action. All together, these results provide a compelling rationale for the clinical development of new therapies for the treatment of PEL, based on combined targeting of glycolytic metabolism and constitutively activated signaling pathways.

2016 - The bone marrow represents an enrichment site of specific T lymphocytes against filamentous fungi [Articolo su rivista]
Vallerini, Daniela; Riva, Giovanni; Barozzi, Patrizia; Forghieri, Fabio; Lagreca, Ivana; Quadrelli, Chiara; Morselli, Monica; Bresciani, Paola; Cuoghi, Angela; Coluccio, Valeria; Maccaferri, Monica; Paolini, Ambra; Colaci, Elisabetta; Marasca, Roberto; Narni, Franco; Latgè, Jean Paul; Romani, Luigina; Comoli, Patrizia; Campioli, Daniele; Trenti, Tommaso; Luppi, Mario; Potenza, Leonardo
abstract

Bone marrow has already been described as an enrichment site for several antigen-specific T lymphocytes, but the presence of mould-specific T cells has never been investigated in the bone marrow. We have previously demonstrated that mould-specific T cells emerge in the peripheral blood of patients with invasive fungal infections (IFI) but tend to become undetectable after disease resolution. In seven patients with a history of IFI, we investigated the presence of mould-specific T cells secreting different cytokines in bone marrow and peripheral blood paired samples. The results showed that the frequencies of mould-specific T cells secreting the protective cytokine IFNI3 are significantly higher in bone marrow (BM) and are mainly represented by CD8+ T lymphocytes with effector phenotype. A putative disappearance of such protective BM responses after myeloablative therapy could contribute to the increased risk of IFI in hematologic patients.

2016 - Variability of Voriconazole Trough Levels in Haematological Patients: Influence of Comedications with cytochrome P450(CYP) Inhibitors and/or with CYP Inhibitors plus CYP Inducers [Articolo su rivista]
Cojutti, Piergiorgio; Candoni, Anna; Forghieri, Fabio; Isola, Miriam; Zannier, Maria Elena; Bigliardi, Sara; Luppi, Mario; Fanin, Renato; Pea, Federico
abstract

Voriconazole plasma exposure greatly varies among haematological patients. The purpose of this study was to identify the magnitude of influence of comedications with CYP inhibitors and/or with CYP inhibitors plus CYP inducers on voriconazole trough level (Cmin). Voriconazole Cmin was retrospectively assessed among haematological patients who underwent therapeutic drug monitoring (TDM). Univariate and multivariate linear mixed-effect regression analyses were performed to identify the independent predictors of normalized Cmin. Of the 83 included patients, 35 had comedications with CYP inhibitors (omeprazole or pantoprazole) and 21 with CYP inhibitors (omeprazole or pantoprazole) plus CYP inducers (methylprednisolone, dexamethasone, phenobarbital, rifampin or carbamazepine). Median Cmin value (n = 199) was 2.4 mg/L with a wide range of distribution (&lt;0.2-13.5 mg/L). Median (IQR) normalized voriconazole Cmin value was significantly higher in the presence of CYP inhibitors (4.20 mg/L, 3.23-5.51 mg/L) than either in the absence of interacting cotreatments (2.55 mg/L, 1.54-3.47 mg/L) or in the presence of CYP inhibitors plus CYP inducers (2.16 mg/L, 1.19-3.09 mg/L). The presence of CYP inhibitors was highly significantly associated with Cmin &gt;5.5 mg/L (OR: 23.22, 95% CI: 3.01-179.09, p = 0.003). No significant association emerged when CYP inhibitors were coadministered with CYP inducers (OR: 3.53, 95% CI: 0.36-34.95, p = 0.280). The amount of expected Cmin increase was significantly influenced by both the type and the dose of the administered proton pump inhibitor. The study highlights that the benefit from TDM of voriconazole may be maximal in those patients who are cotreated with CYP inhibitors and/or with CYP inhibitors plus CYP inducers, especially when receiving proton pump inhibitors (PPIs) at very high dosages intravenously.

2015 - Antineoplastic effects of liposomal siRNA treatment targeting BLIMP1/PRDM1 in primary effusion lymphoma [Articolo su rivista]
Riva, Giovanni; Lagreca, Ivana; Mattiolo, Adriana; Belletti, Daniela; Lignitto, Laura; Barozzi, Patrizia; Ruozi, Barbara; Vallerini, Daniela; Quadrelli, Chiara; Corradini, Giorgia; Forghieri, Fabio; Marasca, Roberto; Narni, Franco; Tosi, Giovanni; Forni, Flavio; Vandelli, Maria Angela; Amadori, Alberto; Chieco Bianchi, Luigi; Potenza, Leonardo; Calabro', Maria Luisa; Luppi, Mario
abstract

RNA interference (RNAi) has been suggested to represent a promising therapeutic approach in different disease settings. Primary effusion lymphoma (PEL) is a plasmablastic lymphoma consistently expressing B lymphocyte-induced maturation protein 1 (Blimp-1), a pivotal transcriptional regulator during terminal differentiation of B cells into plasma cells. Here we report, for the first time, that transient knockdown of the BLIMP1 gene (also known as PR Domain Containing 1 with ZNF Domain, or PRDM1) using small interfering RNA (siRNA) delivered by liposomes, induced remarkable killing in PEL cell lines. Furthermore, in a murine model of PEL, significantly prolonged survival was achieved by intraperitoneal treatment with such anti-BLIMP1 lipoplexes, while no vector-induced toxicity was observed. This effective and safe RNAi strategy, based on liposomal siRNA targeting a master transcription factor of post-germinal center B cells, may indeed be a potential treatment against plasmablastic lymphoma

2015 - Combination antifungal therapy for invasive mould diseases in haematologic patients. An update on clinical data [Articolo su rivista]
Candoni, Anna; Aversa, Franco; Busca, Alessandro; Cesaro, Simone; Girmenia, Corrado; Luppi, Mario; Rossi, Giuseppe; Venditti, Adriano; Nosari, Anna Maria; Pagano, Livio
abstract

Invasive mould diseases (IMDs) are often encountered in haematologic patients who undergo chemotherapy or who require allogeneic haematopoietic stem cell transplantation (allo-HSCT), and still represent a challenge for physicians. The availability of antifungals with different targets has set the foundation to improve the outcomes of patients with IMDs and also to develop innovative therapeutic approaches. Among these, using combinations of antifungal drugs is an attractive option for reasons such as the broader spectrum of activity, synergy between compounds with different targets, and a reduced risk of fungal resistance. In addition, in vitro studies and animal models have provided evidence supporting the use of combination strategies. Although no controlled, well-powered, prospective clinical trials are yet available to demonstrate the superiority of combination versus monotherapy, the persistently high mortality rate associated with IMDs has stimulated the use of combinations of antifungal drugs, both in adult and paediatric patients. In this paper, we review the recent published literature on combination therapy for the treatment of IMDs in adult and paediatric haematologic patients.

2015 - Detection of Pneumocystis jirovecii and Aspergillus spp. DNa in bronchoalveolar lavage fluids by commercial real-time PCr assays: comparison with conventional diagnostic tests [Articolo su rivista]
Orsi, Carlotta Francesca; Bettua, Clotilde; Pini, Pietro; Venturelli, Claudia; La Regina, Annunziata; Morace, Giulia; Luppi, Mario; Forghieri, Fabio; Bigliardi, Sara; Luppi, Fabrizio; Codeluppi, Mauro; Girardis, Massimo; Blasi, Elisabetta
abstract

The present study employed two commercial real-time PCR kits, MycAssay™ Pneumocystis (PJ-PCR) and MycAssay™ Aspergillus (ASP-PCR), for the search of fungal DNA on 44 bronchoalveolar lavage (BAL) fluids from patients at risk of invasive fungal disease. Operationally, on the basis of clinical diagnosis and according to the European Organization for Research and Treatment Cancer/Mycoses Study Group (EORTC/MSG) criteria, patients were clustered in 3 groups: a P. jirovecii pneumonia (PCP) group, an invasive aspergillosis (IA) group and a control (CTRL) group, consisting of 8, 10 and 24 patients, respectively. The results were compared to those obtained with conventional diagnostic assays, including BAL culture, galactomannan-ELISA (GM) and immunofluorescence (IF). The PJ-PCR assay returned a sensitivity and specificity of 100% and 94.4%, respectively. The ASP-PCR assay showed a sensitivity and specificity of 80% and 97.1%. When compared to the culture assay, the ASP-PCR showed enhanced sensitivity, and a good level of agreement (kappa = 0.63) was observed between ASP-PCR and GM assays. Overall, our data emphasize the diagnostic usefulness of the two commercial real-time PCR assays, especially in high-risk patients where timing is critical and a low fungal burden may hamper correct and prompt diagnosis by conventional tests.

2015 - Differences among young adults, adults and elderly chronic myeloid leukemia patients [Articolo su rivista]
Castagnetti, Fausto; Gugliotta, G.; Baccarani, M.; Breccia, M.; Specchia, G.; Levato, L.; Abruzzese, E.; Rossi, G.; Iurlo, A.; Martino, B.; Pregno, P.; Stagno, F.; Cuneo, A.; Bonifacio, M.; Gobbi, M.; Russo, D.; Gozzini, A.; Tiribelli, M.; De Vivo, A.; Alimena, G.; Cavo, M.; Martinelli, G.; Pane, F.; Saglio, G.; Rosti, G.; Salvi, F.; Pini, M.; Leoni, P.; Rupoli, S.; Galieni, P.; Bigazzi, C.; Cantore, N.; Palmieri, F.; Albano, F.; Russo Rossi, A.; Rambaldi, A.; Intermesoli, T.; Palandri, F.; Testoni, N.; Luatti, S.; Soverini, S.; Iacobucci, I.; Bochicchio, M. T.; Apolinari, M.; Fogli, M.; Cervello, I.; Capucci, A.; Malagola, M.; Malpignano, A.; Girasoli, M.; Angelucci, E.; Usala, E.; Storti, S.; De Biasi, E.; Tagariello, G.; Sartori, R.; Di Raimondo, F.; Vigneri, P.; Impera, S.; Molica, S.; Lanza, F.; Viganò, C.; Grasso, M.; Rapezzi, D.; Cavazzini, F.; Bosi, A.; Santini, V.; Capalbo, S. F.; Spinosa, G.; Pierri, I.; Bergamaschi, M.; Carella, A. M.; Bacigalupo, A.; De Blasio, A.; Ciccone, F.; Di Renzo, N.; Musolino, C.; Russo, S.; Cortelezzi, A.; Morra, E.; Pungolino, E. M.; Luppi, Mario; Marasca, Roberto; Pogliani, E. M.; Gambacorti Passerini, C.; Luciano, L.; Ferrara, F.; Annunziata, M.; Latte, G.; Noli, D.; Rege Cambrin, G.; Fava, C.; Semenzato, G.; Binotto, G.; Fabbiano, F.; Turri, D.; Siragusa, S.; Caracciolo, C.; Musso, M.; Porretto, F.; Aversa, F.; Crugnola, M.; Cazzola, M.; Orlandi, E.; Falini, B.; Falzetti, F.; Visani, G.; Isidori, A.; Fioritoni, G.; Di Lorenzo, R.; Vallisa, D.; Trabacchi, E.; Petrini, M.; Galimberti, S.; Pizzuti, M.; Zaccaria, A.; Salvucci, M.; Ronco, F.; Ielo, D.; Merli, F.; Avanzini, P.; Tosi, P.; Merli, A.; Musto, P.; De Stefano, V.; Sica, S.; Latagliata, R.; De Fabritiis, P.; Trawiska, M.; Majolino, I.; Pacilli, L.; Ronci, B.; Cedrone, M.; Petti, M. C.; Pisani, F.; Tafuri, A.; Montefusco, E.; Iuliano, F.; Dore, F.; Pardini, S.; Bocchia, M.; Defina, M.; Liberati, A. M.; Luzzi, D.; Boccadoro, M.; Ferrero, D.; Vitolo, U.; Gherlinzoni, F.; Calistri, E.; Fanin, R.; Pizzolo, G.; Meneghini, V.; Rodighiero, F.; D'Emilio, A.
abstract

Background: The incidence of chronic myeloid leukemia (CML) increases with age, but it is unclear how the characteristics of the disease vary with age. In children, where CML is very rare, it presents with more aggressive features, including huge splenomegaly, higher cell count and higher blast cell percentage. Patients and methods: To investigate if after childhood the disease maintains or loses these characteristics of aggressiveness, we analyzed 2784 adult patients, at least 18 years old, registered by GIMEMA CMLWP over a 40-year period. Results: Young adults (YAs: 18-29 years old) significantly differed from adults (30-59 years old) and elderly patients (at least 60 years old) particularly for the frequency of splenomegaly (71%, 63% and 55%, P &lt; 0.001), and the greater spleen size (median value: 4.5, 3.0 and 1.0 cm, P &lt; 0.001). According to the EUTOS score, that is age-independent, high-risk patients were more frequent among YAs, than among adult and elderly patients (18%, 9% and 6%, P &lt; 0.001). In tyrosine kinase inhibitors-treated patients, the rates of complete cytogenetic and major molecular response were lower in YAs, and the probability of transformation was higher (16%, 5% and 7%, P = 0.011). Conclusions: The characteristics of CML or the host response to leukemia differ with age. The knowledge of these differences and of their causes may help to refine the treatment and to improve the outcome.

2015 - Early Education for Early, Integrated Palliative Medicine [Articolo su rivista]
Potenza, Leonardo; Galli, Lisa; Morselli, Monica; Bandieri, Elena; Luppi, Mario
abstract

Early Education for Early, Integrated Palliative Medicine

2015 - Epidemiology and clinical outcome of lower respiratory tract infections by respiratory syncytial virus or parainfluenza virus type 3 in adults receiving treatment for either acute leukemia or severe aplastic anemia: a retrospective single center study [Articolo su rivista]
Bigliardi, Sara; Morselli, Monica; Potenza, Leonardo; Riva, Giovanni; Coluccio, Valeria; Maccaferri, Monica; Paolini, Ambra; Colaci, Elisabetta; Fantuzzi, Valeria; Soci, Francesco; Nasillo, Vincenzo; Messerotti, Andrea; Arletti, Laura; Pioli, Valeria; Lugli, Elisabetta; Gilioli, Andrea; Quadrelli, Chiara; Vallerini, Daniela; Barozzi, Patrizia; Lagreca, Ivana; Marasca, Roberto; Narni, Franco; Franceschini, Erica; Codeluppi, Mauro; Mussini, Cristina; Luppi, Mario; Forghieri, Fabio
abstract

Epidemiology and clinical outcome of lower respiratory tract infections by respiratory syncytial virus or parainfluenza virus type 3 in adults receiving treatment for either acute leukemia or severe aplastic anemia: a retrospective single center study

2015 - Immunologic evidence of a strong association between non-Hodgkin lymphoma and simian virus 40 [Articolo su rivista]
Tognon, Mauro; Luppi, Mario; Corallini, Alfredo; Taronna, Angelo; Barozzi, Patrizia; Rotondo, John Charles; Comar, Manola; Casali, Maria Vittoria; Bovenzi, Massimo; D'Agostino, Antonio; Vinante, Fabrizio; Rigo, Antonella; Ferrarini, Isacco; Barbanti Brodano, Giuseppe; Martini, Fernanda; Mazzoni, Elisa
abstract

Non-Hodgkin lymphoma (NHL), the most common cancer of the lymphatic system, is of unknown etiology. The identification of etiologic factors in the onset of NHL is a key event that could facilitate the prevention and cure of this malignancy. Simian virus 40 (SV40) has been considered an oncogenic agent in the onset/progression of NHL.

2015 - Lenalidomide interferes with tumor-promoting properties of nurse-like cells in chronic lymphocytic leukemia [Articolo su rivista]
Fiorcari, Stefania; Martinelli, Silvia; Bulgarelli, Jenny; Audrito, V; Zucchini, Patrizia; Colaci, Elisabetta; Potenza, Leonardo; Narni, Franco; Luppi, Mario; Deaglio, S; Marasca, Roberto; Maffei, Rossana
abstract

Lenalidomide is an immunomodulatory agent clinically active in chronic lymphocytic leukemia patients. The specific mechanism of action is still undefined, but includes modulation of the microenvironment. In chronic lymphocytic leukemia patients, nurse-like cells differentiate from CD14(+) mononuclear cells and protect chronic lymphocytic leukemia cells from apoptosis. Nurse-like cells resemble M2 macrophages with potent immunosuppressive functions. Here, we examined the effect of lenalidomide on the monocyte/macrophage population in chronic lymphocytic leukemia patients. We found that lenalidomide induces high actin polymerization on CD14(+) monocytes through activation of small GTPases, RhoA, Rac1 and Rap1 that correlated with increased adhesion and impaired monocyte migration in response to CCL2, CCL3 and CXCL12. We observed that lenalidomide increases the number of nurse-like cells that lost the ability to nurture chronic lymphocytic leukemia cells, acquired properties of phagocytosis and promoted T-cell proliferation. Gene expression signature, induced by lenalidomide in nurse-like cells, indicated a reduction of pivotal pro-survival signals for chronic lymphocytic leukemia, such as CCL2, IGF1, CXCL12, HGF1, and supported a modulation towards M1 phenotype with high IL2 and low IL10, IL8 and CD163. Our data provide new insights into the mechanism of action of lenalidomide that mediates a pro-inflammatory switch of nurse-like cells affecting the protective microenvironment generated by chronic lymphocytic leukemia into tissues.

2015 - NPM1 mutations may reveal acute myeloid leukemia in cases otherwise morphologically diagnosed as myelodysplastic syndromes or myelodysplastic/myeloproliferative neoplasms [Articolo su rivista]
Forghieri, Fabio; Paolini, Ambra; Morselli, Monica; Bigliardi, Sara; Bonacorsi, Goretta; Leonardi, Giovanna; Coluccio, Valeria; Maccaferri, Monica; Fantuzzi, Valeria; Faglioni, Laura; Colaci, Elisabetta; Soci, Francesco; Nasillo, Vincenzo; Messerotti, Andrea; Arletti, Laura; Pioli, Valeria; Zucchini, Patrizia; Quadrelli, Chiara; Corradini, Giorgia; Giacobbi, Francesca; Vallerini, Daniela; Riva, Giovanni; Barozzi, Patrizia; Lagreca, Ivana; Marasca, Roberto; Narni, Franco; Mecucci, Cristina; Ottaviani, Emanuela; Martinelli, Giovanni; Falini, Brunangelo; Luppi, Mario; Potenza, Leonardo
abstract

NPM1 mutations may reveal acute myeloid leukemia in cases otherwise morphologically diagnosed as myelodysplastic syndromes or myelodysplastic/myeloproliferative neoplasms.

2015 - Nutlin-3 loaded nanocarriers: Preparation, characterization and in vitro antineoplastic effect against primary effusion lymphoma [Articolo su rivista]
Belletti, Daniela; Tosi, Giovanni; Riva, Giovanni; Lagreca, Ivana; Galliania, M; Luppi, Mario; Vandelli, Maria Angela; Forni, Flavio; Ruozi, Barbara
abstract

In this investigation, Nutlin-3 (Nut3), a novel antitumor drug with low water solubility (<0.1mg/L at 25°C), was loaded into liposomes (Lipo-Nut3), polymeric nanoparticles (NPs-Nut3) and nanoparticles engineered with an antibody direct against Syndecan-1/CD 138 (Syn-NPs-Nut3) to obtain carriers targeted to PEL (primary effusion lymphoma). The physicochemical properties of these carriers were determined. Atomic force microscopy showed that all the particles were well formed and spherical in shape. The presence of the antibody on surface led to a significant increase of mean diameter (280 ± 63 nm), PDI (0.3) and the shift of zeta potential towards neutrality (-1 mV). The entrapment efficiency of Lipo-Nut3, NPs-Nut3 and Syn-NPs-Nut3 was 30, 52 and 29%, and drug loading was 1.4, 4.5 and 2.6%, respectively. By performing cytofluorimetric analyses and bromodeoxyuridine (BrdU) assay, the efficacy of nanocarriers to deliver the antineoplastic drug into a PEL cell line namely BCBL-1 (immortalized body cavity B-cell lymphoma) was investigated. Two days after the treatment with 20 μM of Syn-NPs-Nut3, the cell density decreased at about 60% while the cell viability decreased at 56% only 5 days after transfection, when compared with untreated cells. A cell cycle arrest was observed with a significant decrease of cells in S-phase and increasing of apoptotic cell, if compared with untreated control. These results confirms the potential of nanocarriers approaches to deliver antitumor drug with unfavorable chemico-physical properties. Moreover, this study strongly suggests that Syn-NPs-Nut3 can be a valuable drug carrier system for the treatment of PEL lymphoma.

2015 - Philadelphia chromosome-positive acute lymphoblastic leukemia [Articolo su rivista]
Forghieri, Fabio; Luppi, Mario; Potenza, Leonardo
abstract

Philadelphia chromosome-positive Acute Lymphoblastic Leukemia

2015 - Safety profile of Erwinia asparaginase treatment in adults with newly diagnosed acute lymphoblastic leukemia: a retrospective monocenter study [Articolo su rivista]
Bigliardi, Sara; Morselli, Monica; Potenza, Leonardo; Coluccio, Valeria; Maccaferri, Monica; Paolini, Ambra; Colaci, Elisabetta; Fantuzzi, Valeria; Faglioni, Laura; Soci, Francesco; Nasillo, Vincenzo; Messerotti, Andrea; Pedrazzi, Paola; Marietta, Marco; Luppi, Mario; Forghieri, Fabio
abstract

Safety profile of Erwinia asparaginase treatment in adults with newly diagnosed acute lymphoblastic leukemia: a retrospective monocenter study.

2015 - Targeting neoplastic B cells and harnessing microenvironment: the “double face” of ibrutinib and idelalisib [Articolo su rivista]
Maffei, Rossana; Fiorcari, Stefania; Martinelli, Silvia; Potenza, Leonardo; Luppi, Mario; Marasca, Roberto
abstract

Relapsed or refractory diffuse large B-cell lymphoma (DLBCL) not suitable for high dose chemotherapy with autologous stem cell transplantation (ASCT) has a dismal prognosis and no standard therapy. We designed an Italian multicenter retrospective study aimed at evaluating the safety and efficacy of rituximab plus bendamustine (R–B) as salvage treatment in patients not eligible for ASCT because of age and/or comorbidity or in patients with post-ASCT recurrence. Fifty-five patients with a median age of 76 years were included. The overall response rate was 50%, including 28% complete remission and 22% partial remission. The median overall survival (OS) was 10.8 months. The median progression free survival (PFS) was 8.8 months. Eleven patients are still alive and in complete remission at last follow-up (12–71 months). Toxicity was moderate, mainly grades 1 and 2. R–B showed promising efficacy results with an acceptable toxicity profile and should be further investigated, possibly in combination with novel drugs.

2014 - ABO blood group and risk of peripheral arterial thrombosis in patients with atrial fibrillation: A single center survey [Articolo su rivista]
Franchini, M.; Rossi, C.; Frattini, F.; Crestani, S.; Sissa, C.; Meschieri, M.; Giacomini, I.; Luppi, M.; Bonfanti, C.
abstract

2014 - An unusual case of B-ALL occurring in a patient with acute promyelocytic leukemia in remission after two hematopoietic SCTs: whose are the leukemic cells? [Articolo su rivista]
Bigliardi, S; Morselli, M; Potenza, Leonardo; Bresciani, P; Cuoghi, A; Coluccio, Valeria; Riva, Giovanni; Paolini, Ambra; Fantuzzi, Valeria; Faglioni, Laura; Nasillo, Vincenzo; Messerotti, Andrea; Marasca, Roberto; Narni, Franco; Luppi, Mario; Forghieri, Fabio
abstract

We read with interest the recent article by Shiozaki H, et al. (Bone Marrow Transplant 2014; 49: 102–109) reporting a case of donor cell leukemia (DCL) occurring in a patient who previously underwent cord blood (CB) transplantation for myelodysplastic syndrome and describing the clinical and biological differences between DCL arising after hematopoietic SCT (HSCT) from either CB or BM sources. We would like to comment on these issues, reporting an unusual case of DCL in a patient with a long history of acute promyelocytic leukemia (APL).

2014 - An unusual case of splenomegaly and increased lactate dehydrogenase heralding acute myeloid leukemia with eosinophilia and RUNX1-MECOM fusion transcripts [Articolo su rivista]
Forghieri, Fabio; Bigliardi, Sara; Morselli, Monica; Potenza, Leonardo; Fantuzzi, Valeria; Faglioni, Laura; Nasillo, Vincenzo; Messerotti, Andrea; Paolini, Ambra; Luppi, Mario
abstract

We report the first case of acute myeloid leukemia (AML) with RUNX1-MECOM fusion transcripts, showing marked eosinophilia. A 63-year old man admitted in August 2013, had previously been observed in April 2013, because of persisting homogeneous splenomegaly and increased LDH, which were initially attributed to both minor β-thalassemia and previous acute myocardial infarction. However, based upon the retrospective analysis of clinical features combined with the documentation of both JAK2 V617F and c-KIT D816V mutations at AML diagnosis, an aggressive leukemic transformation with eosinophilia of a previously unrecognized myeloproliferative neoplasm, rather than the occurrence of de novo AML, may be hypothesized.

2014 - Chromosomally Integrated HHV-6 [Capitolo/Saggio]
Luppi, M.; Potenza, L.; Morissette, G.; Flamand, L.
abstract

Herpesviruses are members of a diverse family of viruses that colonize all vertebrates from fish to mammals. Interestingly, certain oncogenic herpesviruses such as the Marek's disease virus can be found integrated at low frequencies in the host's chromosomes. In recent years, the consistent and rather frequent detection (in approximately 1% of the human population) of human herpesvirus 6 (HHV-6) viral DNA integrated into human chromosomes has spurred renewed interest in our understanding of how these viruses infect, replicate, and propagate themselves. In this chapter, we provide a historical perspective on chromosomal integration by HHV-6 and present the current state of knowledge on integration and the possible clinical implications associated with HHV-6 chromosomal integration. © 2014 Elsevier B.V. All rights reserved.

2014 - Classification of HHV-6A and HHV-6B as distinct viruses [Articolo su rivista]
Ablashi, Dharam; Agut, Henri; Alvarez Lafuente, Roberto; Clark, Duncan A; Dewhurst, Stephen; Diluca, Dario; Flamand, Louis; Frenkel, Niza; Gallo, Robert; Gompels, Ursula A; Höllsberg, Per; Jacobson, Steven; Luppi, Mario; Lusso, Paolo; Malnati, Mauro; Medveczky, Peter; Mori, Yasuko; Pellett, Philip E; Pritchett, Joshua C; Yamanishi, Koichi; Yoshikawa, Tetsushi
abstract

Shortly after the discovery of human herpesvirus 6 (HHV-6), two distinct variants, HHV-6A and HHV-6B, were identified. In 2012, the International Committee on Taxonomy of Viruses (ICTV) classified HHV-6A and HHV-6B as separate viruses. This review outlines several of the documented epidemiological, biological, and immunological distinctions between HHV-6A and HHV-6B, which support the ICTV classification. The utilization of virus-specific clinical and laboratory assays for distinguishing HHV-6A and HHV-6B is now required for further classification. For clarity in biological and clinical distinctions between HHV-6A and HHV-6B, scientists and physicians are herein urged, where possible, to differentiate carefully between HHV-6A and HHV-6B in all future publications.

2014 - Drug resistance and BCR-ABL kinase domain mutations in Philadelphia chromosome-positive acute lymphoblastic leukemia from the imatinib to the second-generation tyrosine kinase inhibitor era: The main changes are in the type of mutations, but not in the frequency of mutation involvement [Articolo su rivista]
Soverini, Simona; De Benedittis, Caterina; Papayannidis, Cristina; Paolini, Stefania; Venturi, Claudia; Iacobucci, Ilaria; Luppi, Mario; Bresciani, Paola; Salvucci, Marzia; Russo, Domenico; Sica, Simona; Orlandi, Ester; Intermesoli, Tamara; Gozzini, Antonella; Bonifacio, Massimiliano; Rigolin, Gian Matteo; Pane, Fabrizio; Baccarani, Michele; Cavo, Michele; Martinelli, Giovanni
abstract

Patients with Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL) frequently relapse on imatinib with acquisition of BCR-ABL kinase domain (KD) mutations. To analyze the changes that second-generation tyrosine kinase inhibitors (TKIs) have brought in mutation frequency and type, a database review was undertaken of the results of all the BCR-ABL KD mutation analyses performed in the authors' laboratory from January 2004 to January 2013.

2014 - Endothelin-1 promotes survival and chemoresistance in chronic lymphocytic leukemia B cells through eta receptor [Articolo su rivista]
Maffei, Rossana; Bulgarelli, Jenny; Fiorcari, Stefania; Martinelli, Silvia; Castelli, Ilaria; Valenti, Vanessa; Rossi, Davide; Bonacorsi, Goretta; Zucchini, Patrizia; Potenza, Leonardo; Vallisa, Daniele; Gattei, Valter; Del Poeta, Giovanni; Forconi, Francesco; Gaidano, Gianluca; Narni, Franco; Luppi, Mario; Marasca, Roberto
abstract

The endothelin axis, comprising endothelins (ET-1, ET-2 and ET-3) and their receptors (ET(A)R and ETBR), has emerged as relevant player in tumor growth and metastasis. Here, we investigated the involvement of ET-1/ET(A)R axis in chronic lymphocytic leukemia (CLL). CLL cells expressed higher levels of ET-1 and ETA receptor as compared to normal B cells. ET-1 peptide stimulated phosphoinositide-3-kinase and mitogen-activated protein kinase signaling pathways, improved survival and promoted proliferation of leukemic cells throughout ET(A)R triggering. Moreover, the blockade of ET(A)R by the selective antagonist BQ-123 inhibited the survival advantage acquired by CLL cells in contact with endothelial layers. We also found that blocking ET(A)R via BQ-123 interferes with ERK phosphorylation and CLL pro-survival effect mediated by B-cell receptor (BCR) activation. The pro-apoptotic effect of phosphoinositide-3-kinase δ inhibitor idelalisib and mitogen-activated protein kinase inhibitor PD98059 was decreased by the addition of ET-1 peptide. Then, ET-1 also reduced the cytotoxic effect of fludarabine on CLL cells cultured alone or co-cultured on endothelial layers. ET(A)R blockade by BQ-123 inhibited the ET-1-mediated protection against drug-induced apoptosis. Lastly, higher plasma levels of big ET-1 were detected in patients (n = 151) with unfavourable prognostic factors and shorter time to first treatment. In conclusion, our data describe for the first time a role of ET-1/ET(A)R signaling in CLL pathobiology. ET-1 mediates survival, drug-resistance, and growth signals in CLL cells that can be blocked by ET(A)R inhibition.

2014 - Endothelium-mediated survival of leukemic cells and angiogenesis-related factors are affected by lenalidomide treatment in chronic lymphocytic leukemia [Articolo su rivista]
Maffei, Rossana; Fiorcari, Stefania; Bulgarelli, Jenny; Rizzotto, Lara; Martinelli, Silvia; Rigolin, Gian Matteo; Debbia, Giulia; Castelli, Ilaria; Bonacorsi, Goretta; Santachiara, Rita; Forconi, Francesco; Rossi, Davide; Laurenti, Luca; Palumbo, Giuseppe A.; Vallisa, Daniele; Cuneo, Antonio; Gaidano, Gianluca; Luppi, Mario; Marasca, Roberto
abstract

Lenalidomide is an IMID immunomodulatory agent clinically active in patients with chronic lymphocytic leukemia (CLL). We evaluated the activity of lenalidomide inside an in vitro coculture system of endothelial and CLL cells. Lenalidomide was able to inhibit CLL survival advantage mediated by endothelial contact. Moreover, the marked increase of in vitro angiogenesis determined by CLL-derived conditioned media was reduced by lenalidomide. We also analyzed peripheral blood collected from 27 patients with relapsed or refractory CLL being treated with lenalidomide within a phase II trial. Plasma levels of VEGF and THBS-1 decreased, whereas Ang2 and Ang increased during treatment. Patients who respond to lenalidomide showed a more pronounced decrease of VEGF and bFGF than did patients with stable or progressive disease (p = 0.007 and p = 0.005). Furthermore, lenalidomide reduced circulating endothelial cells and endothelial progenitors by increasing the percentage of apoptotic cells. Conversely, for six matched bone marrow biopsies available before and after treatment, we did not detect any modification in vessel density, suggesting a possible mechanism of vessel normalization rather than regression. In conclusion, our study provides further evidence that the anti-CLL effect of lenalidomide is mediated through the alteration of microenvironmental elements, implying the modulation of several angiogenesis-related factors and disruption of CLL crosstalk with endothelial cells.

2014 - Feedbacks and adaptive capabilities of the PI3K/Akt/mTOR axis in acute myeloid leukemia revealed by pathway selective inhibition and phosphoproteome analysis [Articolo su rivista]
Bertacchini, Jessika; Guida, M; Accordi, B; Mediani, Laura; Martelli, A. M; Barozzi, Patrizia; Petricoin, E; Liotta, L; Milani, G; Giordan, M; Luppi, Mario; Forghieri, Fabio; DE POL, Anto; Cocco, L; Basso, G; Marmiroli, Sandra
abstract

Acute myeloid leukemia (AML) primary cells express high levels of phosphorylated Akt, a master regulator of cellular functions regarded as a promising drug target. By means of reverse phase protein arrays, we examined the response of 80 samples of primary cells from AML patients to selective inhibitors of the phosphatidylinositol 3 kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) axis. We confirm that &gt;60% of the samples analyzed are characterized by high pathway phosphorylation. Unexpectedly, however, we show here that targeting Akt and mTOR with the specific inhibitors Akti 1/2 and Torin1, alone or in combination, result in paradoxical Akt phosphorylation and activation of downstream signaling in 70% of the samples. Indeed, we demonstrate that cropping Akt or mTOR activity can stabilize the Akt/mTOR downstream effectors Forkhead box O and insulin receptor substrate-1, which in turn potentiate signaling through upregulation of the expression/phosphorylation of selected growth factor receptor tyrosine kinases (RTKs). Activation of RTKs in turn reactivates PI3K and downstream signaling, thus overruling the action of the drugs. We finally demonstrate that dual inhibition of Akt and RTKs displays strong synergistic cytotoxic effects in AML cells and downmodulates Akt signaling to a much greater extent than either drug alone, and should therefore be explored in AML clinical setting.

2014 - Genetic PTX3 deficiency and aspergillosis in stem-cell transplantation [Articolo su rivista]
Cunha, Cristina; Aversa, Franco; Lacerda, João F; Busca, Alessandro; Kurzai, Oliver; Grube, Matthias; Löffler, Jürgen; Maertens, Johan A; Bell, Alain S; Inforzato, Antonio; Barbati, Elisa; Almeida, Bruno; Santos e. Sousa, Pedro; Barbui, Anna; Potenza, Leonardo; Caira, Morena; Rodrigues, Fernando; Salvatori, Giovanni; Pagano, Livio; Luppi, Mario; Mantovani, Alberto; Velardi, Andrea; Romani, Luigina; Carvalho, Agostinho
abstract

BACKGROUND: The soluble pattern-recognition receptor known as long pentraxin 3 (PTX3) has a nonredundant role in antifungal immunity. The contribution of single-nucleotide polymorphisms (SNPs) in PTX3 to the development of invasive aspergillosis is unknown. METHODS: We screened an initial cohort of 268 patients undergoing hematopoietic stem-cell transplantation (HSCT) and their donors for PTX3 SNPs modifying the risk of invasive aspergillosis. The analysis was also performed in a multicenter study involving 107 patients with invasive aspergillosis and 223 matched controls. The functional consequences of PTX3 SNPs were investigated in vitro and in lung specimens from transplant recipients. RESULTS: Receipt of a transplant from a donor with a homozygous haplotype (h2/h2) in PTX3 was associated with an increased risk of infection, in both the discovery study (cumulative incidence, 37% vs. 15%; adjusted hazard ratio, 3.08; P=0.003) and the confirmation study (adjusted odds ratio, 2.78; P=0.03), as well as with defective expression of PTX3. Functionally, PTX3 deficiency in h2/h2 neutrophils, presumably due to messenger RNA instability, led to impaired phagocytosis and clearance of the fungus. CONCLUSIONS: Genetic deficiency of PTX3 affects the antifungal capacity of neutrophils and may contribute to the risk of invasive aspergillosis in patients treated with HSCT. (Funded by the European Society of Clinical Microbiology and Infectious Diseases and others.).

2014 - High-dose cytarabine in induction treatment improves the outcome of adult patients younger than age 46 years with acute myeloid leukemia: results of the EORTC-GIMEMA AML-12 trial [Articolo su rivista]
Willemze, Roelof; Suciu, Stefan; Meloni, Giovanna; Labar, Boris; Marie, Jean Pierre; Halkes, Constantijn J. M; Muus, Petra; Mistrik, Martin; Amadori, Sergio; Specchia, Giorgina; Fabbiano, Francesco; Nobile, Francesco; Sborgia, Marco; Camera, Andrea; Selleslag, Dominik L. D; Lefrère, Francois; Magro, Domenico; Sica, Simona; Cantore, Nicola; Beksac, Meral; Berneman, Zwi; Thomas, Xavier; Melillo, Lorella; Guimaraes, Jose E; Leoni, Pietro; Luppi, Mario; Mitra, Maria E; Bron, Dominique; Fillet, Georges; Marijt, Erik W. A; Venditti, Adriano; Hagemeijer, Anne; Mancini, Marco; Jansen, Joop; Cilloni, Daniela; Meert, Liv; Fazi, Paola; Vignetti, Marco; Trisolini, Silvia M; Mandelli, Franco; de Witte, Theo
abstract

Cytarabine plays a pivotal role in the treatment of patients with acute myeloid leukemia (AML). Most centers use 7 to 10 days of cytarabine at a daily dose of 100 to 200 mg/m(2) for remission induction. Consensus has not been reached on the benefit of higher dosages of cytarabine.

2014 - Indagini multidirezionali nella diagnosi di infezione fungina invasiva: ricerca di beta-glucano e rilevazione di profili anticorpali specifici [Abstract in Atti di Convegno]
Pini, P; Ardizzoni, A; Orsi, Cf; Bettua, C; Venturelli, C; Girardis, M; Luppi, M; Codeluppi, M; Peppoloni, S; Posteraro, B; Sanguinetti, M; Saez-Roson, A; Moragues, Md; Blasi, E
abstract

2014 - Long-term molecular remission with persistence of BCR-ABL1-specific cytotoxic T cells following imatinib withdrawal in an elderly patient with Philadelphia-positive ALL [Articolo su rivista]
Riva, Giovanni; Luppi, Mario; Lagreca, Ivana; Barozzi, Patrizia; Quadrelli, Chiara; Vallerini, Daniela; Zanetti, Eleonora; Basso, Sabrina; Forghieri, Fabio; Morselli, Monica; Maccaferri, Monica; Paolini, Ambra; Fantuzzi, Valeria; Messerotti, Andrea; Maffei, Rossana; Iacobucci, Ilaria; Martinelli, Giovanni; Marasca, Roberto; Narni, Franco; Comoli, Patrizia; Potenza, Leonardo
abstract

Long-term molecular remission with persistence of BCR-ABL1-specific cytotoxic T cells following imatinib withdrawal in an elderly patient with Philadelphia-positive ALL

2014 - Results of a multicenter, controlled, randomized clinical trial evaluating the combination of piperacillin/tazobactam and tigecycline in high-risk hematologic patients with cancer with febrile neutropenia [Articolo su rivista]
Bucaneve, Giampaolo; Micozzi, Alessandra; Picardi, Marco; Ballanti, Stelvio; Cascavilla, Nicola; Salutari, Prassede; Specchia, Giorgina; Fanci, Rosa; Luppi, Mario; Cudillo, Laura; Cantaffa, Renato; Milone, Giuseppe; Bocchia, Monica; Martinelli, Giovanni; Offidani, Massimo; Chierichini, Anna; Fabbiano, Francesco; Quarta, Giovanni; Primon, Valeria; Martino, Bruno; Manna, Annunziata; Zuffa, Eliana; Ferrari, Antonella; Gentile, Giuseppe; Foà, Robin; Del Favero, Albano
abstract

Empiric antibiotic monotherapy is considered the standard of treatment for febrile neutropenic patients with cancer, but this approach may be inadequate because of the increasing prevalence of infections caused by multidrug resistant (MDR) bacteria.

2014 - Ruxolitinib for pulmonary extramedullary hematopoiesis in myelofibrosis [Articolo su rivista]
Maccaferri, Monica; Leonardi, Giovanna; Marasca, Roberto; Colaci, Elisabetta; Paolini, Ambra; Soci, Francesco; Forghieri, Fabio; Potenza, Leonardo; Narni, Franco; Luppi, Mario
abstract

Here we report an uncommon case of a patient with MF and pulmonary EMH treated with ruxolitinib

2014 - Splenomegaly in hematological malignancies and portal hypertension [Articolo su rivista]
Manenti, Antonio; Forghieri, Fabio; Colasanto, Dario; Luppi, Mario
abstract

Splenomegaly in hematological malignancies and portal hypertension.

2014 - Use of a high sensitive nanofluidic array for the detection of rare copies of BCR-ABL1 transcript in patients with Philadelphia-positive acute lymphoblastic leukemia in complete response [Articolo su rivista]
Iacobucci, Ilaria; Lonetti, Annalisa; Venturi, Claudia; Ferrari, Anna; Papayannidis, Cristina; Ottaviani, Emanuela; Abbenante, Maria Chiara; Paolini, Stefania; Bresciani, Paola; Potenza, Leonardo; Parisi, Sarah; Cattina, Federica; Soverini, Simona; Russo, Domenico; Luppi, Mario; Martinelli, Giovanni
abstract

Monitoring of minimal residual disease (MRD) by quantification of BCR-ABL1 transcript levels has become a main part of the management of patients with BCR-ABL1-positive acute lymphoblastic leukemia (ALL) in treatment with tyrosine kinase inhibitors (TKIs). The failure to achieve molecular negativity shortly after starting TKI has been demonstrated to be predictive of relapse, suggesting that an accurate measurement of low BCR-ABL1 levels may have a role in preventing hematological relapse. Despite the big efforts made by many European laboratories within the European Study Group, at the time of writing a standardized procedure to quantify and express results is still missing for BCR-ABL1-positive ALL. In this study, in order to detect with high sensitivity low levels of BCR-ABL1 transcripts, we used a new technology and a new molecular approach based on microfluidic digital polymerase chain reaction (dPCR) using Taqman chemistry and we compared obtained results with those generated by the conventional method based on reverse transcriptase PCR reaction (RQ-PCR) for BCR-ABL1 and total ABL1, with TaqMan chemistry and with Applied Biosystems instrument. We demonstrated the dPCR is high-sensitive (able to detect a single copy of BCR-ABL1) and reliable (results are comparable to those obtained by BCR-ABL1 quantification with conventional technology), allowing an accurate monitoring of BCR-ABL1-positive ALL patients in complete remission.

2013 - A SEQUENTIAL USE OF TKI, CHEMOTHERAPY AND TRANSPLANT IS ASSOCIATED WITH HIGH COMPLETE REMISSION RATES, DISEASE-FREE AND OVERALL SURVIVAL IN ADULT PH+ ALL. RESULTS OF THE GIMEMA 0904 PROTOCOL [Abstract in Rivista]
S. Chiaretti, A. Vitale; Piciocchi, Al; Fazi, P; Falzetti, F; Nobile, F; Ferrara, F; Luppi, Mario; Nasa, G. La; Fabritiis, P. de; Tedeschi, A; D’Arco, A. Maria; Fanin, R; Fozza, C; Meloni, G; Negulici, A; Propriis, M. De; Guarini, A; Elia, L; Vignetti, M.
abstract

Background: The clinical scenario of Ph+ ALL, considered the most aggres- sive form of leukemia, has changed profoundly following the introduction of 1st and 2nd generation tyrosine kinase inhibitors (TKI). In adult Ph+ ALL, TKI have been administered as part of the backbone of induction chemotherapy together with conventional drugs or, as in two earlier GIMEMA studies, alone in combination with steroids. The use of TKI has enabled high rates of complete hematological remissions (CHR), also in elderly patients. In the GIMEMA 0904 protocol, adult Ph+ ALL patients (≤60 years) were initially treated in induction and consolidation with Imatinib together with chemotherapy. This combination was associated with unacceptable toxicity and the protocol was amended. Imatinib was given as single agent, in combination with steroids, as induction therapy, while chemotherapy was added as consolidation. Aims: Aim of the study was to verify in adult Ph+ ALL the feasibility and efficacy of a sequential scheme based on an induction phase with Imatinib plus steroids, followed by a consolidation with chemotherapy plus Imatinib and, when applicable, by a transplant procedure. Methods: The steroid pre-phase was started from day -6, to allow central molecular screening, up to day 31. Imatinib (600 mg) was administered from day 1 to day 50. Patients who achieved a CHR received as consolidation therapy a cycle of HAM, without discontinuing Imatinib. HAM+Imatinib was planned also for non-responsive cases, followed by a further cycle of chemotherapy. Eligible patients received an allogeneic or autologous stem cell transplant (allo-SCT, auto-SCT). BCR-ABL1 transcript levels were normalized to the number of ABL1 control gene and expressed as a percentage of ABL1. Results: From July 2007 to April 2010, 51 patients have been enrolled; 23 were males and 28 females. The median age was 45.9 years (range: 16.9- 59.7) and the median WBC 28.0 ¥ 109/L (range: 1.4-597). Thirty-nine patients had a p190 fusion transcript, 7 a p210 and 5 had both. Two patients went offstudy for medical decision and toxicity, respectively. After the steroid pre- phase, 38 patients (79%) had a blast reduction ≥75%. At the end of induction (day 50), 47 patients (96%) had achieved a CHR, 1 had a partial response and 1 did not respond. After HAM, also the latter 2 cases obtained a CHR. No deaths in induction were recorded. Of the 43 patients who received HAM, 23 underwent a SCT procedure (20 allo-SCT, 3 auto-SCT), while 20 did not: so far, 3 relapses have occurred in the transplanted group (13%) and 8 in the non-transplanted group (40%). Disease-free survival (DFS) and overall survival (OS) at 36 months are 50.5% and 69.1%, respectively. SCT had no significant impact on DFS, while a trend was observed for overall survival OS (P=0.06). BCR-ABL1 transcript levels decreased during induction therapy, with a highly significant reduction (P<0.0001) between the onset and the end of induction. A further, non-significant, reduction was induced by HAM. Interestingly, a log reduction >1.3 at day 50 (end of induction) was associated with an improved DFS (P=0.03) and a decreased cumulative incidence of relapse (P=0.004). Summary and Conclusions: A sequential approach with Imatinib alone in induction, consolidated by chemotherapy plus Imatinib and followed by a SCT, is a feasible strategy for the management of adult Ph+ ALL patients. We confirm the very high CHR rates following induction with a TKI as single agent with steroids and no deaths in induction. This sequential therapeutic approach is associated with promising DFS and OS rates.

2013 - A hematology consensus agreement on antifungal strategies for neutropenic patients with hematological malignancies and stem cell transplant recipients. [Articolo su rivista]
Girmenia, C; Aversa, F; Busca, A; Candoni, A; Cesaro, S; Luppi, Mario; Pagano, L; Rossi, G; Venditti, A; Nosari, Am
abstract

In the attempt to establish key therapy definitions and provide shared approaches to invasive fungal diseases in neutropenic patients, trials of empiric, preeemptive and targeted antifungal therapy (EAT, PAT and TAT) were reviewed, and a Consensus Development Conference Project was convened. The Expert-Panel concurred that all antifungal treatments, including EAT, should always follow an adequate diagnostic strategy and that the standard definition of PAT may be misleading: being PAT guided by the results of a diagnostic work-up, it should better be termed diagnostic-driven antifungal therapy (DDAT). The Expert-Panel agreed that radiological findings alone are insufficient for the choice of a TAT and that the identification of the etiologic pathogen is needed. The Consensus Agreement proceeded identifying which clinical and microbiological findings were sufficient to start a DDAT and which were not. Finally, an algorithm to rationalize the choice of antifungal drugs on the basis of clinical manifestations, antifungal prophylaxis, instrumental and laboratory findings was drawn up.

2013 - ABO blood group and risk of coronary artery disease [Articolo su rivista]
Franchini, M.; Rossi, C.; Mengoli, C.; Frattini, F.; Crestani, S.; Giacomini, I.; Luppi, M.; Bonfanti, C.
abstract

2013 - Characterization of Specific Immune Responses to Different Aspergillus Antigens during the Course of Invasive Aspergillosis in Hematologic Patients [Articolo su rivista]
Potenza, Leonardo; Vallerini, Daniela; Barozzi, Patrizia; Riva, Giovanni; Forghieri, Fabio; Beauvais, Anne; Beau, Remi; Candoni, Anna; Maertens, Johan; Rossi, Giulio; Morselli, Monica; Zanetti, Eleonora; Quadrelli, Chiara; Codeluppi, Mauro; Guaraldi, Giovanni; Pagano, Livio; Caira, Morena; DEL GIOVANE, Cinzia; Maccaferri, Monica; Stefani, Alessandro; Morandi, Uliano; Tazzioli, Giovanni; Girardis, Massimo; Delia, Mario; Specchia, Giorgina; Longo, Giuseppe; Marasca, Roberto; Narni, Franco; Merli, Francesco; Imovilli, Annalisa; Apolone, Giovanni; Carvalho, Agostinho; Comoli, Patrizia; Romani, Luigina; Latgè, Jean Paul; Luppi, Mario
abstract

Several studies in mouse model of invasive aspergillosis (IA) and in healthy donors have shown that different Aspergillus antigens may stimulate different adaptive immune responses. However, the occurrence of Aspergillus-specific T cells have not yet been reported in patients with the disease. In patients with IA, we have investigated during the infection: a) whether and how specific T-cell responses to different Aspergillus antigens occur and develop; b) which antigens elicit the highest frequencies of protective immune responses and, c) whether such protective T cells could be expanded ex-vivo. Forty hematologic patients have been studied, including 22 patients with IA and 18 controls. Specific T cells producing IL-10, IFN-γ, IL-4 and IL-17A have been characterized through enzyme linked immunospot and cytokine secretion assays on 88 peripheral blood (PB) samples, by using the following recombinant antigens: GEL1p, CRF1p, PEP1p, SOD1p, α1-3glucan, β1-3glucan, galactomannan. Specific T cells were expanded through short term culture. Aspergillus-specific T cells producing non-protective interleukin-10 (IL-10) and protective interferon-gamma (IFN-γ) have been detected to all the antigens only in IA patients. Lower numbers of specific T cells producing IL-4 and IL-17A have also been shown. Protective T cells targeted predominantly Aspergillus cell wall antigens, tended to increase during the IA course and to be associated with a better clinical outcome. Aspergillus-specific T cells could be successfully generated from the PB of 8 out of 8 patients with IA and included cytotoxic subsets able to lyse Aspergillus hyphae. Aspergillus specific T-cell responses contribute to the clearance of the pathogen in immunosuppressed patients with IA and Aspergillus cell wall antigens are those mainly targeted by protective immune responses. Cytotoxic specific T cells can be expanded from immunosuppressed patients even during the infection by using the above mentioned antigens. These findings may be exploited for immunotherapeutic purposes in patients with IA. © 2013 Potenza et al.

2013 - Clinical heterogeneity of de novo 11q deletion chronic lymphocytic leukaemia: prognostic relevance of extent of 11q deleted nuclei inside leukemic clone. [Articolo su rivista]
Marasca, Roberto; Maffei, Rossana; Martinelli, Silvia; Fiorcari, Stefania; Bulgarelli, Jenny; Debbia, Giulia; Rossi, D; Rossi, Fm; Rigolin, Gm; Martinelli, S; Gattei, V; Del Poeta, G; Laurenti, L; Forconi, F; Montillo, M; Gaidano, G; Luppi, Mario
abstract

Deletion on the long arm of chromosome 11 occurs in 5-20% of chronic lymphocytic leukaemia (CLL) patients. We analysed clinical-biological characteristics of 131 CLL patients carrying 11q deletion documented before therapy (de novo 11q deleted CLL). De novo 11q deleted CLL were characterized by high frequencies of unmutated immunoglobulin variable heavy genes, multiple fluorescence in situ hybridization aberrations and lymph node involvement. Factors significantly associated with shorter time to first treatment (TTFT) were advanced Binet stages, high white blood cell count, increased β(2) -microglobulin levels, 17p in addition, splenomegaly and more extensive lymphadenopathy. We found that patients with <25% 11q deleted nuclei (n = 22) experienced longer TTFT compared with patients with ≥25% 11q deleted nuclei (n = 87; median TTFT, 40 vs. 14 months, p = 0.011) and also showed better response to treatments (complete response, 50% vs. 21%, p = 0.016). The variables identified by multivariate analysis as independently associated with reduced TTFT were advanced Binet stages [hazard ratio (HR) 4.69; p < 0.001] and ≥25% 11q deleted nuclei (HR 4.73; p = 0.004). De novo 11q deleted CLLs exhibit variable clinical outcome. The percentage of deleted nuclei inside leukemic clone should be included in the prognostic definition of therapy-naïve 11q deleted CLL patients.

2013 - DEEP SEQUENCING OF THE BCR-ABL KINASE DOMAIN REVEALS A FREQUENCY OF 35INS INSERTION/TRUNCATION HIGHER THAN EXPECTED [Abstract in Rivista]
Benedittis, C. De; Soverini, S; Polakova, K; Brouckova, A; Castagnetti, F; Gugliotta, G; Palandri, F; Papayannidis, C; Klamova, H; Bresciani, P; Coluccio, V; Salvucci, M; Tiribelli, M; Intermesoli, T; Binotto, G; Iacobucci, I; Venturi, C; Luppi, Mario; Ottaviani, E; Bochicchio, M; Cattina, F; Manuela, M; Leo, E; Haferlach, T; Kohlmann, A; Russo, D; Rosti, G; Baccarani, M; Cavo, M; Martinelli, G.
abstract

Background: The spectrum of Bcr-Abl kinase domain mechanisms that con- fer resistance to tyrosine kinase inhibitors (TKIs) in Philadelphia-positive (Ph+) Leukemia is quite heterogeneous. Not always molecular events underlying drug-resistance can be explained by presence of mutations; Bcr-Abl KD inser- tions/deletions can be an alternative mutational mechanisms. The recent devel- opment of “deep-amplicon sequencing” (DS) technologies has opened the way to a more accurate characterization of molecular aberrations in Ph+ Leukemia with higher sensitivity of screening for know and unknown mutations. Aims: We took advantage of a DS approach in order to fully characterize the spectrum of insertions and deletions in CML and Ph+ ALL patients who had developed resistance to one or multiple lines of TKI therapy. Methods: We set up a Bcr-Abl KD mutation screening assay on the Roche GS Junior instrument that allows to reliably detect sequence variants and deletions or insertions with a lower detection limit of 1%. A total of 67 samples from 26 CML and 13 Ph+ ALL patients who had developed resistance to one or multi- ple TKIs (Imatinib, Dasatinib, Nilotinib) were selected for this analysis. In order to reconstruct the dynamics of growth of mutations we evaluated their presence in a serial follow-up samples collected during TKI therapy in 6 patients. Results: DS revealed a 35-base insertion (35INS) in 18/26 (69%) CML and 11/13 (84%) ALL Ph+ patients with an abundance from 1% up to 96% of all Bcr- Abl transcripts. Interestingly DS highlighted an increased expression of 35INS over time in 6 patients (growth ranged from 2% to 96% within a few months). This insertion is known to retain a stop codon which causes the loss of 653 C- terminal amino acids of Bcr-Abl resulting in early termination and a truncated Bcr-Abl1 protein missing a significant portion of the C-terminal regulatory regions. In addition DS detected 2 in-frame deletions in 3 samples, with an abundance from 2% to 19% . This not previously described variants include a 72-nt deletion (1233-1304) between the junction of Abl exon 6 and 7 that caus- es the loss of 24 amino acids (aa 359-383) and a 42-nt deletion in exon 7 (1258-1299) which leads to loss of 14 amino acids (aa 371-384). Summary / Conclusion: Our results show that DS technologies on the GS Junior instrument allow a more accurate characterization of mutational status of patients in comparison to conventional sequencing methods. The higher sensitivity of DS approach allowed to highlight, both in CML and in Ph ALL+ patients, a frequency of 35INS higher than previously reported (60%). The 35INS thus seems to be very frequent in CML and Ph+ ALL patients who devel- op resistance to one or multiple lines of TKI therapies but its abundance is dynamic in individual patients and seems not to be related to TKI therapy. In line with our results, recent 35INS in vitro studies have demonstrated that this insertion is kinase-inactive and should not contribute to TKI-resistance. Although this insertion does not predict for a specific TKIs-resistance its role in Ph+ Leukemia merit additional studies and further analysis of a larger number of samples will be needed to better understand its biological and clinical rele- vance.

2013 - DYNAMICS OF EXPANSION OF TYROSINE KINASE INHIBITOR-RESISTANT MUTANTS AS ASSESSED BY DEEP SEQUENCING OF THE BCR-ABL KINASE DOMAIN: IMPLICATIONS FOR ROUTINE MUTATION TESTING [Abstract in Atti di Convegno]
Soverini, S; Benedittis, C. De; Polakova, K; Brouckova, A; Papayannidis, C; Abbenante, M; Iacobucci, I; Venturi, C; Ferrari, A; Cattina, F; Russo, D; Luppi, Mario; Bresciani, P; Vitale, A; Foà, R; Baccarani, M; Cavo, M; Martinelli, G.
abstract

Background: In Philadelphia-positive (Ph+) acute lymphoblastic leukemia (ALL) patients (pts), efficacy of tyrosine kinase inhibitor (TKI)-based therapies is often compromised by selection of resistant mutations in the BCR-ABL kinase domain (KD). Currently, the gold standard for BCR-ABL KD mutation screen- ing is conventional Sanger sequencing (SS). However, more sensitive approaches are desirable to allow more timely and rational therapeutic intervention. Aims: A Deep sequencing (DS) strategy based on the Roche 454 next-generation sequencing technology was set up in order to: study the dynamics of expansion of different types of BCR-ABL KD mutations in Ph+ ALL patients developing resistance to TKI-based therapies; test the ability of DS to highlight emerging clones harboring TKI-resistant mutations. Methods: 29 Ph+ ALL pts who had developed resistance to TKI-based (imatinib, dasatinib, nilotinib) therapies were selected for this retrospective analysis. All the pts were known to have developed TKI-resistant BCR-ABL mutations on treatment, as assessed by SS. To reconstruct the dynamics of mutation emergence, longitudinal re-analysis of samples from relapse backwards (n=97; 1-3 months sampling interval) was performed on a Roche GS Junior instru- ment. DS runs were designed so as to enable high sensitivity mutation calling (minimum target sequence coverage 4,000 reads). However, to minimize the likelihood of false positive results, data were analyzed filtering out all variants with &lt;1% abundance. Results: DS could successfully detect all the mutations (n=85) previously identified by SS (&gt;15% abundance). In addition, DS revealed that both those samples that had been scored as apparently wild-type by SS and those samples already known to harbor mutations as assessed by SS might be carrying one or more ‘lower level’ mutations (&lt;15% abundance). In the latter cases, clonal analysis showed complex textures with the same mutation alone and also in combination with other(s) (‘compound’ mutations) in distinct subclones. Some lower level mutations were silent or apparently irrelevant from a clinical standpoint (passenger mutations?). In more than half of the cases, however, known TKI-resistant variants could be recognized that corresponded either to ‘withdrawing’ mutants not (yet) entirely de-selected by the switch in TKI or to outgrowing mutations anticipating an imminent relapse. Lower level mutations were confirmed with independent methods (ASO-PCR, RFLP). Notably, in 16/29 (55%) pts with molecularly detectable disease but not yet evidence of cytogenetic or hematologic relapse, DS could identify emerging mutations 1 to 3 months before they became detectable by SS. In the remaining 13 pts, however, outgrowth of the TKI-resistant mutation (T315I=7, Y253H=2, E255K=2, E255V=1 and F317L=1) was so rapid that not even a strict monthly monitoring could have allowed to pick them up before they became dominant. Summary / Conclusion: Now that multiple options are available, BCR-ABL KD mutation monitoring is a precious tool to maximize the efficacy of TKI-based regimens as induction or salvage therapy of Ph+ ALL. DS proved as reliable as SS for the detection of mutations with &gt;15% abundance. As a key advantage, DS added precious quantitative and qualitative information on the full repertoire of mutated populations, that SS underestimated in more than half of the samples analyzed. TKI-resistant mutations leading to patient relapse were not necessarily preexisting at diagnosis or at the time of switchover to another TKI, underlining the importance of regular monitoring of pts. Although the majority of mutations were found to arise and take over very rapidly, a monthly monitoring by our DS approach would have allowed to identify them earlier than SS actually did - and well in advance of clinical relapse - in half of the pts. DS techn

2013 - Early palliative and supportive care in hematology wards. [Articolo su rivista]
Bandieri, E; Apolone, G; Luppi, Mario
abstract

Early palliative and supportive care in hematology wards

2013 - Human Herpesvirus 8 (HHV8) Infection and Related Diseases in Italian Transplant Cohorts. [Articolo su rivista]
Riva, Giovanni; Barozzi, Patrizia; Quadrelli, Chiara; Vallerini, Daniela; Zanetti, Eleonora; Forghieri, Fabio; Chiereghin, A; Libri, I; Maggiore, U; Buzio, C; Lazzarotto, T; Narni, Franco; Luppi, Mario; Potenza, Leonardo
abstract

Human Herpesvirus 8 (HHV8) Infection and Related Diseases in Italian Transplant Cohorts

2013 - Immunity to Polyomavirus BK Infection: Immune Monitoring to Regulate the Balance between Risk of BKV Nephropathy and Induction of Alloimmunity. [Articolo su rivista]
Comoli, P; Cioni, M; Basso, S; Gagliardone, C; Potenza, Leonardo; Verrina, E; Luppi, Mario; Zecca, M; Ghiggeri, Gm; Ginevri, F.
abstract

Polyomavirus BK-associated nephropathy (PyVAN) is the main infectious cause of allograft damage after kidney transplantation. A number of studies revealed an association between the presence of BKV-specific cellular immunity and BK viral clearance, with patients failing to recover specific T cells progressing to PyVAN. Evolution to allograft dysfunction can be prevented by restoration of BKV-specific immunity through a stepwise reduction of maintenance immunosuppressive drugs. Prospective monitoring of BK viral load and specific immunity, together with B-cell alloimmune surveillance, may allow a targeted modification/reduction of immunosuppression, with the aim of obtaining viral clearance while preventing graft injury due to deposition of de novo donor-specific HLA antibodies and late/chronic antibody-mediated allograft injury. Innovative, immune-based therapies may further contribute to BKV infection prevention and control.

2013 - Monocytic population in chronic lymphocytic leukemia shows altered composition and deregulation of genes involved in phagocytosis and inflammation [Articolo su rivista]
Maffei, Rossana; Bulgarelli, Jenny; Fiorcari, Stefania; Bertoncelli, L; Martinelli, Silvia; Guarnotta, C; Castelli, Ilaria; Deaglio, S; Debbia, Giulia; DE BIASI, Sara; Bonacorsi, G; Zucchini, Patrizia; Narni, Franco; Tripodo, C; Luppi, Mario; Cossarizza, Andrea; Marasca, Roberto
abstract

Macrophages reside in tissues infiltrated by chronic lymphocytic leukemia B-cells and the extent of infiltration is associated with adverse prognostic factors. Blood monocyte population was studied by flow cytometry and whole-genome microarrays. A mixed lymphocyte reaction was performed to evaluate T cell proliferation in contact with monocytes from patients and normal donors. Migration and gene modulation in normal monocytes treated with leukemia were also evaluated. Chronic lymphocytic leukemia patients showed an increase in the absolute number of monocytes compared to normal controls (792+/-86 cells/mL vs. 485+/-46 cells/mL, p=0.003). Higher number of nonclassical CD14+CD16++ and Tie-2 expressing monocytes (TEMs) was also detected in patients. Furthermore, we performed a gene expression analysis of monocytes in chronic lymphocytic leukemia patients, showing up-regulation of RAP1GAP and down-regulation of tubulins and CDC42EP3, which would be expected to result in impairment in phagocytosis. We also detected gene alterations such as the down-regulation of PTGR2, a reductase able to inactivate the prostaglandin E2, indicating an immunosuppressive activity. Accordingly, T cell proliferation was inhibited in contact with monocytes from patients compared to normal controls. Finally, normal monocytes in vitro increased migration and up-regulated CD16, RAP1GAP, IL-10, IL-8, MMP9 and down-regulated PTGR2 in response to leukemic cells or conditioned media. In conclusion, altered composition and deregulation of genes involved in phagocytosis and inflammation were found in blood monocytes obtained from chronic lymphocytic leukemia patients, suggesting that leukemia-mediated 'education' of immune elements may also include the establishment of a skewed phenotype in monocyte/macrophage population.

2013 - O blood group and the risk of major bleeding: A single-center survey [Articolo su rivista]
Franchini, M.; Crestani, S.; Rossi, C.; Frattini, F.; Mengoli, C.; Giacomini, I.; Luppi, M.; Bonfanti, C.
abstract

2013 - O blood group is a risk factor for severe mucosal hemorrhage in orally anticoagulated patients [Articolo su rivista]
Franchini, M.; Rossi, C.; Mengoli, C.; Meschieri, M.; Frattini, F.; Crestani, S.; Giacomini, I.; Luppi, M.; Bonfanti, C.
abstract

2013 - Pain and emozional distress in hematological patients throughout all phases of disease: results from a multidisciplinary research team in Modena University Hospital [Abstract in Rivista]
Alfieri, P; Bandieri, E; Berti, A; Bulgarelli, C; Rizzello, F; Favale, V; Forghieri, Fabio; Galli, L; Morselli, M; Potenza, Leonardo; Zanin, R; Artioli, F; Narni, Franco; Luppi, Mario
abstract

Background: According to some outdated reports, physical pain has been considered for many years a rare feature in the majority of blood malignancies, especially in acute leukemias, with the exception of advanced and terminal phases of disease. Unlike myeloma and lymphoma, there are few published data regarding the frequency of pain in patients with leukemia. Based on the modern concept of total cancer pain and on the importance of patient-reported outcomes, routine symptom assessment for hematologic patients should include, together with pain, even emotional distress, expressed in terms of anxiety and depression. Aims: In order to investigate prevalence and clinical relevance of pain and emotional distress in patients with acute myeloid (AML) and lymphoid (ALL) leukemia referred to our center, a multidisciplinary team consisting of nurses, physicians and psychologists has adopted two validated tools in the daily clinical practice: NRS (Numeral Rating Scale) and HADS (Hospital Anxiety and Depression Scale). ESAS (Edmonton Symptom Assessment System) has been compared with HADS with the aim to evaluate its diagnostic accuracy. Methods: NRS, HADS and ESAS scales were administered to newly diagnosed AML and ALL patients at diagnosis (T0), during the neutropenic phase (T15) and at discharge (T30), throughout hospital admissions and different phases of treatment. According to NRS scale pain intensity was classified as absent (0), mild (1-3), moderate (4-6) or severe (7-10). HADS is 14-item scale given by 7 ques- tions related to anxiety and 7 to depression, determining a score from 0 to 21. ESAS is a multiple-item visual analogue scale from 0 to 10. Anxiety and depression were considered positive with HADS 8 and ESAS 2 or more. Sensitivity and specificity tests were also performed. Results were mainly focused on induction phase, bone marrow transplation (BMT) and home care. Results: From June 2007 to December 2011 137 patients with AML and ALL were enrolled in the study (AML=109, ALL=28, M=85, F=52, median age=60). Another cohort of 31 patients referred to the home care program and affected by several blood disorders (NHL=8, MM=6, AML=5, MF=3, ITP=2, MDS=2, AA=1, ALL=1, CLL=1, CML=1, ET=1) was evaluated in parallel (M=18, F=13, median age=79) on monthly basis. 842 questionnaires were collected in the AML-ALL group. At diagnosis pain was reported in 46.2% of cases (mild=30.3%, moderate-severe=15.9%). The highest prevalence and intensi- ty of pain was observed in post-BMT neutropenic phase associated to mucositis (overall pain=61.5%, severe=30.8%). At diagnosis anxiety scores were positive in 33.6% for HADS and 51.1% for ESAS, while depression was present in 22.4% and 42.4% of cases, respectively. A higher prevalence of anxiety and depression was documented at T15 both in induction and post-BMT phases. Considering all HADS questionnaires anxiety and depression were positive in 26.7% and 25.2% of cases, respectively (10% with HADS from 11 upward, accounting for more severe symptoms), while an ESAS score of 2 or more was reported in 36.5% (anxiety) and 31.9% (depression) of cases. In the home care group pain was reported in 78 of all 157 questionnaires (overall pain=49.7%, mild=26.1%, moderate-severe=23.6%). Anxiety and depression were positive in 31.2% and 45.2% of HADS and in 45.9% and 49% of ESAS questionnaires, respectively. Overall test accuracy of ESAS (score of 2 or more) was 77.5% for anxiety and 76.4% for depression. Summary and Conclusions: Pain, anxiety and depression are common symptoms in a significant proportion of acute leukemia patients, impacting quality of life, clinical decisions and outcomes. Compared with HADS, ESAS showed adequate diagnostic accuracy in screening for anxiety and depression, and could be an excellent candidate for large-scale and routine assessment of physical pain, emotional distress and other symptoms, at diagnosis and during the ac

2013 - Retinoic acid and arsenic trioxide for acute promyelocytic leukemia. [Articolo su rivista]
Lo Coco, F; Avvisati, G; Vignetti, M; Thiede, C; Orlando, Sm; Iacobelli, S; Ferrara, F; Fazi, P; Cicconi, L; Di Bona, E; Specchia, G; Sica, S; Divona, M; Levis, A; Fiedler, W; Cerqui, E; Breccia, M; Fioritoni, G; Salih, Hr; Cazzola, M; Melillo, L; Carella, Am; Brandts, Ch; Morra, E; von Lilienfeld Toal, M; Hertenstein, B; Wattad, M; Lübbert, M; Hänel, M; Schmitz, N; Link, H; Kropp, Mg; Rambaldi, A; La Nasa, G; Luppi, Mario; Ciceri, F; Finizio, O; Venditti, A; Fabbiano, F; Döhner, K; Sauer, M; Ganser, A; Amadori, S; Mandelli, F; Döhner, H; Ehninger, G; Schlenk, Rf; Platzbecker, U; Gruppo Italiano Malattie Ematologiche, Dell'Adulto; German Austrian Acute Myeloid Leukemia Study, Group; Study Alliance, Leukemia
abstract

BACKGROUND: All-trans retinoic acid (ATRA) with chemotherapy is the standard of care for acute promyelocytic leukemia (APL), resulting in cure rates exceeding 80%. Pilot studies of treatment with arsenic trioxide with or without ATRA have shown high efficacy and reduced hematologic toxicity. METHODS: We conducted a phase 3, multicenter trial comparing ATRA plus chemotherapy with ATRA plus arsenic trioxide in patients with APL classified as low-to-intermediate risk (white-cell count, ≤10×10(9) per liter). Patients were randomly assigned to receive either ATRA plus arsenic trioxide for induction and consolidation therapy or standard ATRA-idarubicin induction therapy followed by three cycles of consolidation therapy with ATRA plus chemotherapy and maintenance therapy with low-dose chemotherapy and ATRA. The study was designed as a noninferiority trial to show that the difference between the rates of event-free survival at 2 years in the two groups was not greater than 5%. RESULTS: Complete remission was achieved in all 77 patients in the ATRA-arsenic trioxide group who could be evaluated (100%) and in 75 of 79 patients in the ATRA-chemotherapy group (95%) (P=0.12). The median follow-up was 34.4 months. Two-year event-free survival rates were 97% in the ATRA-arsenic trioxide group and 86% in the ATRA-chemotherapy group (95% confidence interval for the difference, 2 to 22 percentage points; P<0.001 for noninferiority and P=0.02 for superiority of ATRA-arsenic trioxide). Overall survival was also better with ATRA-arsenic trioxide (P=0.02). As compared with ATRA-chemotherapy, ATRA-arsenic trioxide was associated with less hematologic toxicity and fewer infections but with more hepatic toxicity. CONCLUSIONS: ATRA plus arsenic trioxide is at least not inferior and may be superior to ATRA plus chemotherapy in the treatment of patients with low-to-intermediate-risk APL. (Funded by Associazione Italiana contro le Leucemie and others; ClinicalTrials.gov number, NCT00482833.).

2013 - Successful liver transplantation in a patient with splanchnic vein thrombosis and pulmonary embolism due to polycythemia vera with Jak2v617f mutation and heparin-induced thrombocytopenia. [Articolo su rivista]
Biagioni, E; Pedrazzi, Paola; Marietta, M; DI BENEDETTO, Fabrizio; Villa, Erica; Luppi, Mario; Girardis, Massimo
abstract

Heparin-induced thrombocytopenia (HIT) is a rare complication of heparin treatment resulting in a severe acquired thrombophilic condition with an associated mortality of about 10 %. We report the first case of successful urgent liver transplantation (LT) in a patient with end-stage liver disease due to a Budd-Chiari syndrome, portal vein thrombosis and pulmonary embolism due to acquired thrombophilia associated to polycythemia vera carrying JAK2V617F gene mutation and HIT in the acute phase. Lepirudin was used to provide anticoagulation in the LT perioperative period that was performed without haemorrhagic and thrombotic complications despite the donor received heparin during liver explantation.

2013 - Trends in immune cell function assay and donor-specific HLA antibodies in kidney transplantation: A 3-year prospective study [Articolo su rivista]
Libri, I; Gnappi, E; Zanelli, P; Reina, M; Giuliodori, S; Vaglio, A; Palmisano, A; Buzio, C; Riva, Giovanni; Barozzi, Patrizia; Luppi, Mario; Cravedi, P; Maggiore, U.
abstract

The immune cell function assay (ICFA) and de novo anti-donor-specific HLA antibodies (DSA) have been proposed as assays for immune monitoring in renal transplantation, but longitudinal studies examining the modification of both parameters over time and their relation with clinical events are lacking. We prospectively measured longitudinal changes in ICFA and DSA levels in 55 kidney transplant recipients over 3-year follow-up (534 visits) and analyzed their relation with the risk of developing acute rejections or infections. Seven patients (12.7%) developed biopsy-proven acute rejection, and 20 (36.4%) developed viral infections. At 3 years posttransplant, 28% of the patients had developed de novo DSA. ICFA levels peaked at 1-2 months posttransplant (p = 0.005) and leveled off thereafter. They were not associated with the risk of acute rejections, viral infections or development of de novo DSA. Instead, the incidence of de novo DSA was higher in patients who previously had viral infections (adjusted-odds ratio of de novo DSA associated with prior infections: 6.03 [95% CI, 1.64-22.06; p = 0.007]). Our prospective, longitudinal study does not support using ICFA to quantify the immune risk in kidney transplantation. Further studies are needed to confirm the relationship between viral infections and the subsequent development of de novo DSA.

2013 - siRNA-BASED THERAPEUTICS: DELIVERY AND TARGETING TO PEL TUMOR BY USING CATIONIC LIPOSOMES [Abstract in Atti di Convegno]
Belletti, Daniela; Riva, Giovanni; Tosi, Giovanni; Lagreca, Ivana; Barozzi, Patrizia; Adriana, Mattiolo; Elena, Negri; Laura, Lignitto; Luigi Chieco, Bianchi; Forni, Flavio; Luppi, Mario; Vandelli, Maria Angela; Maria Luisa, Calabrò; Ruozi, Barbara
abstract

Aims of this research was to develop a “nanomedicine” approach based on siRNA delivery for the treatment of primary effusion lymphoma (PEL). The therapeutic use of antitumoral siRNA requires the development of specifically designed functional vectors, allowing improve¬ment of siRNA stability after systemic admin¬istration, and enabling targeted delivery directly into the neoplastic cells. In this context, liposomes, and particularly cationic liposomes, appears particu¬larly suitable to generate complexes with highly degradable siRNAs, as well as to specifically deliver siRNAs directly into the cytoplasm of the target tumor cells, where RNA interference processes take place. Generally, the electrostatic interaction between the positively charged lipids and the negatively charged nucleic acids leads to the formation of stable lipoplexes, protecting the cargo against nuclease attack and improving the cellular uptake and activity [1. In this context, we are investigating innovative target strategies to improve the treatment of human herpesvirus 8 (HHV8)-associated primary effusion lymphoma (PEL). Primary effusion lymphoma (PEL) is an aggressive B cell non-Hodgkin’s lymphoma, affecting the serous cavities (such as the pleu¬ral, pericardial and abdominal cavities) and preferentially arising in immunocompromised or elderly patients, typically affected by several comorbidities and organ function impairments. PEL therapy has been revealed to be unsuccessful in the vast majority of patients, who are invariably characterized by a poor prognoses. Recently, small interfering RNAs (siRNAs), able to knock-down viral oncogenic proteins, were shown to induce efficient PEL cell apoptosis in vitro and PEL regressions in mice treated with intracavitary injection of lentiviral vectors expressing siRNA precursors[2. Moving from our promising preliminary results in the field of nanotechnologies[3-4, we are developing different lipid-based nanocarriers (cationic and stealth-cationic liposomes), to deliver specific siRNAs to knock-down novel molecular targets (HHV8-encoded microRNAs, viral oncogenic proteins, or host transcription factors) with relevant functions in PEL pathogenesis [5. We are presently testing the delivery efficiency of these nanocarriers and the antineoplastic activity in vitro and in vivo using different PEL-derived cell lines and a previously established PEL mouse model[6. We performed several preliminary technological experiments aimed at optimizing the operative condition to obtain the efficient liposomes/siRNAs complexes. Chemic-physical properties of both liposomes and lipoplexes were evaluated by exploiting microscopic, spectroscopic and gel electrophoresis techniques. In vitro experiments demonstrated a high transfection efficiency of some of our carriers, which stably protected and efficaciously delivered siRNAs into PEL cells. Preliminary experiments using a mixture of siRNAs targeting a specific cellular gene showed a remarkable dose-dependent apoptosis, measured by annexin-V staining, in lipoplexes-transfected PEL cells. Moreover, the in vivo delivery of these therapeutic siRNAs significantly increased the survival time of treated mice compared with control treatment (log-rank test, lipoplexes vs empty liposomes, p=0.002), indicating that our lipoplexes exerted a significant antineoplastic activity. The empty carriers were not toxic in control mice and did not delay PEL development respect untraeted mice. Our data indicate that our lipoplexes may therefore be considered as the basis for the development of useful short interfering RNA delivery vectors to treat PEL tumor. Moreover, we identified a target gene whose suppression exerts a relevant tumoricidal activity on PEL cells in vitro and in vivo, opening new perspectives for PEL treatment.

2012 - Atraumatic splenic rupture in patients with myelodysplastic syndromes: Report of a case occurred during treatment with 5-azacitidine and review of the literature. [Articolo su rivista]
Forghieri, Fabio; Morselli, M; Leonardi, G; Potenza, Leonardo; Bonacorsi, G; Coluccio, V; Paolini, Ambra; Maccaferri, Monica; Colaci, Elisabetta; Fantuzzi, Valeria; Bigliardi, Sara; Zaldini, Piera; Riva, Giovanni; Barozzi, Patrizia; Leonardi, L; Rossi, A; Marasca, Roberto; Narni, Franco; Luppi, Mario
abstract

No abstract available

2012 - Chronic/relapsing lymphadenopathy associated with HHV-6B infection: a new benign clinico-pathologic entity occurring in immunocompetent individuals [Abstract in Rivista]
Forghieri, Fabio; Potenza, Leonardo; Barozzi, Patrizia; Vallerini, Daniela; Riva, Giovanni; Zanetti, E; Quadrelli, C; Morselli, M; Leonardi, G; Maccaferri, M; Paolini, Ambra; Coluccio, Valeria; Colaci, Elisabetta; Pedrazzi, Letizia; Fantuzzi, Valeria; Bigliardi, Sara; Soci, Francesco; Bonacorsi, G; Zaldini, P; Rossi, G; Milani, M; Rivasi, Francesco; Gennari, W; Pecorari, M; Grottola, Antonella; Tagliazucchi, S; Rumpianesi, F; Mattioli, F; Presutti, Livio; Franzoni, Chiara; Gelmini, Roberta; Saviano, Massimo; Cermelli, Claudio; Marasca, Roberto; Narni, Franco; Luppi, Mario
abstract

Background. HHV-6 DNA sequences were disclosed in lymph node (LN) tis- sues of several patients with lymphoid malignancies, but a direct major role of HHV-6 in lymphoid malignant transformation has so far not been confirmed. In contrast, active HHV-6 infection has been associated to either infectious mononucleosis-like syndrome or acute lymphadenitis occurring in febrilepatients with systemic symptoms, or to Rosai-Dorfman disease in which viral antigens have been detected by immunohistochimical (IHC) analyses in both histiocytes and follicular dendritic cells (FDCs). Methods. We have retrospec- tively analyzed clinical and pathological data of 365 adult patients, consecutive- ly observed at our Institution over a period of 5 years (2006-2010), because of enlarged superficial lymph nodes and subsequently undergoing lymphadenec- tomy. In the benign/reactive cases in which well-recognized etiologies have been excluded, an involvement of HHV-6 active infection or reactivation was investigated by molecular and immunohistochemical examinations. Results. Malignant disorders, namely malignant lymphoproliferative disorders or solid cancer metastases, were found in 227 cases (62%), whereas in 138 cases (38%) benign/reactive pictures were documented on lymph node examination. Among these latter cases, a well-recognized etiology was demonstrated in 84 patients (61%), while in 54 cases (39%), a well-defined non-malignant reactive/infectious cause could not be documented. Immunohistochemical analyses resulted negative for both HHV-6A and HHV-6B in 38 of these latter lymph nodes (70%). In 7 patients (13%), a scattered, scanty and aspecific pos- itivity for HHV-6B late protein was documented in rare interfollicular plasma cells and histiocytes. Surprisingly, in 9 patients (17%), immunohistochemical analyses showed HHV-6B positive staining of FDCs, together with scattered positivity of interfollicular cells. These 9 HIV-negative adult patients (median age 42 years, range 18-76 years), with either localized or generalized LAP, were observed for a median follow-up of 38 months (range 28-166). Of note, six of them presented with recurrent LAP (one to 3 recurrences), without evolving into lymphoma. A common LN histological pattern at presentation showed florid fol- licular hyperplasia with concurrent mild paracortical expansion. Three cases also showed features consistent with PTGC. Constitutional symptoms were absent in all patients. The IHC reactions for both HHV-6A and HHV-6B, per- formed on further control cases, represented by 131 LN tissues from patients with either benign LAP induced by other known etiologies or lymphoma, were invariably negative. Serology was positive for both IgM and IgG with high avid- ity suggesting viral reactivation/reinfection. However, the molecular analyses failed to detect HHV-6 viremias in cell-free-serum samples of all the 9 patients with positive HHV-6B IHC staining, while positivity for HHV-6B DNA was dis- closed by PCR analyses in 7 out of the 7 LN tissues studied. Conclusions. We show for the first time that local reactivation/infection of HHV-6B should be con- sidered among the possible causes of chronic/relapsing benign LAP in immuno- competent individuals. IHC is the method of choice for investigating the pres- ence of HHV-6 infection in such cases. HHV-6B may indirectly modulate and trigger the proliferation of lymphocytes, by locally affecting FDCs and LN microenvironment. FDCs may indeed be involved in presenting HHV-6B anti- gens to other immune cells, mainly cortical B lymphocytes.

2012 - How I treat HHV8/KSHV-related diseases in posttransplant patients. [Articolo su rivista]
Riva, Giovanni; Luppi, Mario; Barozzi, Patrizia; Forghieri, Fabio; Potenza, Leonardo
abstract

Posttransplantation human herpesvirus-8 (HHV8)/Kaposi sarcoma herpesvirus (KSHV) primary infection and/or reactivations are associated with uncommon and sometimes fatal, neoplastic, and non-neoplastic diseases. HHV8-related clinical manifestations notably range from Kaposi sarcoma (KS) to either primary effusion lymphoma or multicentric Castleman disease B-cell malignancies, and from polyclonal HHV8-positive plasmacytic lymphoproliferative disorders to bone marrow failure and peripheral cytopenias, associated or not with hemophagocytic syndromes, and to acute hepatitis syndromes. We reviewed the patient series reported in the literature and summarized clinical management aspects, in terms of diagnosis, follow-up, and treatment. We described typical clinical presentations and histopathologic diagnostic features of these diseases, and we discussed the role of HHV8-specific serologic, molecular, and immunologic assays, particularly focusing on recent data from HHV8-specific T-cell monitoring in posttransplantation KS patients. We finally discussed actual therapeutic options, namely, the reduction or discontinuation of immunosuppressive therapy or the switch from calcineurin inhibitors to mTOR inhibitors, as alternatives to antineoplastic chemotherapy, along with the use of antiherpesvirus agents as prophylactic or therapeutic measures, and treatment with rituximab in posttrans-plantation multicentric Castleman disease patients and non-neoplastic HHV8-associated syndromes.

2012 - Impact of early access to a palliative/supportive care intervention on pain management in patients with cancer. [Articolo su rivista]
Bandieri, E; Sichetti, D; Romero, M; Fanizza, C; Belfiglio, M; Buonaccorso, L; Artioli, F; Campione, F; Tognoni, G; Luppi, Mario
abstract

BACKGROUND: No study has so far addressed whether differences do exist in the management of cancer pain between patients receiving usual care by primary specialists and those receiving early palliative/supportive intervention.PATIENTS AND METHODS: A multicentre cross-sectional study in 32 Italian Hospitals has included 1450 patients, receiving analgesic therapy for cancer pain: 602 with access to primary specialist alone (standard care, SC) and 848 with early access to a palliative/supportive care (ePSC) team, concomitant with primary oncology care.RESULTS: Statistically significant differences in the analgesic drug administration according to care model have been evident: non-opioids were more frequently used in SC (9.5% versus 2%; P &lt; 0.001), while strong opioids in ePSC group (80% versus 63%; P &lt; 0.001). The number of patients with severe pain was lower in ePSC compared with SC group (31% versus 17%; P &lt; 0.001). Results of multivariate analysis have shown that ePSC integrated with primary oncologic care (relative risk 0.69; 95% confidence interval 0.48-0.99; P = 0.045) was an independent factor associated with a 31% reduced risk of suffering from severe pain.CONCLUSIONS: An ePSC team provides the most effective standard of analgesic therapy for cancer pain. A randomized clinical trial is needed to confirm these findings.

2012 - LIPOPLEXES PER LA VEICOLAZIONE ED IL DIREZIONAMENTO DI siRNA AL PEL: VALUTAZIONE DELL’EFFICIENZA DI TRATTAMENTO CON SISTEMI LIPIDICI STABILIZZATI [Relazione in Atti di Convegno]
Belletti, Daniela; Riva, Giovanni; Tosi, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela; Ruozi, Barbara
abstract

Per la stabilizzazione e la somministrazione di siRNAs, sono sempre più studiati vettori non virali, ed in particolare liposomi cationici, capaci di stabilizzare, per interazione elettrostatica, materiale genico generando strutture note come lipoplexes (Lpx). In questo contesto, sono stati formulati e testati Lpx attivi nel trattamento del PEL (Primary Effusion Lymphoma), un linfoma altamente aggressivo e con scarse possibilità terapeutiche. Inizialmente, liposomi allestiti con DOTAP sono stati impiegati per complessare 3 differenti siRNAs anti-BLIMP-1 (fattore trascrizionale fondamentale per mantenere lo stato neoplastico del PEL). I Lpx ottenuti sono stati caratterizzati e ne è stata dimostrata l’elevata capacità trasfettiva sulla linea cellulare BCBL-1, modello di PEL. Per una somministrazione efficace, ed in previsione di studi in vivo, i complessi sono stati stabilizzati all’uptake macrofagico, attraverso modificazioni superficiali con PEG. Gli studi tecnologico-formulativi sono stati condotti sia partendo da liposomi pegilati che introducendo la pegilazione su complessi preformati. La complessazione tra liposomi cationici pegilati (preparati con DOTAP e DSPE-PEG) ed i siRNAs non è risultata efficace nello stabilizzare e trasferire il cargo alle cellule in coltura, presumibilmente a causa dell’ingombro e del tamponamento operato dalla copertura idrofila del liposoma, con conseguente limitata interazione tra il vettore ed il materiale genico. Incoraggianti risultatati sono invece derivati dall’applicazione della strategia “post complexation”, ovvero la pegilazione di Lpx cationici preformati (ottenuti tra liposomi DOTAP e siRNAs) mediante incubazione con micelle di DSPE-PEG. Tali “Post PEG-Lpx” hanno mostrato elevata capacità di stabilizzazione dell’attivo con conservata abilità transfettiva

2012 - Pathogenetic mechanisms of hepatitis C virus-induced B-cell lymphomagenesis. [Articolo su rivista]
Forghieri, Fabio; Luppi, Mario; Barozzi, Patrizia; Maffei, Rossana; Potenza, Leonardo; Narni, Franco; Marasca, Roberto
abstract

Hepatitis C virus (HCV) infection is probably the most common chronic viral infection and affects an estimated 180 million people worldwide, accounting for 3% of the global population. Although the liver is considered to be the primary target, extrahepatic manifestations are well recognized among patients with chronic HCV infection. Epidemiological studies have clearly demonstrated a correlation between chronic HCV infection and occurrence of B-cell non-Hodgkin's lymphomas (B-NHL). The clinical evidence that antiviral therapy has a significant role in the treatment at least of some HCV-associated lymphoproliferative disorders, especially indolent B-NHL, further supports the existence of an etiopathogenetic link. However, the mechanisms exploited by HCV to induce B-cell lymphoproliferation have so far not completely clarified. It is conceivable that different biological mechanisms, namely, chronic antigen stimulation, high-affinity interaction between HCV-E2 protein and its cellular receptors, direct HCV infection of B-cells, and "hit and run" transforming events, may be combined themselves and cooperate in a multifactorial model of HCV-associated lymphomagenesis.

2012 - Physical contact with endothelial cells through β1- and β2- integrins rescues chronic lymphocytic leukemia from spontaneous and drug-induced apoptosis and induces a peculiar gene expression profile on leukemic cells. [Articolo su rivista]
Maffei, Rossana; Fiorcari, Stefania; Bulgarelli, Jenny; Martinelli, Silvia; Castelli, Ilaria; Deaglio, S; Debbia, Giulia; Fontana, M; Coluccio, Valeria; Bonacorsi, G; Zucchini, Patrizia; Narni, Franco; Torelli, Giuseppe; Luppi, Mario; Marasca, Roberto
abstract

Background: Chronic lymphocytic leukemia B-cells display prolonged survival in vivo, but when cultured in vitro rapidly undergo spontaneous apoptosis. We hypothesize that interaction with endothelial cells in infiltrated tissues and during recirculation may have a pathogenetic role in chronic lymphocytic leukemia.Design and Methods: We evaluated apoptosis of leukemic cells after co-culture on HUVEC monolayer with addition of Fludarabine and blocking adhesion antibodies. Then, we compared microarray-based expression profiles between leukemic cells at baseline and after co-culture.ùResults: We found that endothelial layer protected leukemic cells from apoptosis inducing a 2-fold mean decrement in apoptotic cells after 2 days co-culture. Moreover, endothelial layer decreased sensitivity of chronic lymphocytic leukemia B-cells to Fludarabine-induced apoptosis. Physical contact with endothelium mediated by both β1- and β2- integrins is essential for survival advantage. In particular, blocking CD106 on endothelial cells or CD18 on leukemic B-cells determined the almost complete abrogation of survival advantage (&gt;70% inhibition of viability). Conversely, a reduction of apoptosis was also measured in leukemic cells cultured in conditioned medium collected after 2 days of co-culture, implying that survival is partially mediated by soluble factors. Overall, the contact with endothelial cells modulated 1,944 genes on chronic lymphocytic leukemia B-cells, establishing a peculiar gene expression profile: up-regulation of angiogenesis-related genes, increase of genes involved in TGFβ and Wnt signalling pathways, secretion of cytokines recruiting stromal cells and macrophages and increase in anti-apoptotic molecules such as Bcl2 and Survivin. Conclusion: Our study supports the notion that endothelial cells are major players in chronic lymphocytic leukemia microenvironment. Adhesion to endothelium strongly sustains survival, protects from drug-induced apoptosis and widely modifies gene expression profile of leukemic cells.

2012 - Severe pneumonia caused by Legionella pneumophila serogroup 11, Italy. [Articolo su rivista]
Grottola, Antonella; Forghieri, Fabio; Meacci, M; Fabio, A; Pozzi, L; Marchegiano, P; Codeluppi, M; Morselli, M; Potenza, Leonardo; Paolini, Ambra; Coluccio, V; Luppi, Mario; Rumpianesi, F; Pecorari, M.
abstract

Legionella pneumophila serogroups (SGs) 1–16 cause pneumonia in humans. Although SG 1 is the serogroup most commonly associated with disease, we report a case of community-acquired legionellosis caused by SG 11.

2012 - Successful generation of p190-BCR ABL-specific T-cell lines for prophylaxis/treatment of minimal residual disease in HSCT recipients with Ph+ acute lymphoblastic leukaemia [Abstract in Rivista]
Basso, S.; Quartuccio, G.; Guido, I.; Quadrelli, C.; Gurrado, A.; Piantoni, L.; Zavras, N.; Cantoni, F.; Falcone, R.; Riva, Giovanni; Barozzi, Patrizia; Potenza, Leonardo; Maccario, R.; Zecca, M.; Luppi, Mario; Comoli, P.
abstract

Recent studies by our groups have demonstrated the presence of BM-homing, BCR-ABL specific cytotoxic T cells (CTL) in Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ALL) patients during sole Imatinib mesylate maintenance treatment, that inversely correlated with minimal residual disease (MRD). This observation supports the notion that anti-tumor T lymphocytes may effectively participate in the control of Ph+ leukemic proliferation, and represents the rationale for a BCR-ABL-targeted cell therapy approach to prophylactically treat leukemic relapse after HSCT in patients with Ph+ALL. The aim of this study was to evaluate the feasibility of expanding BCR-ABL specific CTL from HSCT donors, to be employed as specific DLI after HSCT for Ph+ALL. We conducted scale-up experiments to validate an in vitro culture method to expand BCR-ABL specific CTL from HSCT donors, by peripheral blood mononuclear cell (PBMC) stimulation with 9-20mer peptide pools derived from the p190 BCR-ABL fusion region. T-cell lines, that included a median 70% CD4+ and 29% CD8+ T lymphocytes, were successfully generated from 5/6 individuals. The T-cell lines showed specific INFg production in Elispot assays consistently higher (median 130 SFU/10e5 cells, range 0-198) than non-cultured donor PBMC (median 4 SFU/10e5 cells). In a standard 51chromium release assay, 5 of 6 T-cell lines presented specific cytotoxic activity against PHA blasts pulsed with BCR-ABL peptide mix (median lysis 30%, range 6-65), CD3-redirected activity against P815 cells (median lysis 38%, range 36-52) with minimal residual alloreactivity (median lysis 4%, range 0-15). Our data indicate that BCR-ABL-specific T-cell lines with limited alloreactivity may be expanded from HSCT donors after stimulation with BCR-ABL fusion region-derived peptides. Their efficacy in containment of MRD after allogeneic HSCT for Ph+ALL remains to be evaluated in clinical trials.

2011 - A case of JAK2 V617F-positive myelodysplastic/myeloproliferative neoplasm with unusual morphology, resembling acute promyelocytic leukemia-like disorder with a chronic course. [Articolo su rivista]
Forghieri, Fabio; Morselli, M; Potenza, Leonardo; Maccaferri, Monica; Pedrazzi, Letizia; Coluccio, Valeria; Barozzi, Patrizia; Vallerini, Daniela; Riva, Giovanni; Zanetti, Eleonora; Quarelli, C; Bonacorsi, G; Artusi, Tullio; Zaldini, Piera; Zucchini, Patrizia; Marasca, Roberto; Narni, Franco; Falini, B; Torelli, Giuseppe; Luppi, Mario
abstract

An atypical case of myelodysplastic syndrome with morphologic aspect resempling acute promyelocytic leukemia, carring JA2 mutations.

2011 - BCR-ABL-specific cytotoxic T cells in the bone marrow of patients with Ph(+) acute lymphoblastic leukemia during second-generation tyrosine-kinase inhibitor therapy. [Articolo su rivista]
Riva, Giovanni; Luppi, Mario; Quadrelli, Chiara; Barozzi, Patrizia; Basso, S; Vallerini, Daniela; Zanetti, E; Morselli, M; Forghieri, Fabio; Maccaferri, M; Paolini, Ambra; DEL GIOVANE, Cinzia; D'Amico, Roberto; Marasca, Roberto; Narni, Franco; Iacobucci, I; Martinelli, G; Baccarani, M; Comoli, P; Potenza, Leonardo
abstract

BCR-ABL-specific cytotoxic T cells in the bone marrow of patients with Ph+ acute lymphoblastic leukemia during second-generation tyrosine-kinase inhibitor therapy

2011 - CDKN2A/B alterations impair prognosis in adult BCR-ABL1-positive acute lymphoblastic leukemia patients. [Articolo su rivista]
Iacobucci, I; Ferrari, A; Lonetti, A; Papayannidis, C; Paoloni, F; Trino, S; Storlazzi, Ct; Ottaviani, E; Cattina, F; Impera, L; Abbenante, Mc; Vignetti, M; Vitale, A; Potenza, Leonardo; Paolini, S; Soverini, S; Pane, F; Luppi, Mario; Foà, R; Baccarani, M; Martinelli, G.
abstract

PURPOSE: The 9p21 locus, encoding three important tumor suppressors (p16/CDKN2A, p14/ARF, and p15/CDKN2B), is a major target of inactivation in the pathogenesis of many human tumors.PATIENTS AND METHODS: To explore, at high resolution, the frequency and size of alterations affecting this locus in adult BCR-ABL1-positive acute lymphoblastic leukemia (ALL) and to investigate their prognostic value, 112 patients (101 de novo and 11 relapsed cases) were analyzed by genome-wide single-nucleotide polymorphism arrays and gene candidate deep exon sequencing. Paired diagnosis-relapse samples were further available and analyzed for 19 (19%) cases.RESULTS: CDKN2A/ARF and CDKN2B genomic alterations were identified in 29% and 25% of newly diagnosed patients, respectively. Deletions were monoallelic in 72% of cases, and in 43% of them, the minimal overlapping region of the lost area spanned only the CDKN2A/B gene locus. An analysis conducted at relapse showed an increase in the detection rate of CDKN2A/ARF loss (47%) compared with the time of diagnosis (P = 0.06). Point mutations within the 9p21 locus were found at very low levels, with only a nonsynonymous substitution in the exon 2 of CDKN2A. Of note, deletions of CDKN2A/B were significantly associated with poor outcomes in terms of overall survival (P = 0.0206), disease free-survival (P = 0.0010), and cumulative incidence of relapse (P = 0.0014).CONCLUSIONS: Inactivation of the 9p21 locus by genomic deletion is a frequent event in BCR-ABL1-positive ALL. Deletions are frequently acquired during leukemia progression and are a poor prognostic marker of long-term outcomes.

2011 - Chromosomally integrated human herpesvirus 6: questions and answers. [Articolo su rivista]
Pellett, Pe; Ablashi, Dv; Ambros, Pf; Agut, H; Caserta, Mt; Descamps, V; Flamand, L; Gautheret Dejean, A; Hall, Cb; Kamble, Rt; Kuehl, U; Lassner, D; Lautenschlager, I; Loomis, Ks; Luppi, Mario; Lusso, P; Medveczky, Pg; Montoya, Jg; Mori, Y; Ogata, M; Pritchett, Jc; Rogez, S; Seto, E; Ward, Kn; Yoshikawa, T; Razonable, Rr
abstract

Chromosomally integrated human herpesvirus 6 (ciHHV-6) is a condition in which the complete HHV-6 genome is integrated into the host germ line genome and is vertically transmitted in a Mendelian manner. The condition is found in less than 1% of controls in the USA and UK, but has been found at a somewhat higher prevalence in transplant recipients and other patient populations in several small studies. HHV-6 levels in whole blood that exceed 5.5 log10 copies/ml are strongly suggestive of ciHHV-6. Monitoring DNA load in plasma and serum is unreliable, both for identifying and for monitoring subjects with ciHHV-6 due to cell lysis and release of cellular DNA. High HHV-6 DNA loads associated with ciHHV-6 can lead to erroneous diagnosis of active infection. Transplant recipients with ciHHV-6 may be at increased risk for bacterial infection and graft rejection. ciHHV-6 can be induced to a state of active viral replication in vitro. It is not known whether ciHHV-6 individuals are put at clinical risk by the use of drugs that have been associated with HHV-6 reactivation in vivo or in vitro. Nonetheless, we urge careful observation when use of such drugs is indicated in individuals known to have ciHHV-6. Little is known about whether individuals with ciHHV-6 develop immune tolerance for viral proteins. Further research is needed to determine the role of ciHHV-6 in disease.

2011 - Chronic eosinophilic leukaemia with ETV6-NTRK3 fusion transcript in an elderly patient affected with pancreatic carcinoma [Articolo su rivista]
Forghieri, Fabio; Morselli, M; Potenza, Leonardo; Maccaferri, Monica; Pedrazzi, Letizia; Paolini, Ambra; Bonacorsi, G; Artusi, Tullio; Giacobbi, F; Corradini, G; Barozzi, Patrizia; Zucchini, Patrizia; Marasca, Roberto; Narni, Franco; Crescenzi, B; Mecucci, C; Falini, B; Torelli, Giuseppe; Luppi, Mario
abstract

Chronic eosinophilic leukaemia with ETV6-NTRK3 fusion transcript in an elderly patient affected with pancreatic carcinoma.A case report

2011 - Complessazione di SiRNA anti Blimp-1/PRDM con liposomi cationici: caratterizzazione chimico-fisica e validazione in vitro su cellule di PEL [Relazione in Atti di Convegno]
Belletti, Daniela; Riva, Giovanni; Forni, Flavio; Barozzi, Patrizia; Luppi, Mario; Tosi, Giovanni; Vandelli, Maria Angela; Ruozi, Barbara
abstract

La tecnologia siRNAs (Small Interfering RNAs) è una innovativa strategia di regolazione genica post-trascrizionale potenzialmente applicabile in diversi campi della medicina ed in particolare in campo oncologico. [1] I siRNAs sono piccole sequenze di RNA a doppio filamento che a livello citoplasmatico subiscono un processo di attivazione mediato da un complesso sistema enzimatico denominato RISC (RNA induced silencing complexes) e divengono capaci di riconoscere e “silenziare” l’RNA target. In particolare le nostre ricerche sono indirizzate al linfoma effusivo delle cavità sierose (PEL). Si tratta di un raro tipo di linfoma di tipo non Hodgkin il cui agente eziologico è l’oncovirus HHV-8; nonostante i differenti approcci chemioterapici tentati, ad oggi non esiste un trattamento farmacologico efficace [2]. La disregolazione specifica mediata da siRNA del network di trascrizione della cellula malata potrebbe rappresentare una possibile alternativa nel trattamento della patologia. Considerando l’instabilità in vivo dei siRNAs associata alla loro scarsa capacità di penetrazione cellulare, il successo della strategia di silenziamento è strettamente dipendente dall’utilizzo di sistemi di delivery capaci di proteggere e trasportare selettivamente l’attivo nella cellula malata [3]. I liposomi, ed in particolare quelli cationici, sono sistemi di veicolazione e direzionamento innovativi, studiati e utilizzati da diversi anni, per la loro capacità di stabilizzare e trasferire materiale genico al bersaglio; la modificazione superficiale con molecole stabilizzanti (es PEG) e ligando selettive quali anticorpi monoclonali, peptidi ed aptameri garantisce la veicolazione e selettività del sistema [4,5].In studi preliminari, abbiamo dimostrato come sistemi liposomiali “stealth” modificati con anticorpo anti CD-138 specificamente riconosciuto da proteoglicani largamente espressi sulle cellule linfomatose, risultino altamente efficienti nella complessazione e stabilizzazione di materiale genico. Tali immunoliposomi sono stati testati quali carriers di un oligonucleotide modello (ODN-FITC) su cellule BCBL-1 (linea cellulare di PEL) evidenziando una buona capacità di trasferimento e di targeting valutata mediante citofluorimetria e microscopia confocale [6]. Tali evidenze sono risultate basilari per intraprendere nuovi studi di ottimizzazione di sistemi veicolanti siRNAs specifici nel silenziamento di fattori trascrizionali dominanti dello stadio plasmacellulare (knock-down di BLIMP-1/PRDM) in cellule PEL.In questa ricerca sono stati dapprima formulati e caratterizzati vettori liposomiali (Lp) utilizzando lipidi cationici (Dotap e DC-Chol) e neutri (Dope e Pc). Sono stati così selezionati i vettori che presentavano migliori caratteristiche in termini di dimensioni, carica superficiale e stabilità ed è stata valutata la loro capacità di complessare e stabilizzare siRNAs anti BLIMP-1. I complessi ottimizzati sono stati caratterizzati dal punto di vista chimico fisico mediante PCS e AFM ed è stata valutata l’efficienza di complessazione mediante elettroforesi su gel di agarosio. I dati hanno evidenziato come i liposomi allestiti con il lipide cationico Dotap risultino quelli maggiormente idonei alla formazione di lipoplexes idonei alla somministrazione: caratterizzati da strutture definite e riproducibili, di dimensioni prossime a 350nm che efficientemente proteggono i siRNAs, mostrando inoltre una buona stabilità dopo incubazione in siero. I lipoplexes allestiti con Dotap sono inoltre stati testati in vitro sulla linea cellulare BCBL-1, tali vettori sono risultati atossici in un ampio range di concentrazioni ed abili nel trasferire il materiale genico a livello citoplasmatico. In particolare è stato valutato l’effetto mediato dall’inibizione della proteina bersaglio mediante determinazione della proliferazione cellulare (analisi citomorfologica e conta cellulare) e test annessina/propidio per valutare

2011 - Dasatinib as first-line treatment for adult patients with Philadelphia chromosome-positive acute lymphoblastic leukemia. [Articolo su rivista]
Foà, R; Vitale, A; Vignetti, M; Meloni, G; Guarini, A; De Propris, Ms; Elia, L; Paoloni, F; Fazi, P; Cimino, G; Nobile, F; Ferrara, F; Castagnola, C; Sica, S; Leoni, P; Zuffa, E; Fozza, C; Luppi, Mario; Candoni, A; Iacobucci, I; Soverini, S; Mandelli, F; Martinelli, G; Baccarani, M; GIMEMA Acute Leukemia Working, Party
abstract

Dasatinib is a potent BCR-ABL inhibitor effective in chronic myeloid leukemia and Ph(+) acute lymphoblastic leukemia (ALL) resistant/intolerant to imatinib. In the GIMEMA LAL1205 protocol, patients with newly diagnosed Ph(+) ALL older than 18 years (with no upper age limit) received dasatinib induction therapy for 84 days combined with steroids for the first 32 days and intrathecal chemotherapy. Postremission therapy was free. Fifty-three patients were evaluable (median age, 53.6 years). All patients achieved a complete hematologic remission (CHR), 49 (92.5%) at day 22. At this time point, 10 patients achieved a BCR-ABL reduction to < 10(-3). At 20 months, the overall survival was 69.2% and disease-free survival was 51.1%. A significant difference in DFS was observed between patients who showed at day 22 a decrease in BCR-ABL levels to < 10(-3) compared with patients who never reached these levels during induction. In multivariate analysis, BCR-ABL levels of < 10(-3) at day 85 correlated with disease-free survival. No deaths or relapses occurred during induction. Twenty-three patients relapsed after completing induction. A T315I mutation was detected in 12 of 17 relapsed cases. Treatment was well tolerated; only 4 patients discontinued therapy during the last phase of the induction when already in CHR. In adult Ph(+) ALL, induction treatment with dasatinib plus steroids is associated with a CHR in virtually all patients, irrespective of age, good compliance, no deaths, and a very rapid debulking of the neoplastic clone.

2011 - Differences in Kaposi sarcoma-associated herpesvirus-specific and herpesvirus-non-specific immune responses in classic Kaposi sarcoma cases and matched controls in Sicily. [Articolo su rivista]
Amodio, E; Goedert, Jj; Barozzi, Patrizia; Riva, Giovanni; Firenze, A; Bonura, F; Viviano, E; Romano, N; Luppi, Mario
abstract

Kaposi sarcoma (KS) might develop because of incompetent immune responses, both non-specifically and specifically against the KS-associated herpesvirus (KSHV). Peripheral blood mononuclear cells from 15 classic (non-AIDS) KS cases, 13 KSHV seropositives (without KS) and 15 KSHV-seronegative controls were tested for interferon-γ T-cell (enzyme-linked immunospot [Elispot]) responses to KSHV-latency-associated nuclear antigen (LANA), KSHV-K8.1 and CMV/Epstein-Barr virus (EBV) peptide pools. The forearm and thigh of each participant was also tested for delayed-type hypersensitivity (DTH) against common recall antigens. Groups were compared with Fisher exact test and multinomial logistic regression to calculate odds ratios (OR) and 95% confidence intervals (CI). A KSHV Elispot response was detected in 10 (67%) classic KS cases, 11 (85%) KSHV seropositives (without KS) and two (13%) seronegative controls. All four cases with KSHV-LANA responses had current KS lesions, whereas five of six cases with KSHV-K8.1 responses had no lesions (P = 0.048). No case responded to both LANA and K8.1. Compared with the seronegative controls, the risk for classic KS was inversely related to DTH in the thigh (OR 0.71, 95% CI 0.55-0.94, P = 0.01), directly associated with DTH in the forearm (OR 1.35, 95% CI 1.02-1.80, P = 0.04) and tended to be increased fivefold per KSHV Elispot response (OR 5.13, 95% CI 0.86-30.77, P = 0.07). Compared with KSHV seropositives (without KS), the risk for classic KS was reduced fivefold (OR 0.20, CI 0.03-0.77, P = 0.04) per KSHV response. The CMV/EBV Elispot responses were irrelevant. Deficiency of both KSHV-specific and KSHV-non-specific immunity is associated with classic KS. This might clarify why Kaposi sarcoma responds to immune reconstitution.

2011 - Formulazione di siRNA anti blimp-1/PRDM in liposomi cationici: caratterizzazione chimico-fisica e validazione in vitro su cellule di LINFOMA EFFUSIVO PRIMARIO (pel) [Abstract in Atti di Convegno]
Belletti, Daniela; Riva, Giovanni; Barozzi, Patrizia; Baraldi, Elisa; Veratti, Patrizia; Luppi, Mario; Tosi, Giovanni; Ruozi, Barbara
abstract

Formulazione di siRNA anti blimp-1/PRDM in liposomi cationici: caratterizzazione chimico-fisica e validazione in vitro su cellule di LINFOMA EFFUSIVO PRIMARIO (pel)

2011 - INTERACTION BETWEEN ENDOTHELIUM AND CHRONIC LYMPHOCYTIC LEUKEMIA B-CELLS RESCUES FROM APOPTOSIS AND MODULATES GENE EXPRESSION PROFILE OF LEUKEMIC CELLS [Abstract in Rivista]
Maffei, Rossana; Fiorcari, Stefania; Martinelli, Silvia; Bulgarelli, Jenny; Debbia, Giulia; Fontana, M; Faglioni, Laura; Bigliardi, Sara; Zucchini, Patrizia; Narni, Franco; Torelli, Giuseppe; Luppi, Mario; Marasca, Roberto
abstract

Background. Despite an apparent long life in vivo, CLL cells die rap- idly in vitro. This observation suggests that the apoptotic resistance is not intrinsic to leukemia B cells but extrinsic factors are necessary for CLL prolonged survival. Aims. we investigated the interactions be- tween endothelial cells and CLL cells, highlighting molecular net- works involved in this cellular crosstalk. Methods. we co-cultured CLL cells on HUVEC endothelial monolayer (HC) or in medium alone (CLL only). Then, we detected CLL viability by flow cytometry and we performed whole-genome high density microarrays. Results. we found that endothelial cells protected CLL from spontaneous apop- tosis. After 48h, increased number of alive CLL cells was present in HC condition (59.7 ± 4.2%) compared to CLL alone (22.9 ± 5.1%) (p<0.0001). Moreover, we found that spontaneous in vitro apoptosis was higher in unmutated IGHV CLL (UM-CLL) compared to mutated ones (M-CLL). In HC condition, similar survival was detected be- tween M-CLL and UM-CLL, implying a 2.2-fold increase in relative viability in M-CLL and a 6.1-fold increase in UM-CLL. Moreover, the endothelial cell layer decreased the in vitro sensitivity of CLL cells to Fludarabine-induced apoptotic cell death. The mean viability of CLL cells treated with 10 µM Fludarabine was 19.8% (±4.4%) after 48 hours and 3.8% (±1.3%) after 72 hours. In HC with Fludarabine ad- dition, the mean viability of CLL cells was 37.8% (±9.1%) after 48 hours and 14.3% (±3.2%) after 72 hours. Then, we compared gene expression profiles (GEP) between CLL cultured in contact with EC layer and CLL at baseline to unravel the transcriptional modifications induced by EC cells. Overall 1944 genes were found to be modulated (FC≥2, p<0.05). CLL cells in HC condition showed a 22.6-fold in- crease of CCL2, able to recruit tumor-activated monocytes (p=0.0032) and a 6.5-fold increase of PDGFC, chemoattractant for mesenchymal stromal cells (p=0.0051). Other soluble factors up-reg- ulated by EC/CLL contact were VEGFC (FC=9.4, p=0.0061), ANGTL4 (FC=8.6, p=0.015), EDN1 (FC=9.2, p=0.0061), AMOTL2 (FC=4.3, p=0.019) and THBS1 (FC=45.1, p=0.0004) as well as the metalloproteases MMP2 (FC=8.3, p=0.02) and MMP4 (FC=3.0, p=0.039). The GEP data were confirmed by evaluating the secreted levels of soluble factors in conditioned medium collected after 48h- HC culture. In addition, CLL cells on endothelial layer maintained or increased the expression levels of anti-apoptotic factors Bcl-2, Bcl2A1, BIRC3/c-IAP2 and BIRC5/Survivin compared to CLL cells at baseline. Of interest, the Ang2 tyrosine kinase receptor Tie2 mRNA was found to be increased in CLL cells in co-culture (FC=10.7, p=0.017). We confirmed GEP data by flow cytometry finding a 2-fold and a 4.3-fold increase of percentage of Tie2+CLL cells at 48h and 72h in HC. Conclusion. our results demonstrate a role of endothelial cells in CLL survival advantage and Fludarabine-resistance. The inti- mate contacts with EC seem to determine a microenvironmental- driven angiogenic switch of CLL phenotype, improve the secretion of cytokines involved in regulation of microenvironmental elements such as stromal cells and macrophages and increase the expression of anti-apoptotic molecules.

2011 - Identification and characterization of Aspergillus-specific immune responses to diagnose invasive aspergillosis in high risk patients: a multicenter study [Abstract in Rivista]
Potenza, Leonardo; Vallerini, Daniela; Barozzi, Patrizia; Riva, Giovanni; Maertens, J; Candoni, A; Beauvais, A; Zanetti, Eleonora; Quadrelli, C; Morselli, M; Forghieri, Fabio; Maccaferri, M; Paolini, Ambra; Marasca, Roberto; DEL GIOVANE, Cinzia; D'Amico, Roberto; Ciceri, F; Comoli, P; Cesaro, S; Caira, M; Pagano, L; Romani, L; Narni, Franco; Latgè, Jp; Luppi, Mario
abstract

Background. The mortality of Invasive Aspergillosis (IA) still affects from 27% to 55% of high risk hematologic patients. The reasons of such a poor outcome also rely on several drawbacks limiting the di- agnostic accuracy of non cultural based diagnostic methods (NCBDM) and hampering the opportunities for an early intervention. Studies in mice model of IA and in healthy subjects have shown that Aspergillus-specific T-cells producing interferon-gamma (IFN- gamma-T1) are protective, while Aspergillus-specific T-cells pro- ducing interleukin-10 (IL-10-T2) are non-protective to IA. Aims. We have investigated whether the identification of Aspergillus-specific IFN-gamma-T1 and/or IL-10-T2 through an ex-vivo enzyme linked immunospot (ELISPOT) assay may be effective in the diagnosis of IA in high risk patients. Furthermore, in the proven IA patients, we have functionally and phenotipically characterized such T cells through the cytokine secretion assay (CSA). Methods. 180 patients (168 hemato- logic and 12 solid organ transplant patients) have been enrolled. They were classified, according the revised EORTC/MSG criteria, as fol- lows: 18 proven, 35 probable, 17 possible IA cases and 110 controls. The control patients were divided in two groups: group 1 included 86 (78.2%) patients with hystological and/or cultural verified infec- tious/inflammatory/neoplastic diseases, but other than IA; group 2 in- cluded 24 (21.8%) patients without clinical and/or microbiological features of IA. ELISPOT has been performed, as described [Potenza et al. Leukemia 2007; 21: 578-81], by using as antigens Aspergillus either conidia or recombinant antigens, namely CRF1p, GEL1p, PEP1p, SOD1p, α1-3 glucan, β1-3 glucan and galactomannan (GM). Results. The patient and sample positivity rates were 94.4%/89.5% in proven, 45.7%/35.3% in probable, 35.3%/50% in possible IA cases and 1.8%/4.5% in the controls, respectively. The sensitivity and speci- ficity of ELISPOT for the diagnosis of IA resulted 94.4% (95% CI, 73%-99%) and 98.2% (95% CI, 93%-99%), respectively. The PPV of the test was 89.5% (95% CI, 67%-99%), the NPV was 99.1% (95% CI, 94%-100%) and the efficiency was 97.6% (95% CI, 92.3%- 99.4%). The positive likelihood ratio (LR) resulted 51.89, the negative LR was 0.06 (Table 1A,B). In proven IA patients, CSA demonstrated that Aspergillus-specific IL-10-T2 were predominantly central memory (CM) CD4+ T cells (median frequency 0.37%/0.22%), while Aspergillus-specific IFN-gamma-T1 were either CD4+ or CD8+ cells of either effector memory (EM) or CM phenotype (median frequen- cies 0.24%/0.20%). Also lower frequencies of Aspergillus-specific ei- ther CD4+ or CD8+ T cells producing IL-4 (0.11%/0.19%) of EM phenotype, and EM CD8+ cells producing IL-17 (0.18%), were de- tected. Moreover, although CRF1p, GEL1p, α1-3 glucan and SOD1p resulted the antigens eliciting the highest number of Aspergillus-spe- cific IFN-gamma-T1, only GEL1p and α1-3 glucan were those most constantly targeted by protective immune responses along the entire course of the IA. Conclusions. Our findings demonstrate the potential of ELISPOT in the diagnosis of IA, suggesting that it may comple- ment the other NCBDM, enabling a more consistent diagnosis of IA. Furthermore, this study describes for the first time the Aspergillus- specific immune responses in patients with proven IA, identifying also the antigens predominantly targeted by protective IFN-gamma- T1, with possible consequences in designing strategies of either adoptive cell infusion or vaccine therapies.

2011 - KNOCK-DOWN DI BLIMP1/PRDM1 IN CELLULE PEL MEDIANTE siRNA; OTTIMIZZAZIONE DEL SISTEMA DI TRANSFEZIONE E STUDI IN VITRO [Abstract in Atti di Convegno]
Belletti, Daniela; Riva, Giovanni; Tosi, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela; Ruozi, Barbara
abstract

Il trattamento del cancro mediante la chemioterapia tradizionale è spesso ostacolato dall’alta tossicità sistemica e dalla scarsa selettività dei principi attivi. La tecnologia siRNA (Small Interfering RNAs) basata sulla transfezione di ODN RNA antisenso, disegnati ad hoc per riconoscere e bloccare l’RNA messaggero target, rappresenta un’innovazione nelle strategie attuate in gene-silencing mostrando evidenti vantaggi anche in campo oncologico rispetto ai tradizionali chemioterapici. [1]Il linfoma effusivo delle cavità sierose (PEL), oggetto della ricerca, è un particolare tipo di linfoma associato invariabilmente all’infezione di HHV-8, con opzioni terapeutiche convenzionali limitate e non efficaci.[2] La disregolazione specifica mediata da siRNA del network di trascrizione della cellula malata rappresenta, ad oggi, una possibile alternativa nel trattamento della patologia. Considerando l’instabilità in vivo dei siRNAs associata alla loro scarsa capacità di penetrazione cellulare, il successo della strategia di silenziamento è strettamente dipendente dall’utilizzo di sistemi di delivery capaci di proteggere e trasportare selettivamente l’attivo nella cellula malata[3]. I liposomi, ed in particolare quelli cationici, sono sistemi di veicolazione e direzionamento innovativi, studiati e utilizzati da diversi anni, per la loro capacità di stabilizzare e trasferire materiale genico al bersaglio; la modificazione superficiale con molecole stabilizzanti (es PEG) e ligando selettive quali anticorpi monoclonali, peptidi ed aptameri garantisce la veicolazione e selettività del sistema [4,5].In studi preliminari, abbiamo dimostrato come sistemi liposomiali “stealth” modificati con anticorpo anti CD-138 specificamente riconosciuto da proteoglicani largamente espressi sulle cellule linfomatose, risultino altamente efficienti nella complessazione e stabilizzazione di materiale genico. Tali immunoliposomi sono stati testati quali carriers di un oligonucleotide modello (ODN-FITC) su cellule BCBL-1 (linea cellulare di PEL) evidenziando una buona capacità di trasferimento e di targeting valutata mediante citofluorimetria e microscopia confocale [6]. Tali evidenze sono risultate basilari per intraprendere nuovi studi di ottimizzazione di sistemi veicolanti siRNA specifici nel silenziamento di fattori trascrizionali dominanti dello stadio plasmacellulare (knock-down di BLIMP1/PRDM1) in cellule PEL. In questa ricerca sono stati formulati e caratterizzati vettori liposomiali (Lp) utilizzando lipidi neutri e cationici (Dotap e DC-Chol). Tali liposomi sono stati complessati con siRNAs e caratterizzati (elettroforesi su gel di agarosio, PCS ed AFM); i complessi ottenuti utilizzando il lipide cationico Dotap risultano caratterizzati da strutture definite e riproducibili, di dimensioni prossime a 350nm che efficientemente proteggono i siRNAs, mostrando inoltre una buona stabilità dopo incubazione in siero. I vettori sono stati testati in vitro su cellule BCBL-1; i dati preliminari evidenziano come i lipoplessi siano abili nel trasferire il materiale genico a livello citoplasmatico. L’inibizione della proteina bersaglio produce un effetto dose dipendente a concentrazioni superiori a 50nM evidente dopo 48/72 ore dal trattamento. In particolare è stata osservata un elevata induzione di necrosi /apoptosi mediante test annessina/propidio associata ad una significativa inibizione della proliferazione cellulare. E’ stata inoltre valutata la peghilazione come strategia per aumentare il tempo di permanenza in circolo oltre ad offrire il supporto per il legame con anticorpi specifici verso il PEL (anti.CD-138).[1] Oh YK, Park TG (2009). Adv. Drug Deliv Rev. 61: 850–862.[2] Carbone A, Gloghini A (2007). BJH review. 140: 13–24.[3] Whitehea KA , Langer R, Anderson DG (2009). Nature Review. 8:129-138.[4] Ruozi B, Belletti D, Tombesi A, Tosi G, Bondioli L, Forni F, Vandelli M A (2011). Int J Nanomed. 6:557–563.[5] Ruo

2011 - Massive bleeding: Are we doing our best? [Articolo su rivista]
Marietta, M; Pedrazzi, P; Girardis, Massimo; Luppi, Mario
abstract

Massive bleeding accounts for more than 50% of all trauma-related deaths within the first 48. h following hospital admission and it can significantly raise the mortality rate of any kind of surgery. Despite this great clinical relevance, evidence on the management of massive bleeding is surprisingly scarce, and its treatment is often based on empirical grounds. Successful treatment of massive haemorrhage depends on better understanding of the associated physiological changes as well as on good team work among the different specialists involved in the management of such a complex condition.

2011 - May the indirect effects of CIHHV-6 in transplant patients be exerted through the reactivation of the viral replicative machinery? [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Rossi, G; Riva, Giovanni; Vallerini, Daniela; Zanetti, Eleonora; Quadrelli, Chiara; Morselli, M; Forghieri, Fabio; Maccaferri, Monica; Paolini, Ambra; Marasca, Roberto; Narni, Franco; Luppi, Mario
abstract

Indirect effects of CIHHV-6 in transplant patients

2011 - Mucorles-specific T cells emerge in the course of invasive mucormucosis and may be used as a surrogate diagnostic marker in high-risk patients [Articolo su rivista]
Potenza, Leonardo; Vallerini, Daniela; Barozzi, Patrizia; Riva, Giovanni; Forghieri, Fabio; Zanetti, Eleonora; Quadrelli, Chiara; Candoni, A; Maertens, J; Rossi, Giulio; Morselli, M; Codeluppi, M; Paolini, Ambra; Maccaferri, Monica; DEL GIOVANE, Cinzia; D'Amico, Roberto; Rumpianesi, F; Pecorari, M; Cavalleri, F; Marasca, Roberto; Narni, Franco; Luppi, Mario
abstract

Mucorales-specific T cells have been investigated in 28 hematologic patients during the course of their treatment. Three developed proven invasive mucormycosis (IM), 17 infections of known etiologies but other than IM, and 8 never showed fever upon the period of observation. The Mucorales-specific T cells may be detected only in patients with IM, at diagnosis and along the entire course of the IM, but neither before nor long time after the resolution of the infection. Such T cells produced predominantly interleukin-4, interferon-gamma (IFN-γ), interleukin-10, and to a lesser extent also interleukin-17, and belonged to either CD4+ or CD8+ subsets. The specific T cells producing IFN-γ were able to directly induce damage of Mucorales hyphae. None of the 25 patients without IM showed Mucorales-specific T cells. Specific T cells contribute to human immune responses against fungi of the order Mucorales and could be evaluated as a surrogate diagnostic marker of IM.

2011 - Novel polymeric/lipidic hybrid systems (PLHs) for effective Cidofovir delivery: preparation, characterization and comparative in vitro study with polymeric particles and liposomes [Articolo su rivista]
Belletti, Daniela; Riva, Giovanni; Tosi, Giovanni; Forni, Flavio; Barozzi, Patrizia; Luppi, Mario; Vandelli, Maria Angela; Ruozi, Barbara
abstract

Cidofovir is an antiviral drug active as antitumoral agent a high doses against the Primary Effusion Lymphoma, a herpesvirus HHV8-associated B-cell lymphoma. A novel polymeric/lipidic hybrid system, consisting in a specific combination of biocompatible materials, capable to build a crossbred betweenpolymeric particles and liposomes were prepared and used to stabilize and deliver the drug, unsuccessfully formulated into several types of carriers. This innovative cidofovir-delivering system has structurally been characterized in comparison to multilamellar liposomes and polymeric particles, and then testedfor antitumoral efficacy against tumor cells (BCBL-1 cell line). The results demonstrated the improving of drug stability and encapsulation efficiency and suggested that polymeric/lipidic hybrid system could be promising to improve the antitumoral effect of cidofovir even at lower doses.

2011 - Three- or four-factor prothrombin complex concentrate for emergency anticoagulation reversal: what are we really looking for? [Articolo su rivista]
Marietta, M; Pedrazzi, P; Luppi, Mario
abstract

No abstract available

2011 - Tumor-targeted immunoliposomal nanosystems to deliver either Cidofovir or Antineoplastic SiRNA against Primary Effusion Lymphoma (PEL) [Relazione in Atti di Convegno]
Riva, Giovanni; Belletti, Daniela; Ruozi, Barbara; Barozzi, Patrizia; Vallerini, Daniela; Quadrelli, Chiara; Zanetti, Eleonora; Morselli, M; Forghieri, Fabio; Marasca, Roberto; Narni, Franco; Tosi, Giovanni; Forni, Flavio; Vandelli, Maria Angela; Potenza, Leonardo; Luppi, Mario
abstract

Therapeutic applications of siRNA-mediated gene silencing appear to be highly dependent on the use of pharmaco-technologic carrier systems, able to protect siRNAs from rapid degradation upon administration, as well as to specifically deliver them to target cells. Actually, while siRNA-expressing viral vectors are burdened with safety concerns for their clinical use, the development of modified liposomal nanocarriers may represent a feasible option to harness the therapeutic potential of targetedantineoplastic siRNAs. Recently, we have successfully developed and characterized effective immunoliposomal nanosystems (ILNs) for targeted delivery of Cidofovir (an anti-herpesviral nucleotideanalogue, also showing antitumor activity) against PEL cell lines, demonstrating a significant improvement of the antineoplastic activity of the drug, especially at lower doses (less than 1nM). Thus, we tried to adapt such PEL-specific ILNs (PEGilated nanovescicles made of cationic/neutral lipids, engineered with anti-CD138 moAb on their surface) to efficiently encapsulate siRNAs and deliver them into PEL cell lines (highly expressing CD138 membrane protein). Our preliminary data showed thatsingle treatments with anti-PEL ILNs, delivering specific siRNAs against Blimp1 (Prdm1), which is a master transcription factor in PEL (a plasmablast/pre-plasmacell lymphoma, consistently Bcl-6 neg, Blimp1 pos), were able to induce a dose-dependent (50-200nM) inhibition of Blimp1 production (as assessed by Blimp1 mRNA and protein levels using RT-PCR and Western Blot, respectively), and this was strongly associated with enhanced cell death (more than 80%, using Annexin V/PI test). In particular, we observed a massive reduction of PEL viability (mean viable cells 8%, range 3-15%) as soon as 48-72 hours after treatment with 100nM anti-Blimp1 siRNAs. Interestingly, these data may resemble those described in multiple myeloma cell lines, after transduction with lentiviral vector constitutively expressing anti-Blimp1 shRNAs. Further studies on PEL murine models are now warranted to assess the efficacy and toxicity profile of in-vivo treatment with PEL-specific ILNs, loadedwith either Cidofovir or anti-Blimp1 siRNAs.

2011 - β-HHVs and HHV-8 in Lymphoproliferative Disorders. [Articolo su rivista]
Quadrelli, Chiara; Barozzi, Patrizia; Riva, Giovanni; Vallerini, Daniela; E., Zanetti; Potenza, Leonardo; Forghieri, Fabio; Luppi, Mario
abstract

Similarly to Epstein-Barr virus (EBV), the human herpesvirus-8 (HHV-8) is a γ-herpesvirus, recently recognized to be associated with the occurrence of rare B cell lymphomas and atypical lymphoproliferations, especially in the human immunodeficiency virus (HIV) infected subjects. Moreover, the human herpesvirus-6 (HHV-6), a β-herpesvirus, has been shown to be implicated in some non-malignant lymph node proliferations, such as the Rosai Dorfman disease, and in a proportion of Hodgkin's lymphoma cases. HHV-6 has a wide cellular tropism and it might play a role in the pathogenesis of a wide variety of human diseases, but given its ubiquity, disease associations are difficult to prove and its role in hematological malignancies is still controversial. The involvement of another β-herpesvirus, the human cytomegalovirus (HCMV), has not yet been proven in human cancer, even though recent findings have suggested its potential role in the development of CD4(+) large granular lymphocyte (LGL) lymphocytosis. Here, we review the current knowledge on the pathogenetic role of HHV-8 and human β-herpesviruses in human lymphoproliferative disorders.

2010 - Adherence to international guidelines for the treatment of invasive aspergillosis in acute myeloid leukaemia: Feasibility and utility (SEIFEM-2008B study) [Articolo su rivista]
Pagano, Livio; Caira, Morena; Offidani, Massimo; Martino, Bruno; Candoni, Anna; Valentini, Caterina Giovanna; Specchia, Giorgina; Nosari, Annamaria; Tosti, Maria Elena; Leone, Giuseppe; Luppi, Mario; Aversa, Franco
abstract

Objectives and methods: In order to assess physicians' compliance with international guidelines for the targeted treatment of invasive aspergillosis, 136 patients with acute myeloid leukaemia and proven/probable invasive aspergillosis were analysed. Results: Compliance with Infectious Diseases Society of America (IDSA) and European Conference on Infections in Leukaemia (ECIL) guidelines was found to be relatively low (28% for ECIL and 55% for IDSA), although no significant differences were found between the two groups (adherence versus non-adherence). In both subgroup analyses (IDSA and ECIL), compliance with the guidelines did not impact the 120 day survival rate. Instead, adherence to guidelines led to a higher response rate to first-line antifungal treatment (76% in the IDSA group and 84% in the ECIL group). Conclusions: Guidelines establish categories of patients with homogeneous characteristics, and suggest optimal diagnostic and therapeutic options for them. Acquisition of good results through adherence to guidelines is confirmed by our series. Unfortunately, there are frequently reasons to deviate from these general recommendations, particularly in patients with acute myeloid leukaemia. Despite evidence-based recommendations, adherence to the guidelines does not constitute the best therapeutic choice in each and every patient. Subjects' clinical conditions and co-morbidities vary widely, and sometimes render the 'recommended' drug a non-applicable strategy. © The Author 2010. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.

2010 - Cidofovir-loaded liposomes: an intro-study using BCBL-1 cell line as a model for primary effusion lymphoma [Articolo su rivista]
Ruozi, Barbara; Riva, Giovanni; Belletti, Daniela; Tosi, Giovanni; Forni, Flavio; Mucci, Adele; Barozzi, Patrizia; Luppi, Mario; Vandelli, Maria Angela
abstract

Cidofovir (HPMPC) was recently reported to exert a valuable antineoplastic activity against primary effusion lymphoma (PEL), a B-cell neoplasm associated with Human Herpesvirus-8 (HHV-8) infection. In this study, we developed and characterized liposomes encapsulating HPMPC to increase drug efficacy reducing the administered dose and the related toxicity, which actually hamper its clinical therapeutic use in patients affected with PEL. The liposomes, obtained using different formulations of neutral and cationic lipids, were analyzed by microscopical (AFM) and spectroscopical (PCS and NMR) techniques. Using an in vitro model of PEL (BCBL-1 cell line), the carrier toxicity and the antineoplastic efficacy of liposomes were evaluated by flow cytometry applying apoptosis and cell death analysis. The in vitro study showed the applicability of the liposomes within a restricted range of lipidic concentrations according to the lipids used during the preparation. The moderate increases in the percentage of apoptotic/necrotic cells suggests that liposomal delivery allows the release of HPMPC into BCBL-1 cells enabling an unexpectedantineoplastic activity of this drug even at lower doses.

2010 - Common vascular endothelial growth factor variants and risk for posttransplant Kaposi sarcoma. [Articolo su rivista]
Zanetti, E; Barozzi, Patrizia; Brown, Ee; Bosco, R; Vallerini, Daniela; Riva, Giovanni; Quadrelli, Chiara; Potenza, Leonardo; Forghieri, Fabio; Montagnani, G; D'Amico, Roberto; Del Giovane, Cinzia; Duraes, C; Whitby, D; Machado, Jc; Schulz, Tf; Torelli, G; Luppi, Mario
abstract

No abstract available

2010 - Development and characterization of immunoliposomes for Cidofovir and SiRNA delivery: a new strategy for the treatment of Primary Effusion Lymphoma [Abstract in Atti di Convegno]
Belletti, Daniela; Ruozi, Barbara; Riva, Giovanni; Tosi, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela
abstract

Primary Effusion Lymphoma (PEL) is an aggressive and consistently lethal non-Hodgkin's B-cell lymphoma growing as lymphomatous effusions in serous body cavities and invariably associated with HHV-8 [1,2].The majority of the patients affected with PEL, either elderly HIV-negative or immunocompromised AIDS patients, are typically characterized by several age-related co-morbidities or opportunistic infectious diseases. Clinical efficacy of conventional anti-neoplastic chemotherapy is commonly hampered by the excessive grade of systemic toxicity and low drug levels in the tumor area [3].New therapeutic challenges may arise from the use of SiRNA technology, which is based on cellular transfection of antisense small RNAs, specifically designed to recognize the target mRNA and able to turn off the changed cellular mechanism of tumor cells, or by using antineoplastic drug (Cidofovir). Considering the high instability and the poor cellular uptake of both these actives, tumor-specific delivery by means of targeted nanocarriers is strongly required. These strategies represent an attractive approach to enhance intra-tumoral cytotoxic effects together with the reduction of “off-target” side effects, possibly offering a radical improvement in the treatment of such fragile oncologic patients. Among the colloidal carrier systems for drug delivery, liposomes have received considerable attention. They allow to protect the drug from rapid degradation, being particularly suitable to form complexes with highly-degradable ribonucleic acids.In this study, we formulated and characterized immunoliposomal formulation direct to PEL cell line (BCBL-1 cell line) using the cationic lipid DOTAP and a pegylated cholesterol funzionalized with a maleidoimide moiety capable to interact with anti CD-138 antibody. The formulation was characterized (size, zeta potential and morphology) in comparison with untargeted DOTAP liposomes and pegylated systems.These liposomal systems were used to transfect a model FITC-ODN into a model PEL cell line (BCBL-1). The studies on cellular binding and on the internalization of oligo by flow cytometry and confocal analysis confirmed the higher transfection efficiency of the immunoliposomes when compared with DOTAP and pegylated liposomes. This targeted formulation could be reasonably considered as optimal candidates for therapeutic siRNA delivery and more generally for gene encapsulation and delivery against the poor curable PEL tumor. Concerning cidofovir, it has been demonstrated that this antiviral drug is able to induce cell apoptosis in different tumor included PEL. Indeed, the high pro-apoptotic concentrations of cidofovir are never achievable in situ after full-dose systemic administration (5mg/Kg i.v.), and however, this systemic treatment can frequently cause severe nephrotoxicity [4]. We proposed liposomes encapsulating cidofovir by a modified reversed phase evaporation method (mREV) followed by extrusion. The characterization of samples suggested that cationic liposomes are more suitable for cidofovir stabilization, taking advantage of the charge interaction between the anionic drug and the cationic lipid moieties. Using the in vitro model of PEL (BCBL-1 cell line), the carrier toxicity and the antineoplastic efficacy of liposomes were evaluated by flow cytometry, applying apoptosis and cell death analysis. This in vitro study showed the applicability of the liposomes within a restricted range of lipidic concentrations, mainly depending on the lipids used during the preparation. The cidofovir transfection mediated by liposome composed of PC:DOTAP and PC:DC-CHOL caused a moderate increase in the percentage of apoptotic/necrotic PEL cells with respect to the controls (free drug and empty liposomes) suggesting that liposomal delivery allows the release of cidofovir into BCBL-1 cells enabling an unexpected antineoplastic activity of this drug even at lower doses.

2010 - Emergence of BCR-ABL-specific cytotoxic T cells in the bone marrow of patients with Ph+ acute lymphoblastic leukemia during long-term Imatinib mesylate treatment. [Articolo su rivista]
Riva, Giovanni; Luppi, Mario; Barozzi, Patrizia; Quadrelli, Chiara; Basso, S; Vallerini, Daniela; Zanetti, Eleonora; Morselli, M; Forghieri, Fabio; Maccaferri, M; Volzone, F; DEL GIOVANE, Cinzia; D'Amico, Roberto; Locatelli, F; Torelli, Giuseppe; Comoli, P; Potenza, Leonardo
abstract

Imatinib mesylate (IM) has been demonstrated to be permissive for the emergence of T cells directed against chronic myeloid leukemia cells. Ten Philadelphia chromosome-positive acute lymphoblastic leukemia patients, receiving high dose IM maintenance, underwent a long-term immunological monitoring (range 2-65 months) of (p190)BCR-ABL-specific T cells in the bone marrow (BM) and peripheral blood (PB). (p190)BCR-ABL-specific T lymphocytes were detected in all patients, more frequently in BM than in PB samples (67% vs 25%, p&lt;0.01), and resulted significantly associated with lower minimal residual disease values (p&lt;0.001), while absent at leukemia relapse. Specific T cells were mainly effector memory CD8+ and CD4+ T cells, producing IFNgamma, TNFalpha and IL-2 (median % positive cells: 3.34, 3.04, 3.58, respectively). Cytotoxic subsets able to lyse BCR-ABL-positive leukemia blasts were also detectable. Whether these autologous (p190)BCR-ABL-specific T cells may be detectable under other tyrosine-kinase inhibitors, expanded ex vivo and exploited for immunotherapy remains to be addressed.

2010 - Formulation and characterization of new Polymeric/Lipidic Hybrid systems for cidofovir delivery [Abstract in Atti di Convegno]
Belletti, Daniela; Riva, Giovanni; Tosi, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela; Ruozi, Barbara
abstract

Cidofovir is an antiviral drug with a remarkable antitumour activity in several animal models, associated or not with viral infections [1]. More recently, cidofovir has also been shown to be an effective treatment against Primary Effusion Lymphoma (PEL), a B cell non–Hodgkin lymphoma involving the serous cavities, invariably associated with Human HerpesVirus-8 (HHV8) and often with Epstein–Barr Virus (EBV) infection. A hindrance to the clinical applications of cidofovir is the high systemic toxicity, mainly the nephrotoxicity. Cidofovir encapsulation into specific micro- or nanocarriers, able to extend the release of the drug, may represent an effective strategy both to minimize the off-target organ exposure as well as to simultaneously increase the concentration of drug within the site of action. Unfortunately, the physical-chemical characteristics of the drug (low molecular weight, high solubility at different pH, unfavourable partition coefficient) limit the encapsulation into delivery systems [2,3]. Recently, we proposed the use of cationic liposomes to stably encapsulate cidofovir; unfortunately, the in vivo applicability of such cidofovir carriers is limited by the presence of cationic lipids, inducing a dose-related toxicity [4]. To overcome this problem, in this work we aimed to investigate a novel hybrid system, consisting in a specific combination of biocompatible materials, capable to build a crossbred between polymeric particles and liposomes. This innovative cidofovir-delivering systems (called PLHs, polymeric/lipidic hybrid systems) made of phosphatidilcholine (PC), cholesterol (CHOL) and polylactic acid (PLA) have been characterized (size, zeta potential, morphology, structure and thermal behaviour) in comparison to multilamellar liposomes and polymeric particles. Microscopical studies (atomic force and confocal microscopy), in agreement with the other characterizations, suggested that a rearrangement of the components has taken place to form a new matricial porous structure different both from liposomes and polymeric particles, with a wide dispersion of polymer in the lipidic bulk. This new crossbeald delivery systems was able to increased the encapsulation of cidofovir (encapsulation efficiency twice higher than liposomes and about 10 times higher than polymeric particles) and resulted atossic against PEL tumor cells (BCBL-1 cell line) revealing also a capability to better traslocate the drug into the cells causing and increased apoptosis respect to the free drug.

2010 - HHV-6 and atypical lymphoproliferative disorders: are only qualitative molecular examinations sufficient to support a pathogenetic role? [Articolo su rivista]
Forghieri, Fabio; Potenza, Leonardo; Barozzi, Patrizia; Vallerini, Daniela; Riva, Giovanni; Zanetti, Eleonora; Quadrelli, Chiara; Torelli, Giuseppe; Luppi, Mario
abstract

n.d.

2010 - High versus standard dose methylprednisolone in the acute phase of idiopathic thrombotic thrombocytopenic purpura: a randomized study. [Articolo su rivista]
Balduini, Cl; Gugliotta, L; Luppi, Mario; Laurenti, L; Klersy, C; Pieresca, C; Quintini, G; Iuliano, F; Re, R; Spedini, P; Vianelli, N; Zaccaria, A; Pogliani, Em; Musso, R; Bobbio Pallavicini, E; Quarta, G; Galieni, P; Fragasso, A; Casella, G; Noris, P; Ascari, E; The, Italian TTP Study Group
abstract

Therapeutic plasma exchange (PE) is the accepted therapy for thrombotic thrombocytopenic purpura (TTP). Because not all patients achieve remission, other treatment modalities have been used in addition to PE, but no randomized clinical trial evaluated their efficacy. The aim of this multicentric study was to compare the effectiveness of standard- versus high-dose methylprednisolone as an adjunctive treatment to PE in the acute phase of TTP. Sixty patients with idiopathic TTP were randomized to receive methylprednisolone 1 mg/kg/die intravenous or 10 mg/kg/die for 3 days, thereafter, 2.5 mg/kg/die in addition to PE. Both dosages of steroids were well tolerated. At the end of induction therapy (day 23), the percentage of patients failing to achieve complete remission was significantly higher in the standard dose (16 of 30) than in the high-dose group (seven of 30). Also, the number of cases without a good response at day 9 and the number of deaths were higher in the standard-dose arm, but the differences did not reach the statistical significance. Results of present study indicate that the association of PE with high-dose instead of standard-dose steroids reduces the percentage of TTP patients that fail to achieve complete remission.

2010 - Immunoliposomal systems targeting the primary effusion lymphoma (PEL): in vitro study [Articolo su rivista]
Ruozi, Barbara; Riva, Giovanni; Belletti, Daniela; Tosi, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela
abstract

Aims: To develop an appropriate liposomal formulation for gene delivery against Primary Effusion Lymphoma (PEL), a herpesvirus HHV8-associated B-cell lymphoma. Materials and methods: Cationic, cationic pegylated and cationic pegylated anti-CD138 liposomes (ILp) linking a monoclonal antibody expressed on PEL cells were prepared by thin layer evaporation method followed by extrusion technique. The formulations were mixed with a model oligonucleotide to form the lipoplexes tested on BCBL-1 cell (a PEL cell line). The transfection efficiency was evaluated by flow cytometry and confocal laser scanning microscopy analysis. Results: Based on antigen–antibody interaction, ILp mediated a specific gene delivery as shown by a significant increase in the transfection rate and a localized internalization of the oligo, in comparison with cationic liposomes and cationic pegylated liposomes.Conclusion: ILp could be proposed as effective carriers for oligo transfer in BCBL-1 cells. In vitro experimental results encourage to further test the in vivo therapeutic potentials of ILp for specific delivery of antitumoral agents.

2010 - Immunoliposomes for the delivery of SiRNA and chemioterapeutic agent to primary effusion lymphoma [Abstract in Atti di Convegno]
Belletti, Daniela; Ruozi, Barbara; Riva, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela
abstract

In this study we have formulated and characterized immunoliposomes direct to PEL cells using the cationic lipid DOTAP and a pegylated cholesterol funzionalized with a maleidoimide moiety capable to stable interact with anti CD 138 antbody. The formulation was characterized in comparison with only pegylated systems and DOTAP liposomes. These liposomes were used to transfect a model of FITC-ODN into the target cells. The efficiency of these carriers was evaluated using flow cytometry and confocal analysis. The results confirm the higher transfection efficiency of the immunoliposomes if compared with conventional and pegylated liposomes.

2010 - Immunological and inflammatory features of Kaposi's sarcoma and other Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8-associated neoplasias. [Articolo su rivista]
Riva, Giovanni; Barozzi, Patrizia; Torelli, Giuseppe; Luppi, Mario
abstract

During the last 15 years, virologic and immunologic studies have provided a series of valuable clues on the modalities of gamma-herpesvirus-induced oncogenesis, which do not only consist of the direct subversion of intracellular signaling pathways, leading to a frank neoplastic molecular network in the infected cell, but also rely on viral manipulations of the cellular and cytokine microenvironment, especially in conditions of immunodeficiency in the host. At the virus-host interface, something iniquitous, strikingly favoring the aggressive expansion of human herpesvirus 8-infected lympho-endothelial clones, known as Kaposi's sarcoma, often occurs in different types of immunocompromised patients, able to establish a deleterious "pro-Kaposi's sarcoma" neo-angiogenic inflammatory network. However, these patients may control - or even resolve - the neoplastic burden as soon as an immunologic reassessment restores functional anti-Kaposi's sarcoma immune responses and reconstitutes a proper inflammatory environment. Indeed, the occurrence of iatrogenic Kaposi's sarcoma remissions, after the reduction or switch of immunosuppressive regimens, strongly suggests that the reset of immunologic constraints characterizing the Kaposi's sarcoma onco-pathogenic system may be sufficient to inhibit human herpesvirus 8-positive lympho-endothelial proliferations. Accordingly, immunologic reports all underline the pivotal protective role of anti-human herpesvirus 8 memory T-cells (harmonically, both CD8+ and CD4+ subsets), thus definitely implying a general requirement for an effective, antiviral immuno-inflammatory environment, based on correct and productive interactions between different compartments of dendritic, myeloid, and specific T-cells, in order to achieve and maintain optimal control on human herpesvirus 8-associated antigenic stimulations and Kaposi's sarcoma disease. In this review, we recapitulate some remarkable features about the outstanding immunologic issue raised by human herpesvirus 8-driven neoplastic outgrowths in immunodeficient patients, and in particular, we discuss the emerging view of Kaposi's sarcoma as an atypical neoplastic process, tightly dependent on immune system dynamics. It is conceivable that functional dissection of the specific immune responses, capable to cope with human herpesvirus 8, and further definitions of a global inflammatory profile with protective activity against Kaposi's sarcoma outbreaks, will eventually foster immunologic monitoring protocols during the follow-up of AIDS and posttransplant patients, either preventing or treating human herpesvirus 8-related tumors by multifunctional immunomodulation or prompt development of adoptive immunotherapeutic approaches.

2010 - Impact of setting of care on pain management in patients with cancer: a multicentre cross-sectional study. [Articolo su rivista]
Sichetti, D; Bandieri, E; Romero, M; Di Biagio, K; Luppi, Mario; Belfiglio, M; Tognoni, G; Ripamonti, Ci; ECAD Working, Group
abstract

BACKGROUND: No study has so far addressed whether differences do exist in the management of cancer-related pain in patients admitted to oncology and non-oncology settings.PATIENTS AND METHODS: A multicentre cross-sectional study in 48 Italian hospitals has enrolled 819 patients receiving analgesic therapy for cancer-related pain. Demographics and clinical and analgesic therapy information have been prospectively collected by standardized forms. Adequacy of pain management has been evaluated by the Pain Management Index (PMI).RESULTS: Differences in the analgesic drug administration according to settings of care have been evident, non-opioids more frequently being administered in non-oncology units (19.6% versus 7.0%; P < 0.0001), while strong opioids are more frequently used in the oncology units (69.5% versus 51.9%; P < 0.0001). The number of patients receiving inadequate therapy (PMI < 0) has lowered in oncology compared with non-oncology units (11.3% versus 18.8%; P = 0.0024). Results of multiple logistic regression analysis have shown that the admission to non-oncology setting [odds ratio (OR) = 1.75, 95% confidence interval (CI) = 1.15-2.67; P = 0.0096] and the absence of metastatic disease (OR = 1.60, 95% CI = 1.04-2.44; P = 0.0317) were independent factors associated with an increased risk of receiving an inadequate analgesic therapy.CONCLUSION: Oncology wards provide the most adequate standard of analgesic therapy for cancer-related pain.

2010 - Is pain in patients with haematological malignancies under-recognised? The results from Italian ECAD-O survey. [Articolo su rivista]
Bandieri, E; Sichetti, D; Luppi, Mario; Di Biagio, K; Ripamonti, C; Tognoni, G; Belfiglio, M; Romero, M.
abstract

No abstract available

2010 - Nanosistemi lipidici targettizzati per il direzionamento di siRNA con attività antineoplastica al linfoma primitivo delle cavità sierose (PEL) [Abstract in Atti di Convegno]
Belletti, Daniela; Ruozi, Barbara; Riva, Giovanni; Tosi, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela
abstract

La tecnologia dei siRNA (Small Interfering RNAs) basata sulla transfezione di ODN RNA antisenso, disegnata ad hoc per riconoscere e bloccare l’RNA messaggero target, rappresenta l’innovazione nelle strategie attuate in gene-silencing con importanti prospettive per il trattamento di patologie complesse, come il linfoma primitivo delle cavità sierose (PEL), oggetto della ricerca.Il PEL è un particolare tipo di linfoma associato invariabilmente all’infezione di HHV-8, altamente aggressivo che presenta opzioni terapeutiche convenzionali limitate e non efficaci. La disregolazione specifica mediata da siRNA del network di trascrizione della cellula malata rappresenta, ad oggi, una possibile alternativa nel trattamento della patologia. I liposomi, ed in particolare quelli cationici, sono da diversi anni studiati per la loro capacità di stabilizzare materiale genico proteggendolo dalla degradazione in vivo ed offrendo inoltre la possibilità di coniugare molecole specifiche nel riconoscimento del target.In questa ricerca sono stati formulati e caratterizzati sistemi immunoliposomiali “stealth” e targettizzati con anticorpo CD-138 specificamente riconosciuto da proteoglicani largamente espressi sulle cellule linfomatose. Tali immunoliposomi sono stati testati in studi preliminari quali carriers di un oligonucleotide modello in una linea cellulare di PEL (BCBL-1); la capacità di trasferimento e di targeting è stata valutata mediante citofluorimetria e microscopia confocale. Successivamente sono stati allestiti immunoliposomi incorporanti siRNA per il silenziamento di fattori trascrizionali dominanti dello stadio plasmacellulare (knock-down di BLIMP1/PRDM1) e testati sulla stessa linea cellulare. I risultati evidenziano come i nostri sistemi siano altamente efficienti nella stabilizzazione del materiale genico promuovendo inoltre il trasferimento a livello citoplasmatico dove avviene il silenziamento della proteina bersaglio con conseguente apoptosi cellulare.

2010 - Naturally occurring human urinary peptides for use in diagnosis of chronic kidney disease. [Articolo su rivista]
Good, Dm; Zürbig, P; Argilés, A; Bauer, Hw; Behrens, G; Coon, Jj; Dakna, M; Decramer, S; Delles, C; Dominiczak, Af; Ehrich, Jh; Eitner, F; Fliser, D; Frommberger, M; Ganser, A; Girolami, Ma; Golovko, I; Gwinner, W; Haubitz, M; Herget Rosenthal, S; Jankowski, J; Jahn, H; Jerums, G; Julian, Ba; Kellmann, M; Kliem, V; Kolch, W; Krolewski, As; Luppi, Mario; Massy, Z; Melter, M; Neusüss, C; Novak, J; Peter, K; Rossing, K; Rupprecht, H; Schanstra, Jp; Schiffer, E; Stolzenburg, Ju; Tarnow, L; Theodorescu, D; Thongboonkerd, V; Vanholder, R; Weissinger, Em; Mischak, H; Schmitt Kopplin, P.
abstract

Because of its availability, ease of collection, and correlation with physiology and pathology, urine is an attractive source for clinical proteomics/peptidomics. However, the lack of comparable data sets from large cohorts has greatly hindered the development of clinical proteomics. Here, we report the establishment of a reproducible, high resolution method for peptidome analysis of naturally occurring human urinary peptides and proteins, ranging from 800 to 17,000 Da, using samples from 3,600 individuals analyzed by capillary electrophoresis coupled to MS. All processed data were deposited in an Structured Query Language (SQL) database. This database currently contains 5,010 relevant unique urinary peptides that serve as a pool of potential classifiers for diagnosis and monitoring of various diseases. As an example, by using this source of information, we were able to define urinary peptide biomarkers for chronic kidney diseases, allowing diagnosis of these diseases with high accuracy. Application of the chronic kidney disease-specific biomarker set to an independent test cohort in the subsequent replication phase resulted in 85.5% sensitivity and 100% specificity. These results indicate the potential usefulness of capillary electrophoresis coupled to MS for clinical applications in the analysis of naturally occurring urinary peptides.

2010 - Organising pneumonia mimicking invasive fungal disease in patients with leukaemia. [Articolo su rivista]
Forghieri, Fabio; Potenza, Leonardo; Morselli, M; Maccaferri, Monica; Pedrazzi, L; Barozzi, Patrizia; Vallerini, Daniela; Riva, Giovanni; Zanetti, Eleonora; Quadrelli, Chiara; Rossi, G; Rivasi, Francesco; Messino', M; Rumpianesi, F; Grottola, Antonella; Venturelli, C; Pecorari, M; Codeluppi, M; Torelli, Giuseppe; Luppi, Mario
abstract

Clinical charts from 63 consecutive highly immunocompromised haematologic patients presenting with pulmonary nodular lesions on CT scan, classified as either probable or possible invasive fungal disease (IFD) according to the revised EORTC/MSG classification, were retrospectively studied. Histopathological analysis of lung tissues, available for 23 patients, demonstrated proven IFD in 17 cases (14 invasive aspergillosis and 3 invasive zygomycosis), diffuse alveolar damage in one and organising pneumonia (OP) in five cases. In the OP cases, three of which have been defined as probable IFD according to EORTC/MSG classification, extensive immunohistochemical, molecular and immunological analyses for fungi were negative. Our case descriptions extend the notion that OP may be encountered as a distinct histopathological entity in pulmonary nodular lesions in patients with leukaemia with probable/possible IFD.

2010 - Pain and emotional distress in leukemia patients at diagnosis. [Articolo su rivista]
Morselli, M; Bandieri, E; Zanin, R; Buonaccorso, L; D'Amico, Roberto; Forghieri, Fabio; Pietramaggiori, A; Potenza, Leonardo; Berti, A; Cacciapaglia, G; Molitierno, A; Galli, L; Artioli, F; Ripamonti, C; Bruera, E; Torelli, Giuseppe; Luppi, Mario
abstract

Pain and emotional distress in leukemia patients at diagnosis.

2010 - Review Part 3: Human herpesvirus-6 in multiple non-neurological diseases. [Articolo su rivista]
Ablashi, Dv; Devin, Cl; Yoshikawa, T; Lautenschlager, I; Luppi, Mario; Kühl, U; Komaroff, Al
abstract

No abstract available

2010 - Splenic hyalohyphomycosis, molecularly and immunologically consistent with invasive aspergillosis, in a patient with acute lymphoblastic leukemia. [Articolo su rivista]
Forghieri, Fabio; Rossi, G; Potenza, Leonardo; Morselli, M; Barozzi, Patrizia; Vallerini, Daniela; Messino, M; Rumpianesi, F; Pecorari, M; Torelli, Giuseppe; Luppi, Mario
abstract

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2010 - The therapeutic response and clinical outcome of adults with ALL1(MLL)/AF4 fusion positive acute lymphoblastic leukemia according to the GIMEMA experience. [Articolo su rivista]
Cimino, G; Cenfra, N; Elia, L; Sica, S; Luppi, Mario; Meloni, G; Vignetti, M; Paoloni, F; Foà, R; Mandelli, F.
abstract

The clinical outcome of 21 adults with ALL1(MLL)/AF4 positive acute lymphoblastic leukemia enrolled in the GIMEMA LAL 2000 trial and of 25 patients entered into the previous 0496 study is reported. LAL 2000 included more intensive consolidation and transplants. Complete remission rates were 90% and 88% in the LAL 2000 and 0496 trials, respectively. Fifteen patients were transplanted (5 autologous, 10 allogeneic). At 36 months overall and disease free survivals were 32.9%, 31.8%, 28% and 27.3%, in LAL 2000 and 0496 trials, respectively. Relapses remained the main reason of failure occurring in 10 and 16 of the 19 and 22 responding patients. In the LAL 2000 study, 4 relapses were observed before transplant. Thus, ALL1(MLL)/AF4 abnormality characterized a subset of patients with adverse prognosis in which the overall strategy adopted in the LAL 2000 study, rather than transplants "per se", failed to improve the patient clinical outcome.

2010 - Translational challenges of human herpesvirus 6 chromosomal integration. [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Torelli, G; Luppi, Mario
abstract

No abstract available

2010 - invasive aspergillosis in patients with acute myeloid leukemia: SEIFEM-2008 registry study. [Articolo su rivista]
Pagano, L; Caira, M; Candoni, A; Offidani, M; Martino, B; Specchia, G; Pastore, D; Stanzani, M; Cattaneo, C; Fanci, R; Caramatti, C; Rossini, F; Luppi, Mario; Potenza, Leonardo; Ferrara, F; Mitra, Me; Fadda, Rm; Invernizzi, R; Aloisi, T; Picardi, M; Bonini, A; Vacca, A; Chierichini, A; Melillo, L; de Waure, C; Fianchi, L; Riva, M; Leone, G; Aversa, F; Nosari, A.
abstract

Background This study evaluated prognostic factors, treatments and outcome of invasive aspergillosis (IA) in patients with acute myeloid leukemia (AML). DESIGN AND METHODS: The registry, which was activated in 2004 and closed in 2007, collected data on AML patients admitted to 21 hematological divisions in tertiary care centres or university hospitals in Italy, who developed proven or probable IA. RESULTS: 140 cases of IA were collected, with most cases occurring in post-induction aplasia, the highest risk phase in AML. IA-attributable mortality rate was 27%, confirming previous reports of a downward trend. Univariate and multivariate analyses revealed AML stage, duration of, and recovery from, neutropenia as independent prognostic factors. We analyzed outcomes after treatment with the three most frequently used drugs (liposomal amphotericin B, caspofungin, voriconazole). No differences emerged in survival on the 120th day or in the overall response rate which was 71%, ranging from 61% with caspofungin to 84% with voriconazole. Conclusions Our series confirms the downward trend in mortality rates reported in previous series, with all new drugs providing similar survival and response rates. Recovery from neutropenia and disease stage are crucial prognostic factors. Efficacious antifungal drugs bridge the gap due to poor hematological and immunological reconstitution.

2009 - Acute promyelocytic leukemia in very elderly patients: still aclinical challenge. [Articolo su rivista]
Forghieri, Fabio; Luppi, Mario; Morselli, M; Favale, V; Potenza, Leonardo; Volzone, F; Maccaferrim, ; Torelli, Giuseppe
abstract

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2009 - Changes in T-Cell Responses Against Human Herpesvirus-8 Correlate with the Disease Course of Iatrogenic Kaposi's Sarcoma in a Patient with Undifferentiated Arthritis [Articolo su rivista]
Barozzi, Patrizia; Potenza, Leonardo; Riva, Giovanni; Vallerini, Daniela; Quadrelli, Chiara; Bosco, Raffaella; Morselli, M; Forghieri, F; Volzone, F; Rossi, G; Ferri, Clodoveo; Bovini, C; Ciceri, F; Bordignon, C; Whitby, D; Schulz, Tf; Torelli, Giuseppe; Luppi, Mario
abstract

OBJECTIVES: To describe the first in-depth analysis of both the T-cell responses against human herpesvirus-8 (HHV-8) and the HHV-8 viral load in 1 patient who developed iatrogenic HHV-8-associated-Kaposi's sarcoma (KS) following immunosuppressive treatment for undifferentiated arthritis and to review the literature on iatrogenic KS (IKS). METHODS: T-cell responses against HHV-8 lytic and latent antigens were analyzed by ex vivo enzyme-linked immunospot (Elispot) and HHV-8 viral load was assessed by quantitative polymerase chain reaction, in sequential peripheral blood samples from a 55-year-old woman who developed skin/mucosal and visceral KS, while receiving treatment with cyclosporine, methotrexate, and methylprednisolone for undifferentiated arthritis. RESULTS: KS may result from HHV-8 infection in patients undergoing immunosuppressive treatment for rheumatic diseases and this is the first case of IKS occurring in undifferentiated arthritis. A role for immune surveillance in the pathogenesis of IKS is supported by the observation of disease regression following discontinuation of immunosuppressive therapy. In a 4-year follow-up, we showed that variations of the virus-specific immune responses but not of the viral load correlated well with the disease course, characterized by 2 remission and subsequent relapse phases, following changes of immunosuppressive therapy. CONCLUSIONS: We have provided evidence of a clear-cut correlation between changes in immunologic markers of HHV-8 infection and the disease course of this viral associated tumor, concomitant with variations of immunosuppressive treatment. Thus, ex vivo enzyme-linked immunospot for HHV-8-specific T-cell responses represents a new tool for the clinical management of rheumatic patients with IKS.

2009 - Diagnosis of aspergillosis: Role of proteomics [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Vallerini, Daniela; Zanetti, Eleonora; Torelli, Giuseppe; Luppi, Mario
abstract

The expansion of the antifungal armamentarium and the implementation of imaging techniques and new nonculture-based fungal diagnostics (NCBFDs) have improved the survival of patients with invasive aspergillosis (IA). However, mortality rates still remain high, possibly influenced by several pitfalls, affecting NCBFDs and reducing the window of opportunity for earlier treatment. A large body of in vitro and in vivo studies has demonstrated that several fungal proteic components are strongly immunogenic, and both the adaptive immunity and the innate branch are heavily involved in the recognition and clearance of fungal pathogens, resulting, on occasion, in a useful tool for the treatment of IA. By evaluating these studies, this review considers the possibility of exploiting either components of the innate or adaptive immunity to support the rapid and early diagnosis of IA. Copyright © 2009 by Current Medicine Group LLC.

2009 - Immunoliposomes for the delivery of SiRNA and chemioterapeutic agent to primary effusion lymphoma [Abstract in Atti di Convegno]
Belletti, Daniela; Ruozi, Barbara; Riva, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela
abstract

Breve descrizione delle metodologie attuate per promuovere il rilascio di farmaci e materiale genico al tumore. In particolare è stato discusso l'approccio con liposomi e sistemi lipidici

2009 - Immunoliposomes to target the Primary Effusion Lymphoma (PEL): in vitro study [Abstract in Atti di Convegno]
Belletti, Daniela; Ruozi, Barbara; Riva, Giovanni; Barozzi, Patrizia; Luppi, Mario; Forni, Flavio; Vandelli, Maria Angela
abstract

Preparazione e caratterizzazione di immunoliposomi per la veicolazione di materiale genico ai tumori (limfoma effusivo primario)

2009 - Indirect Antitumor Effects of Mammalian Target of Rapamycin Inhibitors Against Kaposi Sarcoma in Transplant Patients [Articolo su rivista]
Barozzi, Patrizia; Riva, Giovanni; Vallerini, Daniela; Bosco, Raffaella; Quadrelli, Chiara; Zanetti, Eleonora; Potenza, Leonardo; Forghieri, Fabio; Torelli, Giuseppe; Luppi, Mario
abstract

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2009 - Inflammatory myofibroblastic tumor of the bone marrow. [Articolo su rivista]
Saviola, Alessia; Rossi, Giulio; Bonacorsi, G; Forghieri, Fabio; Fiorani, C; Artusi, Tullio; Emilia, Giovanni; Luppi, Mario; Longo, G; Torelli, Giuseppe
abstract

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2009 - Interferon-alpha may restore sensitivity to tyrosine-kinase inhibitors in Philadelphia chromosome positive acute lymphoblastic leukaemia with F317L mutation [Articolo su rivista]
Potenza, Leonardo; Volzone, F; Riva, Giovanni; Soverini, S; Martinelli, S; Iacobucci, I; Gnani, A; Barozzi, Patrizia; Forghieri, Fabio; Morselli, M; Zanetti, E; Maccaferri, Monica; Baccarani, M; Martinelli, G; Torelli, Giuseppe; Luppi, Mario
abstract

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2009 - Performance of tests for latent tuberculosis in different groups of immunocompromised patients. [Articolo su rivista]
Richeldi, Luca; Losi, M; D'Amico, Roberto; Luppi, M; Ferrari, A; Mussini, Cristina; Codeluppi, M; Cocchi, S; Prati, F; Paci, V; Meacci, M; Meccugni, B; Rumpianesi, F; Roversi, P; Cerri, Stefania; Luppi, F; Ferrara, G; Latorre, I; Gerunda, Giorgio Enrico; Torelli, G; Esposito, R; Fabbri, Leonardo
abstract

BACKGROUND: Immunocompromised persons infected with Mycobacterium tuberculosis (MTB) have increased risk of tuberculosis (TB) reactivation, but their managementis hampered by the occurrence of false-negative results of the tuberculin skin test (TST). The T-cell interferon (IFN)-gamma release blood assays T-SPOT.TB(TS.TB) [Oxford Immunotec; Abingdon, UK] and QuantiFERON-TB Gold In-Tube (QFT-IT) [Cellestis Ltd; Carnegie, VIC, Australia] might improve diagnostic accuracy forlatent TB infection (LTBI) in high-risk persons, although their performance in different groups of immunocompromised patients is largely unknown.METHODS AND RESULTS: Over a 1-year period, we prospectively enrolled patients in three different immunosuppressed groups, as follows: 120 liver transplantation candidates (LTCs); 116 chronically HIV-infected persons; and 95 patients with hematologic malignancies (HMs). TST, TS.TB, and QFT-IT were simultaneouslyperformed, their results were compared, and intertest agreement was evaluated.Overall, TST provided fewer positive results (10.9%) than TS.TB (18.4%; p <0.001) and QFT-IT (15.1%; p = 0.033). Significantly fewer HIV-infected individuals had at least one positive test (9.5%) compared with LTCs (35.8%; p < 0.001) and patients with HMs (29.5%; p < 0.001). Diagnostic agreement between tests was moderate (kappa = 0.40 to 0.65) and decreased in the HIV-infected group when the results of the TS.TB were compared with either TST (kappa = 0.16) orQFT-IT (kappa = 0.19). Indeterminate blood test results due to low positive control values were significantly more frequent with QFT-IT (7.2%) than with TS.TB (0.6%; p < 0.001).CONCLUSIONS: Blood tests identified significantly more patients as being infected with MTB than TST, although diagnostic agreement varied across groups. Based onthese results, we recommend tailoring application of the new blood IFN-gamma assays for LTBI in different high-risk groups and advise caution in their current use in immunosuppressed patients.

2009 - Persistent hiccups as an adverse event to FLAG-IDA regimenfor leukemia. [Articolo su rivista]
Forghieri, Fabio; Maccaferri, M; Morselli, M; Potenza, Leonardo; Volzone, F; Bandieri, E; Torelli, Giuseppe; Luppi, Mario
abstract

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2009 - Prescription of opioids in Italy: everything, but the morphine. [Articolo su rivista]
Bandieri, E; Chiarolanza, A; Luppi, Mario; Magrini, N; Marata, Am; Ripamonti, C.
abstract

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2009 - Prevalence of Human Herpesvirus-6 Chromosomal Integration (CIHHV-6) in Italian Solid Organ and Allogeneic Stem Cell Transplant Patients [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Masetti, M; Pecorari, M; Bresciani, P; Gautheret Dejean, A; Riva, Giovanni; Vallerini, Daniela; Tagliazucchi, S; Codeluppi, M; DI BENEDETTO, Fabrizio; Gerunda, Giorgio Enrico; Narni, Franco; Torelli, Giuseppe; Luppi, Mario
abstract

The unique phenomenon of human herpesvirus-6 (HHV-6) chromosomal integration (CIHHV-6) may account for clinical drawbacks in transplant setting, being misinterpreted as active infection and leading to unnecessary and potentially harmful treatments. We have investigated the prevalence of CIHHV-6 in 205 consecutive solid organ (SO) and allogeneic stem cell transplant (alloSCT) Italian patients. Fifty-two (38.5%) of 135 solid organ transplant (SOT) and 16 (22.8%) of 70 alloSCT patients resulted positive for plasma HHV-6 DNA by real-time polymerase chain reaction. Seven SOT and three alloSCT patients presented HHV-6-related diseases, requiring antivirals. Two further patients (0.9%) were identified, presenting high HHV-6 loads. The quantification of HHV-6 on hair follicles disclosed the integrated state, allowing the discontinuation of antivirals. Before starting specific treatments, CIHHV-6 should be excluded in transplant patients with HHV-6 viremia by the comparison of HHV-6 loads on different fluids and tissues. Pretransplantation screening of donors and recipients may further prevent the misdiagnosis of CIHHV-6.

2008 - A possible role for low-dose cyclosporine in refractory immune thrombocytopenic purpura. [Articolo su rivista]
Emilia, Giovanni; Luppi, Mario; Morselli, M; Forghieri, F; Potenza, Leonardo; Torelli, Giuseppe
abstract

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2008 - Acute appendicitis in adult neutropenic patients with hematologic malignancies [Articolo su rivista]
Forghieri, Fabio; Luppi, Mario; Narni, Franco; Morselli, M.; Potenza, Leonardo; Bresciani, Paola; Volzone, Francesco; Rossi, Giulio; Rossi, Aldo; Trenti, Loris; Barozzi, Patrizia; Torelli, Giuseppe
abstract

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2008 - Assessment of aspergillus-specific T cells for diagnosis of invasive aspergillosis in a leukemic child with liver lesions mimicking hepatosplenic candidiasis [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Rossi, Giulio; Palazzi, G; Vallerini, Daniela; Riva, Giovanni; Cellini, M; Morselli, M; Volzone, Francesco; Venturelli, C; Quadrelli, Chiara; Di Pancrazio, L; Cano, Mc; Paolucci, Paolo; Torelli, Giuseppe; Luppi, Mario
abstract

A child with acute myeloid leukemia presented with multiple liver lesions mimicking hepatosplenic candidiasis during the neutropenic phase following the induction chemotherapy. All the available diagnostic tools showed repeatedly negative results, including galactomannan. An enzyme-linked immunospot (ELISPOT) assay showed a high number of Aspergillus-specific T cells producing interleukin-10 [TH2(IL-10)] and a low number of Aspergillus-specific T cells producing gamma interferon [TH1(IFN-γ)], revealing invasive aspergillosis (IA) before the confirmatory biopsy. A progressive skewing from the predominance of TH2(IL-10) to a predominance of TH1(IFN-γ) was observed close to the complete resolution of the infection and foreshadowed the outcome. The ELISPOT assay holds promise for diagnosing pediatric IA.

2008 - BK virus infection and neurologic dysfunctions in a patient with lymphoma treated with chemotherapy and rituximab [Articolo su rivista]
Ferrari, A; Luppi, Mario; Marasca, Roberto; Potenza, Leonardo; Morselli, M; Volzone, F; Santachiara, R; Forghieri, Fabio; Barozzi, Patrizia; Torelli, Giuseppe
abstract

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2008 - Changes in the immune responses against human herpesvirus-8 in the disease course of posttransplant Kaposi sarcoma [Articolo su rivista]
Barozzi, Patrizia; Bonini, C; Potenza, Leonardo; Masetti, Michele; Cappelli, Gianni; Gruarin, P; Whitby, D; Gerunda, Giorgio Enrico; Mondino, A; Riva, Giovanni; Vallerini, Daniela; Quadrelli, Chiara; Bosco, Raffaella; Ciceri, F; Bordignon, C; Schulz, Tf; Torelli, Giuseppe; Luppi, Mario
abstract

In nine patients with posttransplant Kaposi sarcoma (KS) T-cell responses to human herpesvirus (HHV)-8 latent and lytic antigens, as detected by enzyme-linked-immunospot (Elispot) assay, were absent at disease onset. Virus-specific T-cell responses were detected in six renal recipients at remission after a reduction of calcineurin inhibitors (CIs), and in two HHV-8 seropositive renal recipients without KS. In two liver recipients undergoing switch from CIs to sirolimus (SRL), normalization of the T-cell repertoire and recovery of both HHV-8-specific effector and memory T lymphocytes were associated with complete KS remission. In a renal recipient undergoing SRL conversion, the early recovery of HHV-8-specific effector but not of memory T lymphocytes, was associated only with partial remission. Neither rejection nor changes in graft function were observed after SRL conversion. HHV-8-specific T-cell responses are required to achieve posttransplant KS remission, and may be restored under SRL, while maintaining effective immunosuppression.

2008 - HHV-6A in syncytial giant-cell hepatitis [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Barozzi, Patrizia; Rossi, Giulio; Cocchi, Stefania; Codeluppi, Mauro; Pecorari, Monica; Masetti, Michele; DI BENEDETTO, Fabrizio; Gennari, William; Portolani, Marinella; Gerunda, Giorgio Enrico; Lazzarotto, Tiziana; Landini, Maria Paola; Schulz, Thomas F.; Torelli, Giuseppe; Guaraldi, Giovanni
abstract

Syncytial giant-cell hepatitis is a rare but severe form of hepatitis that is associated with autoimmune diseases, drug reactions, and viral infections. We used serologic, molecular, and immunohistochemical methods to search for an infectious cause in a case of syncytial giant-cell hepatitis that developed in a liver-transplant recipient who had latent infection with variant B of human herpesvirus 6 (HHV-6B) and who had received the organ from a donor with variant A latent infection (HHV-6A). At the onset of the disease, the detection of HHV-6A (but not HHV-6B) DNA in plasma, in affected liver tissue, and in single micromanipulated syncytial giant cells with the use of two different polymerase-chain-reaction (PCR) assays indicated the presence of active HHV-6A infection in the patient. Expression of the HHV-6A-specific early protein, p41/38, but not of the HHV-6B-specific late protein, p101, was demonstrated only in liver syncytial giant cells in the absence of other infectious pathogens. The same markers of HHV-6A active infection were documented in serial follow-up samples from the patient and disappeared only at the resolution of syncytial giant-cell hepatitis. Neither HHV-6B DNA nor late protein was identified in the same follow-up samples from the patient. Thus, HHV-6A may be a cause of syncytial giant-cell hepatitis.

2008 - Immunoliposome for the delivery of drugs and SiRNA for the treatment of cancer [Abstract in Atti di Convegno]
Ruozi, Barbara; Riva, Giovanni; Barozzi, Patrizia; Tosi, Giovanni; Belletti, Daniela; Forni, Flavio; Luppi, Mario; Vandelli, Maria Angela
abstract

Immunoliposome for the delivery of drugs and SiRNA for the treatment of cancer

2008 - Kaposi's sarcoma after liver transplantation [Articolo su rivista]
DI BENEDETTO, Fabrizio; Di Sandro, S; De Ruvo, N; Berretta, M; Masetti, Michele; Montalti, R; Ballarin, Roberto; Cocchi, S; Potenza, Leonardo; Luppi, Mario; Gerunda, Giorgio Enrico
abstract

INTRODUCTION: Kaposi's Sarcoma (KS) is a malignant neoplasm arising from endothelial cells. HHV8-infection represents a key pathogenic determinant for the development of KS. There are no standard criteria to treat KS in immunosuppressed-individuals. Six cases (2.1%) of KS occurred in our Center among 285-recipients who underwent liver transplantation (LT) between October 2000 and November 2006. METHODS: Patients were four males and two females. Mean age was 57 years (range 44-65). Indication for LT was ESLD associated/non-associated with hepatocellular carcinoma (HCC). The immunosuppressive regimen consisted of cyclosporine/tacrolimus associated with steroids or daclizumab. HHV8-detection was performed by the serological method before LT, and by polymerase chain reaction (PCR)-analysis after KS. RESULTS: One patient had HCV-related cirrhosis and coinfection from HIV, three had HBV-related cirrhosis, two of these with coexistent HCC. The last two patients had alcoholic-cirrhosis, one with coexistent HCC. Mean time from transplantation to KS was 6.2 months (range 3.8-8.8). Three patients were treated with doxorubicin and three with switch from calcineurin-inhibitors to sirolimus. Three patients expired after 11.5, 8.8, and 7.4 months from KS diagnosis. DISCUSSION: KS should be treated by a multidisciplinary approach to obtain an early diagnosis and best management. Effective treatment with immunosuppression reduction or switch to sirolimus is mandatory and can induce complete regression.

2008 - Parvoviruses in blood donors and transplant patients, Italy [Articolo su rivista]
Vallerini, Daniela; Barozzi, Patrizia; Quadrelli, Chiara; Bosco, Raffaella; Potenza, Leonardo; Riva, Giovanni; Gregorini, G; Sandrini, S; Tironi, A; Montagnani, Giuliano; De Palma, M; Torelli, Giuseppe; Delwart, E; Luppi, Mario
abstract

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2008 - Reply to: [Efficacy of cyclosporine as a single agent therapy in chronic idiopathic thrombocytopenic purpura". Haematologica 2008; 93:e61] [Articolo su rivista]
Emilia, G.; Luppi, M.; Morselli, M.; Forghieri, F.; Potenza, L.; Torelli, G.
abstract

2008 - Veicolazione liposomiale del cidofovir nel trattamento del PEL (Primary Effusion Lynphoma) [Abstract in Rivista]
Barozzi, Patrizia; Riva, Giovanni; Ruozi, Barbara; Tosi, Giovanni; Belletti, Daniela; Potenza, Leonardo; Quadrelli, Chiara; Vallerini, Daniela; Zanetti, Eleonora; M., Morselli; Forghieri, Fabio; Maccaferri, Monica; A., Paolini; F., Volzone; Vandelli, Maria Angela; Forni, Flavio; Torelli, Giuseppe; Luppi, Mario
abstract

Veicolazione liposomiale del cidofovir nel trattamento del PEL (Primary Effusion Lynphoma)

2007 - B cells and herpesviruses: a model of lymphoproliferation [Articolo su rivista]
Barozzi, Patrizia; Potenza, Leonardo; Riva, Giovanni; Vallerini, Daniela; Quadrelli, Chiara; Bosco, Raffaella; Forghieri, Fabio; Torelli, Giuseppe; Luppi, Mario
abstract

Unlike alpha- and beta-herpesviruses, human gamma-herpesviruses, including the Epstein-Barr virus (EBV) and the human herpesvirus-8 (HHV-8), are B lymphotropic viruses. Primary infection with EBV, in otherwise healthy subjects, causes a benign lymphoproliferative syndrome, the mononucleosis syndrome. However, several epidemiologic and biologic studies have shown a pathogenetic role of EBV in the development of human B cell lymphomas, both in immunocompetent and in immunosuppressed individuals. HHV-8 is the necessary etiologic agent of a lymph vascular tumor, the Kaposi sarcoma, but it is also implicated in the pathogenesis of rare B cell lymphoproliferative disorders, mainly occurring in the setting of immunosuppression. The aim of this review is to provide an updated description of the different strategies used by these two herpesviruses to influence B cell fate decisions. Both EBV and HHV-8 have evolved specific mechanisms in order to: (1) interact with the B cell developmental machinery; (2) allow infected B cells to escape from the control of the immune system; (3) affect the B cell cycle checkpoints; (4) mimic and influence B cellular proliferation and differentiation pathways. Understanding the mechanisms of herpesvirus induced B cell lymphoproliferation will provide the basis for novel treatment approaches in patients with EBV and HHV-8 related lymphomas.

2007 - Characterization of B cell lymphoma in patients with Sjogren's syndrome and hepatitis C virus infection [Articolo su rivista]
Ramos Casals, M; La Civita, L; De Vita, S; Solans, R; Luppi, Mario; Medina, F; Caramaschi, P; Fadda, P; De Marchi, G; Lopez Guillermo, A; Font, J.
abstract

Objective. To characterize the clinical and immunologic patterns of expression, response to therapy, and outcome of patients with Sjogren's syndrome (SS) and associated hepatitis C virus (HCV) infection who developed B cell lymphoma. Methods. Various international reference centers constituted a multicenter study group with the purpose of creating a registry of patients with SS-HCV who developed B cell lymphoma. A protocol form was used to record the main characteristics of SS, chronic HCV infection, and B cell lymphoma. Results. Twenty-five patients with SS-HCV with B cell lymphoma were included in the registry. There were 22 (88%) women and 3 (12%) men (mean age 55, 58, and 61 years at SS, HCV infection, and lymphoma diagnosis, respectively). The main extraglandular SS manifestations were cutaneous vasculitis in 15 (60%) patients and peripheral neuropathy in 12 (48%); the main immunologic features were positive rheumatoid factor (RF) in 24 (96%) and type 11 cryoglobulins in 20 (80%). The main histologic subtypes were mucosa-associated lymphoid tissue (MALT) lymphoma in 11. (44%) patients, diffuse large B cell lymphoma in 6 (24%), and follicular center cell lymphoma in 6 (24%). Fifteen (60%) patients had an extranodal primary location, most frequently in the parotid gland (5 patients), liver (4 patients), and stomach (4 patients). Twelve (52%) of 23 patients died after a median followup from the time of lymphoma diagnosis of 4 years, with lymphoma progression being the most frequent cause of death. Survival differed significantly between the main types of 13 cell lymphoma. Conclusion. Patients with SS-HCV and B cell lymphoma are clinically characterized by a high frequency of parotid enlargement and vasculitis, an immunologic pattern overwhelmingly dominated by the presence of RF and mixed type 11 cryoglobulins, a predominance of MALT lymphomas, and an elevated frequency of primary extranodal involvement in organs in which HCV replicates (exocrine glands, liver, and stomach).

2007 - Cytarabine-related lung infiltrates on high resolution computerized tomography: a possible complication with benign outcome in leukemic patients. [Articolo su rivista]
Forghieri, Fabio; Luppi, Mario; Morselli, M; Potenza, Leonardo
abstract

Potentially fatal lung toxicity occurs in 12-20% of leukemic patients treated with cytarabine especially at intermediate to high doses, usually presenting as noncardiogenic pulmonary edema (NCPE). Anecdotally the association between cytarabine and the onset of bronchiolitis obliterans organizing pneumonia (BOOP) has been reported. We describe here three cases of patients affected by acute myeloid leukemia (AML) treated with chemotherapeutic regimens including high dose cytarabine, who developed early onset of fever, mild dyspnea, moderate hypoxemia on arterial blood gas analysis and lung infiltrates documented by high-resolution computerized tomography (HRCT), with a more indolent behaviour and a benign clinical outcome, compared with similar cases previously reported in the literature. Our cases widen the spectrum of clinical features of cytarabine-related toxicity in leukemic patients.

2007 - Diagnosis of invasive aspergillosis by tracking Aspergillus-specific T cells in hematologic patients with pulmonary infiltrates [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Vallerini, Daniela; Bosco, R; Quadrelli, Chiara; Morselli, M; Forghieri, Fabio; Volzone, F; Codeluppi, M; Rossi, G; Tazzioli, Giovanni; Venturelli, C; Torelli, Giuseppe; Luppi, Mario; Mediani, Laura
abstract

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2007 - ERK1/2 phosphorylation is an independent predictor of complete remission in newly diagnosed adult acute lymphoblastic leukemia [Articolo su rivista]
Gregorj, C; Ricciardi, Mr; Petrucci, Mt; Scerpa, Mc; De Cave, F; Fazi, P; Vignetti, M; Vitale, A; Mancini, M; Cimino, G; Palmieri, S; Di Raimondo, F; Specchia, G; Fabbianof, ; Cantore, N; Mosna, F; Camera, A; Luppi, Mario; Annino, L; Miraglia, E; Fioritoni, G; Ronco, F; Meloni, G; Mandelli, F; Andreeff, M; Milella, M; Foà, R; Tafuri, A; GIMEMAAcute Leukemia Working, Party
abstract

Extracellular signal-regulated kinase-1/2 (ERK1/2) is frequently found constitutively activated (p-ERK1/2) in hematopoietic diseases, suggesting a role in leukemogenesis. The aim of this study was to assess the expression and clinical role of p-ERK1/2 in adult acute lymphoblastic leukemia (ALL). In 131 primary samples from adult de novo ALL patients enrolled in the Gruppo Italiano per le Malattie Ematologiche dell'Adulto (GIMEMA) Leucemia Acute Linfoide (LAL) 2000 protocol and evaluated by flow cytometry, constitutive ERK1/2 activation was found in 34.5% of cases; these results were significantly associated with higher white blood cell (WBC) values (P=.013). In a multivariate analysis, p-ERK1/2 expression was an independent predictor of complete remission achievement (P=.027). Effective approaches toward MEK inhibition need to be explored in order to evaluate whether this may represent a new therapeutic strategy for adult ALL patients.

2007 - Epstein-Barr virusassociated pneumonia in an adult patient with severe aplastic anaemia: resolutionafter the transient withdrawal of cyclosporine. [Articolo su rivista]
Potenza, Leonardo; Barozzi, Patrizia; Codeluppi, M; Morselli, M; Forghieri, Fabio; Volzone, F; Riva, Giovanni; Pietrosemoli, P; Pecorari, M; Leonardi, G; Torelli, Giuseppe; Luppi, Mario
abstract

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2007 - Fatal hemophagocytic syndrome related to active human herpesvirus-8/Kaposi sarcoma-associated herpesvirus infection in human immunodeficiency virus-negative, non-transplant patients without related malignancies [Articolo su rivista]
Re, A; Facchetti, F; Borlenghi, E; Cattaneo, C; Capucci, Ma; Ungari, M; Barozzi, Patrizia; Vallerini, Daniela; Potenza, Leonardo; Torelli, Giuseppe; Rossi, G; Luppi, Mario
abstract

Hemophagocytic syndrome (HS) may occur as a consequence of herpes viral infections. Human herpesvirus 8 (HHV-8)/Kaposi sarcoma-associated herpesvirus has so far been recognized as a trigger of HS only in immunosuppressed subjects or in patients with Kaposi sarcoma and/or HHV-8-related lymphoproliferative diseases. We report two Italian human immunodeficiency virus (HIV)-negative elderly men who developed an HS with a rapidly fatal course, following treatment with corticosteroids for autoimmune hemolytic anemia. An overwhelming active infection with HHV-8 was unequivocally documented by molecular and immunohistochemical methods, in the absence of HHV-8-related tumors. The occurrence of HHV-8-associated HS, although rare, may be considered, even out of the HIV or the transplantation settings, at least in areas endemic for HHV-8 infection.

2007 - Helicobacter pylori infection andchronic immune thrombocytopenic purpura: long-term results of bacteriumeradication and association with bacterium virulence profiles [Articolo su rivista]
Emilia, Giovanni; Luppi, Mario; Zucchini, Patrizia; Morselli, M; Potenza, Leonardo; Forghieri, Fabio; Volzone, F; Jovic, Gordana; Leonardi, G; Donelli, A; Torelli, Giuseppe
abstract

Eradication of Helicobacter pylori may lead to improvement of chronic immune thrombocytopenic purpura (ITP), although its efficacy over time is uncertain. We report the results of H pylori screening and eradication in 75 consecutive adult patients with ITP. We also used molecular methods to investigate lymphocyte clonality and H pylori genotypes in the gastric biopsies from 10 H pylori-positive patients with ITP and 19 H pylori-positive patients without ITP with chronic gastritis. Active H pylori infection was documented in 38 (51%) patients and successfully eradicated in 34 (89%) patients. After a median follow-up of 60 months, a persistent platelet response in 23 (68%) of patients with eradicated infection was observed; 1 relapse occurred. No differences in mucosal B- or T-cell clonalities were observed between patients with ITP and control participants. Of note, the frequency of the H pylori cagA gene (P = .02) and the frequency of concomitant H pylori cagA, vacAs1, and iceA genes (triple-positive strains; P = .015) resulted statistically higher in patients with ITP than in control participants. All asymptomatic H pylori-positive patients with ITP were suffering from chronic gastritis. Our data suggest a sustained platelet recovery in a proportion of patients with ITP by H pylori eradication alone. Overrepresentation of specific H pylori genotypes in ITP suggests a possible role for bacterium-related factors in the disease pathogenesis.

2007 - May the correlation between Kit-D816 mutation and AML1-ETO level change the use of prognostic factors in t(8;21) AML? [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Riva, Giovanni; Zucchini, Patrizia; Morselli, M; Volzone, F; Forghieri, Fabio; Torelli, Giuseppe; Ottaviani, E; Martinelli, G.
abstract

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2006 - A non-chemotherapy treatment of a primary effusion lymphoma: durable remission after intracavitary cidofovir in HIV negative PEL refractory to chemotherapy [Articolo su rivista]
Halfdanarson, Tr; Markovic, Sn; Kalokhe, U; Luppi, Mario
abstract

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2006 - Diagnosis of occult tuberculosis in hematological malignancy by enumeration of antigen-specific T cells. [Articolo su rivista]
Richeldi, Luca; Luppi, Mario; Losi, Monica; Luppi, Fabrizio; Potenza, Leonardo; Roversi, P; Cerri, Stefania; Millington, Ka; Ewer, K; Fabbri, Leonardo; Torelli, Giuseppe; Lalvani, A.
abstract

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2006 - Giant cell hepatitis following primary infection with HHV-6 variant A, transmitted from the donor, in a liver transplant recipient latently infected with HHV-6 variant B [Abstract in Rivista]
Guaraldi, Giovanni; Cocchi, S; Codeluppi, M; Pecorari, M; Barozzi, P; Gennari, W; Bagni, A; Bosco, R; Vallerini, D; DI BENEDETTO, Fabrizio; Masetti, Michele; Portolani, M; Torelli, G; Luppi, Mario
abstract

Syncytial giant-cell hepatitis is a rare but severe form of hepatitis that is associated with autoimmune diseases, drug reactions, and viral infections. We used serologic, molecular, and immunohistochemical methods to search for an infectious cause in a case of syncytial giant-cell hepatitis that developed in a liver-transplant recipient who had latent infection with variant B of human herpesvirus 6 (HHV-6B) and who had received the organ from a donor with variant A latent infection (HHV-6A).

2006 - Human herpesvirus 6 latency characterized by high viral load: Chromosomal integration in many, but not all, cells [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Bosco, Raffaella; Vallerini, Daniela; Potenza, Leonardo; Forghieri, Fabio; Torelli, Giuseppe
abstract

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2006 - Isolated extramedullary relapse after autologous bone marrow transplantation for acute myeloid leukemia: Case report and review of the literature [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Riva, Giovanni; M., Morselli; Ferrari, Alberto; Imovilli, Annalisa; F., Giacobbi; Temperani, Paola; A., Donelli; Narni, Franco; Torelli, Giuseppe
abstract

Isolated extramedullary relapse (IEMR) is a pattern of acute myeloid leukemia (AML) relapse post-allogeneic bone marrow transplantation (alloBMT). Less is known about IEMR post-autologous BMT (autoBMT) and about factors associated with IEMR. We report a case of a woman with M4 AML who experienced IEMR post-autoBMT and review the related literature. Seventy-two alloBMT and 3 autoBMT patients, including ours, were identified. The review suggests that an M2 or M4 French-American-British (FAB) phenotype, intermediate cytogenetic risk group, and chromosome 8 abnormalities are more frequently associated with the occurrence of IEMR. IEMR occurs earlier in autoBMT than in alloBMT. Combined treatment with radiation and high-dose chemotherapy may be effective. When we searched the European Bone Marrow Transplant Registry (EBMTR) database, we found the incidence of IEMR to be statistically greater in alloBMT than in autoBMT (11% vs. 6%; P = 0.02), but no correlations have been found with the conditioning transplant regimen used. A closer follow-up, including body and central nervous system scan, should be considered in patients who are undergoing BMT presenting with several IEMR-associated factors.

2006 - KSHV/HHV-8 infection of tubular epithelial cells in transplantation kidney [Articolo su rivista]
Barozzi, Patrizia; R., Bosco; Vallerini, Daniela; Potenza, Leonardo; Torelli, Giuseppe; Luppi, Mario; F., Facchetti; Guaraldi, Giovanni; T. F., Schulz
abstract

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2006 - Kaposi's sarcoma triggered by endogenous HHV-8 reactivation after non-myeloablative allogeneic haematopoietic transplantation [Articolo su rivista]
Bruno, B; Sorasio, R; Barozzi, Patrizia; Vieira, J; Omede, P; Giaretta, F; Rotta, M; Giaccone, L; Massaia, M; Luppi, Mario; Boccadoro, M.
abstract

Human herpesvirus 8 (HHV-8) is causally associated with Kaposi's sarcoma (KS). KS is most frequently observed in HIV patients and in solid organ transplant recipients. The role of HHV-8 in allogeneic haematopoietic cell transplantation (HCT) remains to be determined. Here we describe a case in which KS concomitantly occurred with CMV reactivation after a non-myeloablative allogeneic HCT and presented with skin lesions, but not visceral involvement. Skin biopsy confirmed the diagnosis and ruled out graft versus host disease or disease recurrence. Molecular findings indicated viral reactivation of the recipient's primary infection. Tumour lesions completely receded when immunosuppression was tapered. Prevalence studies in donors and recipients are needed to determine the clinical impact of HHV-8 in HCT.

2006 - More on: Platelet count and the Use of Recombinant Factor VIIa for the treatment of Bleeding Complications after Hematopoietic Stem Cell Transplantation [Articolo su rivista]
Marietta, M; Facchini, L; Girardis, Massimo; Luppi, Mario; Torelli, Giuseppe
abstract

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2006 - Ocular involvement as first sign of isolated CNS relapse in diffuse large B-cell lymphoma [Articolo su rivista]
A., Ferrari; Luppi, Mario; A., Lazzerini; Potenza, Leonardo; Cavallini, Gian Maria; Torelli, Giuseppe
abstract

A 52-year-old man was diagnosed with stage IVB non-Hodgkin lymphoma of diffuse large B-cell lymphoma subtype that involved the bone marrow and liver. Analysis of serum samples showed a raised concentration of lactate dehydrogenase, suggesting a high–intermediate risk of non-Hodgkin lymphoma according to the international prognostic index, but no HIV-1. The patient received six cycles of CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) plus rituximab, and achieved complete remission. 6 months after diagnosis and 2 months after complete remission was achieved, the patient was admitted to hospital because of loss of vision. In the early phase of the examination, a fluorescein angiogram of the right eye (figure A) showed two large hyperfluorescent infiltrates with granularity (arrows) with small regions of retinal pigment epithelial detachments (arrowheads). In the late phase of the study, the left eye (figure B) showed a diffuse retinal pigment epithelial detachment that remained densely hypofluorescent (arrows), suggesting presence of lymphoma. However, bone-marrow biopsy and a CT scan of the neck, chest, and abdomen showed no relapse of lymphoma. 6 days later, the patient became somnolent and agitated, and developed confusion and lethargy. A CT scan (not shown) and an MRI of the brain (figure C) showed multifocal intraparenchymal lesions. Histological and immunohistochemical analysis of a stereotactic biopsy sample of one of the frontal lesions showed presence of B-lymphocyte antigen CD20-positive diffuse large B-cell lymphoma, and no presence of Epstein-Barr virus. The patient was given high-dose methotrexate followed by whole-brain radiotherapy. The patient died 1 month after CNS relapse because of disease progression.

2005 - A comprehensive genetic classification of adult acute lymphoblastic leukemia (ALL): Analysis of the GIMEMA 0496 protocol [Articolo su rivista]
Mancini, M; Scappaticci, D; Cimino, G; Nanni, M; Derme, V; Elia, L; Tafuri, A; Vignetti, M; Vitale, A; Cuneo, A; Castoldi, G; Saglio, G; Pane, F; Mecucci, C; Camera, A; Specchia, G; Tedeschi, A; Di Raimondo, F; Fioritoni, G; Fabbiano, F; Marmont, F; Ferrara, F; Cascavilla, N; Todeschini, G; Nobile, F; Kropp, Mg; Leoni, P; Tabilio, A; Luppi, Mario; Annino, L; Mandelli, F; Foà, R.
abstract

The Gruppo Italiano Malattie Ematologiche dell'Adulto (GIMEMA) 0496 protocol, through the central handling of bone marrow samples at presentation, allowed us to combine cytogenetic and molecular information on a large series of adults with acute lymphoblastic leukemia (ALL) treated homogeneously, enabling us to define as broadly as possible their genetic profile and to determine the impact on outcome of the cytogenetic-molecular signature. Of 414 patients centrally processed, 325 were considered for the categorization into the following cytogenetic-molecular subgroups: normal, t(9;22)/BCR-ABL, t(4;11)/MLL-AF4, t(1;19)/E2A-PBX1, 9p/p15-p16 deletions, 6q deletions, miscellaneous structural abnormalities, and hyperdiploid. The inclusion into each subgroup was based on a hierarchical approach: molecular abnormalities with adverse prognosis had precedence over karyotypic changes with less-defined prognosis and the latter over ploidy. Patients without abnormalities and those with isolated 9p/p15-p16 deletions showed a relatively favorable outcome (median disease-free survival [DFS], > 3 years). The t(9;22)/BCR-ABL, t(4;11)/MLL-AF4, t(1; 19)/E2A-PBX1 defined a group with dismal prognosis (median DFS, 7 months), whereas 6q deletions, miscellaneous aberrations, and hyperdiploidy predicted an intermediate prognosis (median DFS, 19 months). This study highlights the importance of a combined cytogenetic-molecular profiling of adult ALL at presentation as a critical independent determinant of their outcome, providing further evidence of the necessity of a risk-adapted therapeutic algorithm for an optimal management of these patients.

2005 - Cytomegalovirus and ClostridiumDifficile co-infection in severe ulcero-hemorrhagic colitis during inductionchemotherapy for acute lymphoblastic leukemia [Articolo su rivista]
Riva, Giovanni; Luppi, Mario; Potenza, Leonardo; Morselli, M; Ferrari, A; Saviola, Alessia; Volzone, F; Imovilli, Annalisa; Merighi, A; Maiorana, Antonino; Torelli, Giuseppe
abstract

Here we describe the first case of a biopsyprovenCytomegalovirus ulcero-hemorrhagic colitis,associated with Clostridium Difficile co-infection,occurring during standard induction chemotherapyfor common B cell acute lymphoblastic leukemia.We discuss the case and focalize clinical managementand diagnostic issues arising from it.

2005 - Efficacy of imatinib mesylate as maintenance therapy in adults with acute lymphoblastic leukemia in first complete remission [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Riva, Giovanni; Marasca, Roberto; Martinelli, Silvia; Torelli, Giuseppe
abstract

Seven Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL) patients in first complete remission received maintenance therapy with imatinib alone. Two-year progression-free survival was 75%. Quantitative polymerase-chain-reaction (qPCR) monitoring of BCR-ABL showed that: (i) persisting molecular complete response (CR) was associated with long-lasting CR; (ii) molecular relapse did not invariably mean hematologic relapse; (iii) only the wide and rapid increment of BCR-ABL values was predictive of leukemia relapse.

2005 - Herpes simplex virus pneumonia during standard induction chemotherapy for acute leukemia: case report and review of literature. [Articolo su rivista]
Ferrari, A.; Luppi, Mario; Potenza, Leonardo; Riva, Giovanni; Morselli, M.; Imovilli, A.; Volzone, F.; Rossi, G.; Codeluppi, M.; Guaraldi, Giovanni; Torelli, Giuseppe
abstract

Reactivation of latent HSV is the most common viral infection in patients during the profound neutropenia that occurs during remission induction in patients with lymphoma and acute leukemia and during the conditioning phase of bone marrow transplantation. We report here the occurrence of HSV-1 pneumonia in a patient with B-cell common ALL.

2005 - Human Cytomegalovirus, Human Herpesvirus 8 and Other Herpesviruses (Part III, Section 2, Chapter 75) [Capitolo/Saggio]
Maciejewski, J. P.; Luppi, Mario; Torelli, Giuseppe
abstract

Written by a young, innovative author group, this exciting new book gives you the clinically relevant aspects of hematology in an innovative, unique format. The book is structured to comprehensively cover the complete scope of hematology, but allows fast access to key information you need in everyday practice. A section on consultative hematology includes chapters on special populations (pregnancy, pediatrics, geriatrics), infections of marrow and blood, and hematologic problems of medical practice and surgery. You'll also find a section on tools for the hematologist, covering clinical aspects of transfusion, transplantation, and the latest innovative laboratory procedures.

2005 - Infectious agents and human immune diseases: Lessons from Helicobacter pylori [Articolo su rivista]
Emilia, G; Luppi, Mario; Torelli, Giuseppe
abstract

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2005 - Pneumonia in acute leukaemia:blossoming culprits [Articolo su rivista]
Potenza, Leonardo; Riva, Giovanni; Luppi, Mario; Torelli, Giuseppe
abstract

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2005 - Successful treatment with liposomal doxorubicin and foscarnet in a patient with widespread Kaposi's sarcoma and human herpes virus 8-related, serioushemophagocytic syndrome, after renal transplantation [Articolo su rivista]
Bossini, N; Sandrini, S; Setti, G; Luppi, Mario; Maiorca, P; Maffei, C; Cancarini, G.
abstract

BACKGROUND: It is well known that the human herpes virus 8 (HHV8) is linked to several malignancies such as Kaposi's sarcoma (KS). Moreover, pancytopenia due to hemophagocytic syndrome could be associated with HHV8 infection. In renal transplant recipients affected by KS, the tapering of immunosuppression often leads to KS remission, but also results in graft loss in >50% of cases. Chemotherapy and antiviral therapy have also been used, mainly in the presence of visceral involvement. CASE REPORT: We describe a transplant recipient with widespread cutaneous and visceral KS HHV8 associated, complicated by hemophagocytic syndrome. At transplantation the patient's serology for HHV8 was negative, but thereafter it became positive. The first step in treatment (cyclosporine dose reduction until suspension) failed to improve the clinical course. Therefore, therapy combining liposomal doxorubicin and foscarnet was started. Clearance of HHV8 in the blood and complete resolution of the KS lesions were achieved. Immunosuppression with cyclosporine was resumed. No KS relapse has occurred, blood tests for HHV8 are negative, and graft function is good after a 5-yr follow-up. CONCLUSIONS: Therapy combining liposomal doxorubicin and foscarnet was effective in this renal transplant recipient with KS and HHV8 infection and enabled us to resume immunosuppressive therapy; therefore, reducing the risk of acute/chronic rejection.

2005 - Suppressive effect of human herpesvirus-8/Kaposi sarcoma-associated herpesvirus on in vitro colony formation of hematopoietic progenitor cells [Articolo su rivista]
Luppi, Mario; Trovato, R; Barozzi, Patrizia; Gibellini, F; Potenza, Leonardo; Riva, Giovanni; Torelli, Giuseppe
abstract

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2005 - Treatment of herpesvirus associated primary effusion lymphoma with intracavity cidofovir [Articolo su rivista]
Luppi, Mario; Trovato, R; Barozzi, Patrizia; Vallisa, D; Rossi, Giorgio; Re, A; Ravazzini, L; Potenza, Leonardo; Riva, Giovanni; Morselli, M; Longo, G; Cavanna, L; Roncaglia, R; Torelli, Giuseppe
abstract

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2004 - A t(11;20)(p15;q11) may identify a subset of nontherapy-related acute myelocytic leukemia [Articolo su rivista]
Potenza, Leonardo; B., Sinigaglia; Luppi, Mario; M., Morselli; A., Saviola; A., Ferrari; Riva, Giovanni; Zucchini, Patrizia; F., Giacobbi; G., Emilia; Temperani, Paola; Torelli, Giuseppe
abstract

A t(11;20)(p15;q11) is a rare but recurrent chromosomal aberration, reported in one case of polycythemia vera and a few cases of de novo acute myelocytic leukemia (AML) and therapy-related myelodysplastic syndrome (t-MDS). In t-MDS cases, the translocation resulted in the NUP98/TOP1 fusion transcript. The NUP98 gene has been suggested as the target for therapy-related malignancies. The reciprocal TOP1/NUP98 chimera, however, has not yet been encountered. We report a further case of de novo AML, subtype M2 in the French-American-British (FAB) classification, in which the reverse-transcriptase polymerase chain reaction (RT-PCR) revealed the NUP98/TOP1 chimera and also, for the first time, its reciprocal TOP1/NUP98. The literature review disclosed that, among six cases of de novo AML with t(11;20), the NUP98 gene was shown to be involved in one case and the NUP98/TOP1 chimera was detected in another. The translocation seems to be frequently associated with the FAB M2 subtype, younger age, hyperleukocytosis, and poor prognosis; thus, this translocation may identify a subset of not-therapy-related AML patients with shared clinical features.

2004 - AUTOLOGOUS STEM CELL TRANSPLANTATION FOR NEWLY DIAGNOSED FOLLICULAR LYMPHOMAS: LOW TOXICITY AND EXTENDED PROGRESSION FREE SURVIVAL. R1235 Barcelona, Spain. Bone Marrow Transplantation March 2004, Volume 33, Supplement 1 [Abstract in Rivista]
Narni, Franco; A., Donelli; A., Cuoghi; P., Bresciani; Pozzi, Samantha; Marasca, Roberto; G., Leonardi; M., Morselli; Luppi, Mario; G., Longo; Torelli, Giuseppe
abstract

In spite of an indolent course, advanced stage follicularlymphomas (FL) are incurable with conventional chemotherapy.High dose therapy could be investigated in selected patients,provided toxicity were acceptable. Improvements in supporttherapy and the widespread use of peripheral blood stem cells(SC) have made autologous transplantation a relatively safeprocedure. This prompted us to start a pilot study and, since1995, 23 patients have been enrolled at our center. Median agewas 49 (26 - 65). Most patients (80%) had stage IV disease andbone marrow involvement. For patients who achieved a completeresponse or a good partial response after CHOP, SC weremobilized with cyclophosphamide or DHAP plus G-CSF, andhigh dose therapy consisted of BEAM. High risk patients or poorresponders (residual high tumor burden or bone marrowinfiltration >50%) switched to a program of high dose sequentialtherapy. Nine patients received not-manipulated hematopoieticSC, 7 received ''ex vivo'' purged SC (6 by CD34+ positiveselection; 1 by positive+negative selection), and 8 patientsunderwent ''in vivo'' purging with anti-CD20 monoclonal antibody(rituximab). Residual disease was monitored at different steps bynested PCR. Treatment was well tolerated, with a median time toengrafment of 9 days (7-12), and a median stay in hospital of 22days (15-35). RBC and platelet transfusions were necessary,respectively, in 52% and 100% of the patients. The meannumber of RBC and platelet units given to transfused patientswas, respectively, 2.2 (1-5) and 3.7 (1-7). Fever occurred in 52%of the patients (mean 2,8 days in febrile patients) and mucositisin 85%. We did not observe, so far, any case of myelodysplasiaor secondary cancer. With a median FU of 58 (8-95) monthsfrom transplant, all patients are alive, 5 have relapsed, and 5-years estimated PFS is 70%. CD34+ selection had no impact onrelapse. In one patient, molecular response could be restoredand maintained by rituximab alone. Only a few data have beenpublished regarding autologous transplantation in FL patients atdiagnosis. Although the FU is relatively short and the number ofpatients still small, the present data suggest that autologoustransplant is a safe and effective therapy for selected, not heavilypretreated young patients with advanced stage disease.Especially since ''in vivo'' purging strategies can easily be carriedout, autologous SC transplantation should be investigated inrandomized trials.

2004 - Autologous stem cell transplantation for newly diagnosed follicular lymphomas: low toxicity and extended progression free survival [Articolo su rivista]
Narni, Franco; Donelli, A; Cuoghi, A; Bresciani, P; Pozzi, S; Marasca, Roberto; Leonardi, G; Morselli, M; Luppi, Mario; Longo, G; Torelli, Giuseppe
abstract

2004 - Cardiac involvement in malignancies. Case 2. Right ventricular lesion as presenting feature of acute promyelocytic leukemia [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Morselli, M; Riva, Giovanni; Saviola, A; Ferrari, Alberto; De Santis, M; Rossi, Rosario; Torelli, Giuseppe
abstract

N/A

2004 - Gemtuzumab ozogamicin for relapsed and refractory acute myeloid leukemia and myeloid sarcomas [Articolo su rivista]
Piccaluga, Pp; Martinelli, G; Rondoni, M; Malagola, M; Gaitani, S; Isidori, A; Bonini, A; Gugliotta, L; Luppi, Mario; Morselli, M; Sparaventi, G; Visani, G; Baccarani, M.
abstract

Antibody-targeted chemotherapy is a promising approach in patients with hematological malignancies. In particular, gemtuzumab ozogamicin (GO, formerly CMA-676), an anti-CD33 antibody linked to calicheamicin, has been approved for the treatment of elderly patients with acute myeloid leukemia (AML) in relapse. Nevertheless, no data are until now available concerning the possible efficacy of GO for myeloid sarcomas ( MS). We treated with GO 24 AML patients, in 5 cases presenting with myeloid sarcomas of the skin or bones. The overall complete response rate was 21%. The median duration of response was 6 months. Four out of the 5 patients with myeloid sarcoma showed a regression of the masses, in two cases also obtaining a clearance of marrow blasts. The most common adverse events included thrombocytopenia, neutropenia, infections, elevation of bilirubin and hepatic transaminases. Notably, severe bleeding occurred in 5 cases ( 21%). VOD was documented in 1 case. We conclude that GO is effective as a single agent in AML and myeloid sarcomas. Further data are required to clarify the possible correlation between GO administration and occurrence of bleeding.

2004 - Helicobacter pylori and idiopathic thrombocytopenic purpura [Articolo su rivista]
Emilia, G; Luppi, Mario; Torelli, Giuseppe
abstract

n.d.

2004 - Is it now the time to update treatment protocols for lymphomas with new anti-virus systems? [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Potenza, Leonardo; Riva, Giovanni; Morselli, M; Torelli, Giuseppe
abstract

n.d.

2004 - MALT Lymphomas: Epidemiology and Infectious Agents [Capitolo/Saggio]
Luppi, Mario; R., Negrini
abstract

In 1983 Isaacson and Wright observed that low grade lymphomas arising from the stomach, thyroid, salivary glands and lung exhibit histological features that reproduce the architecture of mucosa associated lymphoid tissue (MALT) rather than that of similar lymph node tumor. The tumor cells are located in the marginal zone surrounding the mantle, and like the B cells of the marginal zone of Peyer\'s patches and tonsils, invade the overlying epithelium, forming the so-called “lymphoepithelial lesion“; like normal MALT, these lymphomas have reactive lymphoid follicles. MALT lymphomas have been defined as extranodal marginal zone B-cell lymphomas of MALT type both in the REAL classification and in the last WHO Classification. These tumors present an indolent natural course, since they tend to remain localized for long periods and show a good response to the therapy. They usually arise from organs where MALT is physiologically absent and is acquired following a persistent antigenic stimulation by an infectious agent or by an autoimmune process.

2004 - Myocardial ischemia in a patient with acute lymphoblastic leukemia during L-asparaginase therapy [Articolo su rivista]
Saviola, A; Luppi, Mario; Potenza, Leonardo; Morselli, M; Bresciani, P; Ferrari, Alberto; Riva, Giovanni; Torelli, Giuseppe
abstract

Haemostatic abnormalities may occur in 1-2% of patients treated with L-asparaginase. Here, we present the second case of a myocardial infarction, developing in a patient with acute lymphoblastic leukemia (ALL), in the course of L-asparaginase treatment. In our patient and in the only one reported case from the literature, a recent exposure to vincristine and daunorubicin was also reported, but induction chemotherapy program was completed as scheduled, with the only withdrawal of L-asparaginase. Myocardial infarction should be included in the list of thrombotic complications possibly associated with L-asparaginase treatment, or with a combination of L-asparaginase and vinca alkaloids/anthracycline.

2004 - Thrombotic complications associated with thalidomide in multiple myeloma: an old problem with new questions and quandaries in decision-making [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Morselli, M; Saviola, A; Ferrari, Alberto; Riva, Giovanni; Longo, G; Marietta, M; Torelli, Giuseppe
abstract

n.d.

2003 - Acquired haemophilia in HIV negative, HHV-8 positive multicentric Castleman's disease: a case report [Articolo su rivista]
M., Marietta; Pozzi, Samantha; Luppi, Mario; M., Bertesi; C., Cappi; M., Morselli; Torelli, Giuseppe
abstract

Multicentric Castleman's Disease (MCD) is an atypical lymphoproliferative disorder, related to human herpesvirus 8 (HHV-8) infection and often associated with autoimmune diseases such as haemolytic anaemia and thrombocytopenia. Acquired haemophilia (AH) is a rare, life-threatening disease, which can occur in association with lymphoproliferative disorders, although only one case of AH in MCD has been described so far. We report the case of a human immuno deficiency virus negative 71-yr-old woman referred to our hospital for prolonged bleeding on surgical site following a lymph node biopsy. Lymph node histology revealed MCD, while the screening for the bleeding disorder showed prolonged activated partial thromboplastin time (APTT) (ratio: 1.89, normal value <1.24), low factor VIII (FVIII:C) levels (6%) with anti-factor VIII antibodies (2.3 Bethesda units), leading to a diagnosis of AH. Virological studies on plasma, lymphocyte and bronchoalveolar wash showed positivity for HHV-8 infection. Treatment with steroids (metilprednisolone 1-1.5 mg/kg/d) and cyclophosphamide (100 mg/d orally) was unsuccessful, and then antiviral therapy with cidofovir (5 mg/kg/wk) was started. A transient normalisation of APTT was seen after two administrations of cidofovir, but then coagulation parameters worsened and a large haematoma of the arm appeared. Bleeding was successfully stopped with two boluses of recombinant activated factor VII (Novoseven 90 microg/kg). Therapy with anti-CD 20 monoclonal antibody rituximab (Mabthera 375 mg/m2 once a week for 4 wk) was started, and following two administrations APTT normalised once again. Cardiological and neurological complications arose before the third dose of rituximab and the patient died shortly afterwards.

2003 - Erratum: Post-transplant Kaposi sarcoma originates from the seeding of donor-derived progenitors (Nature Medicine (2003) 9 (554-561)) [Articolo su rivista]
Barozzi, P.; Luppi, M.; Facchetti, F.; Mecucci, C.; Alu, M.; Sarid, R.; Rasini, V.; Ravazzini, L.; Rossi, E.; Festa, S.; Crescenzi, B.; Wolf, D. G.; Schulz, T. F.; Torelli, G.
abstract

2003 - Human herpesvirus 8-associated diseases in solid-organ transplantation: importance of viral transmission from the donor [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Guaraldi, Giovanni; L., Ravazzini; Rasini, Valeria; C., Spano; Riva, Giovanni; Vallerini, Daniela; Ad, Pinna; Torelli, Giuseppe
abstract

No abstract available

2003 - Immune thrombocytopenic purpura and Helicobacter pylori infection [Articolo su rivista]
M., Morselli; Potenza, Leonardo; Luppi, Mario; Torelli, Giuseppe; G., Emilia
abstract

No abstract available

2003 - KSHV reactivation in post-transplant Kaposi sarcoma - Reply [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Facchetti, F; Schulz, Tf; Torelli, Giuseppe
abstract

No Abstract available

2003 - Late occurrence of hepatic veno-occlusive disease following gemtuzumab ozogamicin: successful treatment with defibrotide [Articolo su rivista]
Saviola, A; Luppi, Mario; Potenza, Leonardo; Morselli, M; Ferrari, Alberto; Riva, Giovanni; Torelli, Giuseppe
abstract

No abstract available

2003 - Leukaemic pulmonary infiltrates in adult acute myeloid leukaemia: a high-resolution computerized tomography study [Articolo su rivista]
Potenza, Leonardo; Luppi, Mario; Morselli, M; Tonelli, S; D'Apollo, N; Facchini, L; Torricelli, Pietro; Tazzioli, Giovanni; Saviola, A; Bresciani, P; Longo, G; Torelli, Giuseppe
abstract

Leukaemic infiltration of the lungs may occur in acute myeloid leukaemia (AML). Pulmonary infiltrates are usually microscopic and invariably associated with hyperleucocytosis. Four AML patients with respiratory symptoms and low leucocyte counts underwent standard chest radiography, bronchoscopy with bronchoalveolar lavage and high-resolution computerized tomography (HRCT) of the lungs. HRCT scans showed pulmonary infiltrates with alveolar, interstitial, mixed and peribronchial/perivascular patterns in all patients, including one with negative standard radiographic findings. Infectious agents were excluded. Histology of the lung biopsy/autopsy specimens showed leukaemic infiltrates. Pulmonary leukaemia may be the cause of pulmonary infiltrates, even in non-hyperleucocytosic AML patients with low blast counts.

2003 - Mysticism to medicine [Articolo su rivista]
Barozzi, Patrizia; Luppi, Mario; Torelli, Giuseppe
abstract

No abstract available

2003 - Necrotising dermatitis in refractory acute myeloid leukaemia [Articolo su rivista]
D'Apollo, N; Saviola, A; Longo, G; Luppi, Mario; Emilia, G; Torelli, Giuseppe
abstract

Severe cutaneous infections in leukaemic patients are difficult to treat and can rapidly become fatal. We report on a case of essential thrombocythemia evolved to a myelodysplastic syndrome and finally, to an overt myeloid leukaemia, refractory to chemotherapy. In the presence of a marked neutropenia, the patients developed a wide Staphylococcus epidermidis necrotising dermatitis. The diagnosis was made possible only by a skin biopsy culture and the antibiotic treatment, based on antimicrobial susceptibility tests, rapidly resolved the infection. In neutropenic patients, appropriate laboratory tests and treatment, can lead to recovery of life-threatening infections.

2003 - Post-transplant Kaposi sarcoma originates from the seeding of donor-derived progenitors [Articolo su rivista]
Barozzi, Patrizia; Luppi, Mario; F., Facchetti; C., Mecucci; Alu, M.; R., Sarid; V., Rasini; L., Ravazzini; E., Rossi; S., Festa; B., Crescenzi; Dg, Wolf; Tf, Schulz; Torelli, Giuseppe
abstract

Kaposi sarcoma (KS) is a vascular tumor that can develop in recipients of solid tissue transplants as a result of either primary infection or reactivation of a gammaherpesvirus, the KS-associated herpesvirus, also known as human herpesvirus-8 (HHV-8). We studied whether HHV-8 and the elusive KS progenitor cells could be transmitted from the donor through the grafts. We used a variety of molecular, cytogenetic, immunohistochemical and immunofluorescence methods to show that the HHV-8-infected neoplastic cells in post-transplant KS from five of eight renal transplant patients harbored either genetic or antigenic markers of their matched donors. These data suggest the use of donor-derived HHV-8-specific T cells for the control of post-transplant KS.

2003 - Skin cancers after organ transplantation [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Torelli, Giuseppe
abstract

No abstract available

2003 - Unsuccessful treatment with voriconazole of a brain abscess due to Cladophialophora bantiana [Articolo su rivista]
Fica, A.; Diaz, M. -C.; Luppi, M.; Olivares, R.; Saez, L.; Baboor, M.; Vasquez, P.
abstract

Cladophialophora bantiana is a dematiaceous fungus, associated with brain abscess in normal or immunosuppressed patients. We report a case of CNS infection in this agent unsuccessfully treated by surgery and various antifungal compounds including high doses of voriconazole (6 mg/kg bid). No adverse effects related to this compound were observed.

2002 - 'Gloves and socks' papular purpuric syndrome following primary infection with parvovirus B19: a link between dermatologists and haematologists [Articolo su rivista]
Tonelli, S; Luppi, Mario; Morselli, M; Facchini, L; Potenza, Leonardo; Torelli, Giuseppe
abstract

No abstract available

2002 - HHV-8 infection in the transplantation setting: A concern only for solid organ transplant patients? [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Rasini, Valeria; Torelli, Giuseppe
abstract

Early epidemiologic studies have shown an increased risk of Kaposi sarcoma (KS) in recipients of solid organ transplants, while KS is exceptional in the setting of autologous and allogeneic bone marrow (BM) and peripheral blood stem cell (PBSC) transplant patients. The recent discovery of human herpesvirus 8 (HHV-8) as the necessary etiologic agent of KS has stimulated studies to assess whether KS is the result of HHV-8 transmission from the donor or of reactivation of a pre-existing HHV-8 infection in the recipient host. An association of HHV-8 infection with lymphoid neoplasias has also been observed, identifying this herpesvirus as a possible causal agent of some Epstein-Barr virus negative post-transplant lymphoproliferative diseases. The recent description of non-neoplastic complications associated with HHV-8 reactivation after autologous PBSC transplantation, has raised concerns about the spectrum of diseases potentially associated with this herpesvirus, even in the setting of BM and or PBSC transplantation. The issue of HHV-8 transmission with the grafts, the problems inherent with the diagnosis and monitoring of active viral infection, the need for prevention and treatment of the clinical consequences of HHV-8 primary infection and reactivation cannot be underestimated, at least in areas endemic for HHV-8 infection.

2002 - Helicobacter pylori infection and idiopathic thrombocytopenic purpura [Articolo su rivista]
Emilia, G; Luppi, Mario; Morselli, M; Potenza, Leonardo; D'Apollo, N; Torelli, Giuseppe
abstract

No abstract available

2002 - Human herpesvirus 8-associated primary effusion lymphoma in human immunodeficiency virus-negative patients: a clinico-epidemiologic variant resembling classic Kaposi's sarcoma [Articolo su rivista]
Ascoli, V; Lo Coco, F; Torelli, Giuseppe; Vallisa, D; Cavanna, L; Bergonzi, C; Luppi, Mario
abstract

No abstract available

2002 - Human herpesvirus-8 and HIV [Capitolo/Saggio]
Biberfeld, P; Lebbe, C; Tschachler, E; Luppi, Mario
abstract

No Abstract available

2002 - Idiopathic thrombocytopenic purpura, helicobacter pylori infection, and HLA class II alleles [3] (multiple letters) [Articolo su rivista]
Veneri, D.; Gottardi, M.; Guizzardi, E.; Zanuso, C.; Krampera, M.; Franchini, M.; Emilia, G.; Luppi, M.; Morselli, M.; Longo, G.; Torelli, G.
abstract

2002 - Long-term salvage therapy with cyclosporin A in refractory idiopathic thrombocytopenic purpura. [Articolo su rivista]
Emilia, G; Morselli, M; Luppi, Mario; Longo, G; Marasca, Roberto; Gandini, G; Ferrara, L; D'Apollo, N; Potenza, Leonardo; Bertesi, M; Torelli, Giuseppe
abstract

Treatment of severe, chronic idiopathic thrombocytopenic purpura (ITP) refractory to most usual therapies Is a difficult challenge. Little information exists on the clinical use of cyclosporin A (CyA) in the treatment of ITP. This report describes long-term treatment with CyA (median, 40 months) and follow-up (median, 36.8 months) in 12 adult patients with resistant ITR CyA used in relatively low doses (2.5-3 mg/kg of body weight per day) led to a clinical Improvement In 10 patients (83.3%). Five had a complete response (41.1%),4 a complete response to maintenance therapy (33.3%), and one a partial response (8.3%). Two patients had no response. Most patients with a response (60%) had a long-term remission (mean, 28.6 months) after discontinuation of CyA. One patient had a relapse of ITP 4 years after CyA therapy was stopped. Side effects were moderate and transient, even In patients dependent on continued CyA treatment. CyA seems to represent reasonable salvage treatment in severe, potentially life-threatening, refractory ITR

2002 - Mixed warm and cold autoimmune hemolytic anemia: complete recovery after 2 courses of rituximab treatment [Articolo su rivista]
M., Morselli; Luppi, Mario; Potenza, Leonardo; L., Facchini; S., Tonelli; D., Dini; G., Leonardi; A., Donelli; Narni, Franco; Torelli, Giuseppe
abstract

No abstract available

2002 - Response: Idiopathic thrombocytopenic purpura, Helicobacter pylori infection, and the HLA system [Articolo su rivista]
Emilia, G; Luppi, Mario; Morselli, M; Longo, G; Torelli, Giuseppe
abstract

No abstract available

2002 - Severe pancytopenia and hemophagocytosis after HHV-8 primary infection in a renal transplant patient successfully treated with foscarnet [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Rasini, V; Riva, Giovanni; Re, A; Rossi, Giorgio; Setti, G; Sandrini, S; Facchetti, F; Torelli, Giuseppe
abstract

We report the occurrence of human herpesvirus (HHV)-8 primary infection in an adult male kidney recipient. Four months after transplantation, the patient developed visceral Kaposi sarcoma, and 1 month later he presented with progressive and severe peripheral cytopenia, in the presence of a normocellular or hypercellular bone marrow (BM) with hemophagocytosis. HHV-8 was the sole pathogen detected by polymerase chain reaction either in the serum or in the BM. HHV-8 latent nuclear antigen was detected in immature progenitor cells from the BM. Immunosuppressive therapy was reduced, and the patient was treated with foscarnet for 2 weeks, leading to a dramatic normalization of blood cell counts, concomitantly with the disappearance of HHV-8 viremia. At the end of antiviral therapy, the patient received chemotherapy, and Kaposi sarcoma regressed in 2 months. Severe peripheral cytopenia may be a post-transplant complication after HHV-8 infection, for which treatment with foscarnet seems appropriate.

2001 - Angiogenesis in multiple myeloma: Correlation between in vitro endothelial colony growth (CFU-En) and clinical biological features [Articolo su rivista]
Dominici, Massimo; Luppi, Mario; F., Lanza; Campioni, D.; Barozzi, Patrizia; S., Pauli; R., Milani; F., Cavazzini; M., Punturieri; R., Trovato; G., Torelli; G., Castoldi
abstract

Mouse models and studies performed on fixed bone marrow (BM) specimens obtained from patients with multiple myeloma (MM) suggest that plasma cell growth is dependent on endothelial cell (EC) proliferation within the BM microenvironment. In order to assess whether EC overgrowth in MM reflects a spontaneous in vitro angiogenesis, BM mononucleated cells from 13 untreated (UT) MM, 20 treated (11 with melphalan and nine with DAV schedule) MM, eight patients with monoclonal gammopathy of uncertain significance (MGUS) and eight controls were seeded in an unselective medium to assess EC proliferation. Furthermore, the influence of IL6 on the EC growth was investigated. Endothelial colonies (CFU-En) appeared as small clusters, formed by at least 100 slightly elongated and sometimes bi-nucleated cells expressing factor VIII, CD31 and CD105 (endoglin). The CFU-En mean number/10(6) BM mononucleated cells in untreated MM samples (2.07 s.d. +/- 1.3) was significantly higher than in normal BM (0.28 +/- 0.48), while no difference was seen between normal BM and MGUS (0.28 +/- 0.54). Interestingly, the mean number of CFU-En in the DAV group (1.88 +/- 1.6) did not differ from the UT, while it was found to be lower in the melphalan group (0.31 +/- 0.63). The addition of anti-IL6 monoclonal antibody induced a reduction of both the plasma cells in the supernatant and the CFU-En number. This study describes a rapid and feasible assay providing support for the association between EC and plasma cells further suggesting that the in vitro angiogenesis process may parallel that observed in vivo.

2001 - BCR-ABL rearrangement is not detectable in essential thrombocythemia. [Articolo su rivista]
Emilia, G.; Marasca, Roberto; Zucchini, Patrizia; Temperani, Paola; Luppi, Mario; Torelli, Giuseppe; Lanza, F.; DE ANGELIS, C.; Gandini, D.; Castoldi, G.; Vallisa, D.; Cavanna, L.; del Senno, L.
abstract

BCR-ABL rearrangement is not detectable in essential thrombocythemia

2001 - Helicobacter pylori eradication can induce platelet recovery in idiopathic thrombocytopenic purpura [Articolo su rivista]
Emilia, G; Longo, G; Luppi, Mario; Gandini, G; Morselli, M; Ferrar, L; Amarri, S; Cagossi, K; Torelli, Giuseppe
abstract

Recent reports have suggested an association between Helicobacter pylori infection and idiopathic thrombocytopenic purpura (ITP). The prevalence of H pylori infection and the effect of its eradication in a series of 30 ITP patients were investigated. H pylori infection has been documented in 13 patients (43.33%) by C-13 urea breath test and confirmed by histologic examination. Bacterium eradication with antibiotics, obtained in 12 of 13 infected patients (92.3%), led to a complete response in 4 (33.33%) and to a partial response (platelets 90 x 10(9)/L120 x 10(9)/L) in 2 (16.66%). The response was maintained for a median of 8.33 months, but 1 patient relapsed 7 months after eradication. Search for H pylori infection seems appropriate in ITP patients at diagnosis. Bacterium eradication provides a new good option for a nonimmunosuppressive treatment in some ITP patients.

2001 - Immunoglobulin gene mutations and frequent use of VH1-69 and VH4-34 segments in hepatitis C virus-positive and hepatitis C virus-negative nodal marginal zone B-cell lymphoma [Articolo su rivista]
Marasca, Roberto; Vaccari, Paola; Luppi, Mario; Zucchini, Patrizia; Castelli, Ilaria; Barozzi, Patrizia; A., Cuoghi; Torelli, Giuseppe
abstract

Nodal marginal zone B-cell lymphoma (NMZL) is actually considered as a distinct entity that must be distinguished from extra-nodal and splenic marginal zone Lymphomas. To define the cell origin and the role of antigen stimulation we determined the nucleotide sequence of the tumor-related immunoglobulin heavy chain variable genes in 10 cases of NMZL. The results were also evaluated on the basis of the presence of chronic hepatitis C virus (HCV) infection. All 10 cases harbored VH somatic mutations with a sequence homology compared to the closest germline gene, ranging from 83.33 to 98.28%. Interestingly, different VH segments were preferentially used in HCV-positive and HCV-negative patients: three of five HCV-negative NMZLs used a VH4-34 segment joined with different D and JH segments whereas three of five HCV-positive NMZLs used a VH1-69 gene joined with a D3-22 and a JH4 segment, with very strong similarities in the CDR3s among the three different cases. These data indicate: 1) NMZL is derived from B cells that have experienced the germinal center reaction; 2) the preferential usage of a VH1-69 segment in the majority of the HCV-positive NMZL cases with similar CDR3s suggests the presence of a common antigen, probably a HCV antigen epitope, involved in the B-cell selection; and 3) the use of a VH4-34 segment suggests a role of yet unknown B-cell superantigen(s) in the selection of tumor B-cell precursors in HCV-negative NMZL.

2000 - Bone marrow failure associated with human herpesvirus 8 infection after transplantation [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Schulz, Tf; Setti, G; Staskus, K; Trovato, R; Narni, Franco; Donelli, A; Maiorana, Antonino; Marasca, Roberto; Sandrini, S; Torelli, G.
abstract

BACKGROUND: Human herpesvirus 8 (HHV-8) infection has been linked to the development of Kaposi's sarcoma and to rare lymphoproliferative disorders.METHODS: We used molecular methods, serologic methods, in situ hybridization, and immunohistochemical analyses to study HHV-8 infection in association with nonmalignant illnesses in three patients after transplantation.RESULTS: Primary HHV-8 infections developed in two patients four months after each received a kidney from the same HHV-8-seropositive cadaveric donor. Seroconversion and viremia occurred coincidentally with disseminated Kaposi's sarcoma in one patient and with an acute syndrome of fever, splenomegaly, cytopenia, and marrow failure with plasmacytosis in the other patient. HHV-8 latent nuclear antigen was present in immature progenitor cells from the aplastic marrow of the latter patient. Identification of the highly variable K1 gene sequence of the HHV-8 genome in both the donor's peripheral-blood cells and the recipients' serum confirmed that transmission had occurred. HHV-8 viremia also occurred after autologous peripheral-blood stem-cell transplantation in an HHV-8-seropositive patient with non-Hodgkin's lymphoma. Reactivation of the infection was associated with the development of fever and marrow aplasia with plasmacytosis; there was no evidence of other infections. HHV-8 transcripts and latent nuclear antigen were expressed in the aplastic marrow but not in two normal marrow samples obtained before transplantation.CONCLUSIONS: Primary HHV-8 infection and reactivation of infection may be associated with nonneoplastic complications in immunosuppressed patients.

2000 - Detection of antibodies to human herpesvirus 8 in Italian children: evidence for horizontal transmission [Articolo su rivista]
D., Whitby; Luppi, Mario; C., Sabin; Barozzi, Patrizia; AR Di, Biase; Balli, Fiorella; F., Cucci; Ra, Weiss; C., Boshoff; Torelli, Giuseppe
abstract

Human herpesvirus 8 (HHV-8), also known as Kaposi´s sarcoma associated herpesvirus (KSHV), has been shown to be the causative agent for Kaposi´s sarcoma (KS) and to be more prevalent in populations or risk groups at increased risk for KS. HHV-8 infection is rare in children from the US and the UK, but has been reported in African children. In this study we examine HHV-8 infection in children from Italy, a country with an elevated prevalence of HHV-8 in adults and high socio-economic conditions.

2000 - Detection of circulating tumor by reverse transcriptase polymerase chain reaction [1] (multiple letters) [Articolo su rivista]
Anderson, P. M.; Goeminne, J. -C.; Guillaume, T.; Federico, M.; Sabbatini, R.; Morselli, M.; Depenni, R.; Cagossi, K.; Luppi, M.; Torelli, G.; Silingardi, V.
abstract

2000 - Detection of circulating tumor cells by reverse transcriptase polymerase chain reaction of maspin in patients with breast cancer undergoing conventional-dose chemotherapy [Articolo su rivista]
R., Sabbatini; Federico, Massimo; M., Morselli; Depenni, Roberta; K., Cagossi; Luppi, Mario; Torelli, Giuseppe; Silingardi, Vittorio
abstract

Purpose: To establish, in patients with breast cancer subjected to primary conventional chemotherapy and enrolled in a prospective study, the mobilizing effect of therapy on potentially neoplastic cells by means of a reverse transcriptase polymerase chain reaction (RT-PCR) assay for mRNA of maspin, a protein related to the serpin family of protease inhibitors. Patients and Methods: Peripheral-blood samples were collected from 30 patients with histologically proven breast cancer before and 4 and 8 days after conventional chemotherapy for three consecutive courses. A total of 216 samples were screened for the presence of maspin mRNA by RT-PCR. Results: Before therapy, all samples but one were negative. After chemotherapy, 11 patients (38%) had positive samples. No difference in the rate of positivity was observed between groups defined according to initial stage, type of chemotherapy, Ki-67-related proliferative activity, or CA 15.3 expression. Conclusion: Our results confirm that RT-PCR for maspin mRNA is a sensitive assay for the study of circulating potentially neoplastic mammary cells in patients with breast cancer. Moreover, our findings indicate a marked effect of conventional-dose chemotherapy on the mobilization of these cells in breast tumors, In our series of patients, this phenomenon does not seem to be associated with other known risk factors. Finally, the data suggest, without proving, an association between the presence of circulating maspin positive cells and a higher risk of disease progression, If this association could be confirmed, then the assay could have prognostic significance. However, larger confir- matory studies are necessary.

2000 - Fatal herpesvirus-6 encephalitis in a recipient of a T-cell-depleted peripheral blood stem cell transplant from a 3-loci mismatched related donor [Articolo su rivista]
Tiacci, E; Luppi, Mario; Barozzi, Patrizia; Gurdo, G; Tabilio, A; Ballanti, S; Torelli, Giuseppe; Aversa, F.
abstract

Human herpesvirus-6 (HHV-6), like all the other herpes viruses, remains latent in host cells after primary infection but can be reactivated In immuno-compromised patients causing fever, skin rash, bone marrow (BM) suppression, pneumonitis, Sinusitis and meningoencephalitis. We describe the case of a man with chronic myelogenous leukemia who developed encephalitis associated with acute graft-versus-host disease two months after a T-cell-depleted mismatched peripheral blood stem cell transplant. Magnetic resonance images of the brain revealed multiple bilateral foci of signal abnormality. HHV-6 was the only pathogen detected in cerebrospinal fluid by PCR. treatment with both ganciclovir and foscarnet was unsuccessful and the patient gradually deteriorated and died. Other cases of HHV-6 encephalitis after bone marrow transplantation are reviewed.

2000 - Gestational thrombocytopenia - Reply [Articolo su rivista]
Emilia, Giovanni; Luppi, Mario; Marasca, Roberto; Torelli, Giuseppe
abstract

Although patients with essential thrombocythaemia (ET) do not have the Philadelphia chromosome t(9;22), which defines chronic myelocytic leukaemia, the chimeric BCR-ABL transcript mRNA from this translocation has been identified in patients with clinically typical essential thrombocythaemia”.These findings have been detected, by D Blickstein and colleagues2 in 48% of 25 patients negative for the Philadelphia chromosome who had essential thrombocythaemia, investigated by two-step nested RT-PCR. Some workers have not, however, been able to confirm these data by the same technique, in 18 and 20 such patients, respectively. [3] and [4] Moreover, others have shown the absence of the chimeric transcript in 41 patients with essential thrombocythaemia studied by the fluoresence in-situ hybridisation with a BCR-ABL probe.5We investigated 112 patients who had essential thrombocythaemia with long follow-up, by RT-PCR, and detected the BCR-ABL transcript in only one patient who progressed to myeloid blastic crisis 12 years after diagnosis. Actually, whether essential-thrombocythaemia patients negative for the Philadelphia chromosome express the BCR-ABL transcript has been a matter of controversy for several years. In studies, the apparent essential thrombocythaemia carrying the Philadelphia anomaly, cytogenetically or molecularly should probably be considered as a variant of chronic myelocytic leukaemia with thrombocytosis and often long survival, with obvious clinical and therapeutical implications.

2000 - Gestational thrombocytopenia [6] (multiple letters) [Articolo su rivista]
Minakami, H.; Sato, I.; George, J.; Emilia, G.; Luppi, M.; Marasca, R.; Torelli, G.
abstract

2000 - Late-appearing PML/RAR alpha fusion transcript with coincidental t(12;13)(p13.2;q14) in acute promyelocytic leukemia lacking the t(15;17) cytogenetic anomaly [Articolo su rivista]
Temperani, Paola; Luppi, Mario; F., Giacobbi; V., Medici; M., Morselli; Barozzi, Patrizia; Marasca, Roberto; Torelli, Giuseppe; Emilia, Giovanni
abstract

The late appearance of a cytogenetic/molecular hallmark in human leukemias is a rare event. We report on a case of acute myeloid leukemia with morphology, immunophenotype and clinical features typical of promyelocytic subtype (APL), in which the specific PML/RAR alpha gene rearrangement was molecularly detected only at second relapse of disease, without cytogenetic evidence of the t(15;17). The emergence of the PML/RAR alpha gene may be therapy-related or may represent the exceptional result of a clonal evolution during progression of neoplasia. At second relapse, a novel cell clone bearing a t(12;13)(p13.2;q14) was also observed and a molecular deletion and rearrangement of a locus at 13q14, distinct from retinoblastoma (Rb1) locus, was found. In this unusual case, the PML/RAR alpha product seems to be not essential for the expression of the promyelocytic phenotype at diagnosis and, when detectable, it is not the sole genetic defect. (C) 2000 Elsevier Science Inc. All rights reserved.

2000 - Molecular evidence of organ-related transmission of Kaposi sarcoma-associated herpesvirus or human herpesvirus-8 in transplant patients [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; G., Santagostino; R., Trovato; Tf, Schulz; Marasca, Roberto; D., Bottalico; L., Bignardi; Torelli, Giuseppe
abstract

In transplant patients, Kaposi sarcoma (KS)-associated herpesvirus or human herpesvirus-8 (HHV-8) infection is associated with the development of KS, primary effusion lymphoma and Castleman disease. Whether HHV-8 is either reactivated in the recipient or transmitted by the donor has been investigated so far only by serologic studies. Thus, we addressed the issue of HHV-8 transmission in the transplantation setting by molecular methods. We exploited the high level variability of the orf-K1 gene and the polymorphism of the orf-73 gene of the HHV-8 genome to assess the genetic relatedness of the HHV-8 strains identified in the posttransplant KS lesions that developed, simultaneously, 20 months after transplantation, in 2 recipients of twin kidneys from the same cadaver donor. The 100% identity of nucleotide sequence of the most variable viral region and the presence of the same, single orf-73 type in both patients provides strong molecular evidence of organ-related transmission of HHV-8 in the setting of transplantation.

2000 - Nonmalignant disease associated with human herpesvirus 8 reactivation in patients who have undergone autologous peripheral blood stem cell transplantation [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Tf, Schulz; R., Trovato; A., Donelli; Narni, Franco; J., Sheldon; Marasca, Roberto; Torelli, Giuseppe
abstract

Fever, cutaneous rash, and hepatitis-for which an infectious cause was suspected-developed in an Italian patient with non-Hodgkin lymphoma after autologous peripheral blood stem cell (PBSC) transplantation. Polymerase chain reaction (PCR) with degenerate primers for the highly conserved DNA polymerase gene of herpesviruses detected herpesvirus sequences 100% identical to human herpesvirus-8 (HHV-8) in serial cell-free serum samples, collected immediately before or concomitant with the occurrence of clinical symptoms; no other common infections were documented. The presence of the HHV-8 genome (clade C) was confirmed by PCR with HHV-8-specific primers for orf 26 and orf-K1, HHV-8 viremia was undetectable either before transplantation or when the patient was clinically asymptomatic. Semiquantitative PCR analysis showed variations of the viral load correlating with the clinical status. Anti-HHV-8 antibodies were detected before and after transplantation by an immunofluorescence assay for lytic antigens. Active HHV-8 infection may be associated with nonmalignant illness after PBSC/bone marrow transplantation. (Blood. 2000;96:2355-2357)

2000 - PCR with degenerate primers for highly conserved DNA polymerase gene of the herpesvirus family shows neither human herpesvirus 8 nor a related variant in bone marrow stromal cells from multiple myeloma patinets. [Articolo su rivista]
Dominici, Massimo; Luppi, Mario; D., Campioni; F., Lanza; Barozzi, Patrizia; R., Milani; S., Moretti; G., Nadali; R., Spanedda; R., Trovato; Torelli, Giuseppe; G., Castoldi
abstract

The possibility has been raised that either a human herpesvirus-8 (HHV-8) variant or a novel, unidentified, gamma-herpesvirus related to HHV-8 is frequently associated with multiple myeloma (MM), which could explain the lack of antibodies to HHV-8 antigens and the discordant results from polymerase chain reaction (PCR) studies of HHV-8-specific sequences in MM patients. Thus, we used a sensitive PCR assay with degenerate primers targeting the highly conserved DNA polymerase gene of the herpesvirus family to examine the long-term cultures of bone marrow stromal cells (BMSCs) from 19 MM, 3 monoclonal gammopathies of undetermined significance and 6 control patients. Both the culture supernatant and the adherent stromal layer were examined from the 2nd until the 8th week of culture to assess the immunophenotype of the various cell types harvested for the molecular analysis. BMSCs consisted of a mixed population of fibroblast, macrophage, dendritic and endothelial cells. An amplified product of the expected size was obtained only in 3 MM cases, both in the adherent and nonadherent fractions. Direct sequencing and alignment of the nucleotide and amino acid sequences showed that the DNA sequences were 100% identical to Epstein-Barr virus (EBV) DNA. The PCR positivity was due to the presence of EBV-infected lymphoblastoid cells with plasmacytoid features, expressing the EBV-encoded latent membrane protein-1 and detectable either in the stromal cells or in the culture supernatant. Our data do not support a causal role of either HHV-8 or a novel herpesviral variant related to HHV-8 in MM.

1999 - Cellular localization of human herpesvirus 8 in nonneoplastic lymphadenopathies and chronic interstitial pneumonitis by in situ polymerase chain reaction studies [Articolo su rivista]
Trovato, R; Luppi, Mario; Barozzi, Patrizia; Da Prato, L; Maiorana, Antonino; Lico, S; Marasca, Roberto; Torricelli, Pietro; Torelli, Giuseppe; Ceccherini Nelli, L.
abstract

Objectives: To study the cellular localization of human herpesvirus 8 (HHV-8) in rare cases of HHV-8 infection from Italy that are associated neither with human immunodeficiency virus (HIV) infection nor Kaposi's sarcoma (KS). Methods: The presence and distribution of HHV-8-infected cells was investigated by direct in situ polymerase chain reaction (PCR) in the lymph node tissues from 2 patients with reactive lymphadenopathies with florid follicular hyperplasia and increased vascularity and in the lung tissue from 1 patient with chronic interstitial pneumonitis. Results: HHV-8 was localized in lymphoid and monocyte-macrophage cells scattered in the interfollicular regions of both lymph nodes but not in endothelial cells. In the lung tissue, HHV-8 was found in the inflammatory cells infiltrating the interalveolar interstitium, in endothelial cells of the pulmonary vasculature, and in rare pneumocytes. Conclusions: HHV-8 can infect nonneoplastic lymph nodes of immunocompetent subjects, and the distribution of infected cells outside of the germinal centers resembles that of Epstein-Barr virus (EBV)-infected cells in the lymph nodes in the course of infectious mononucleosis. Endothelial cells and pneumocytes may be a target of HHV-8 infection out of the KS setting, at least in the presence of a chronic inflammatory process.

1999 - Erratum: Cellular localization of human herpesvirus 8 in nonneoplastic lymphadenopathies and chronic interstitial pneumonitis by in situ polymerase chain reaction studies (Journal of Human Virology (January/February 1999) 2:1 (38-44)) [Articolo su rivista]
Trovato, R.; Luppi, M.; Barozzi, P.; Da Prato, L.; Maiorana, A.; Lico, S.; Marasca, R.; Torricelli, P.; Torelli, G.; Ceccherini-Nelli, L.
abstract

1999 - Expression of cell-homologous genes of human herpesvirus-8 in human immunodeficiency virus-negative lymphoproliferative diseases [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Maiorana, Antonino; R., Trovato; Marasca, Roberto; M., Morselli; K., Cagossi; Torelli, Giuseppe
abstract

Human herpesvirus-8 (HHV-8) genome encodes for genes homologous to human cellular genes such as interleukin-6 (IL-6), Cyclin-D, BCL-5, and IL-8 receptor (G-protein-coupled receptor [GCR]), We used reverse transcriptase-polymerase chain reaction to study the expression of these viral genes in lymphoproliferative disorders associated with HHV-8 infection, None of these genes was expressed in 1 case of benign, localized Castleman´s disease (CD), and only viral IL-6 and viral Cyclin-D were transcribed in 2 cases of benign lymphadenopathies with giant germinal center hyperplasia and increased vascularity. In contrast, all 4 genes were transcribed in 1 case of multicentric CD of plasma cell type with aggressive clinical course and in 1 primary effusion lymphoma cell line. Our study provides the evidence that various HHV-8 genes, homologous to cellular genes involved in control of proliferation and apoptosis, may be differently expressed in different lymphoid disorders in vivo.

1999 - Fine mapping of an apparently targeted latent human herpesvirus type 6 integration site in chromosome band 17p13.3 [Articolo su rivista]
Morris, C; Luppi, Mario; Mcdonald, M; Barozzi, Patrizia; Torelli, Giuseppe
abstract

An unusually high level of latent HHV-6 infection has been documented in the peripheral blood and/or bone marrow cells of a small group of patients with predominantly malignant lymphoid disorders, and in at least one healthy individual, We have shown previously in peripheral blood mononuclear cells (PBMCs) of three patients, two with a history of lymphoma and one with multiple sclerosis, a specific target site for latent integration of the full-length HHV-6 viral genome on the distal short arm of chromosome 17, in band p13.3. Fluorescence in situ hybridization (FISH) procedures were used to map more precisely the location of the viral integration site in one of those patients, relative to two known oncogenes mapped previously, namely CRK, and the more telomeric ABR oncogene. It is shown that the HHV-6 integration site is located at least 1,000 kb telomeric of ABR, and is very likely to map close to or within the telomeric sequences of 17p. This finding is significant given that human telomeric-like repeats flank the terminal ends of the HHV-6 genome. Cytogenetic studies showed evidence of karyotype instability in the peripheral blood cells infected latently. J. Med. Virol. 58:69-75, 1999.

1999 - Human herpesvirus 6 latently infects early bone marrow progenitors in vivo [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; C., Morris; Maiorana, Antonino; R., Garber; G., Bonacorsi; A., Donelli; Marasca, Roberto; A., Tabilio; Torelli, Giuseppe
abstract

We have studied the in vivo tropism of human herpesvirus 6 (HHV-6) for hemopoietic cells in patients with latent HHV-6 infection. Having used a variety of cell purification, molecular, cytogenetic, and immunocytochemical procedures, we report the first evidence that HHV-6 latently infects early bone marrow progenitor cells and that HHV-6 may be transmitted longitudinally to cells which differentiate along the committed pathways.

1999 - Human herpesvirus-8 infection in hemodialysis patients from northern Italy. [Articolo su rivista]
Luppi, Mario; Vandelli, L.; Whitby, D.; Savazzi, A. M.; Barozzi, Patrizia; Medici, G.; Albertazzi, Alberto; Torelli, Giuseppe
abstract

No abstract available

1999 - Lack of confirmation of an association between HTLV-I infection and myelodysplastic syndrome [Articolo su rivista]
M., Morselli; Luppi, Mario; Barozzi, Patrizia; Dominici, Massimo; Temperani, Paola; D., Campione; F., Lanza; R., Trovato; Marasca, Roberto; G., Longo; G., Emilia; Torelli, Giuseppe
abstract

No abstract available

1999 - Missense mutations in the PML/RAR alpha ligand binding domain in ATRA-resistant As2O3 sensitive relapsed acute promyelocytic leukemia [Articolo su rivista]
Marasca, Roberto; Zucchini, Patrizia; S., Galimberti; G., Leonardi; Vaccari, Paola; A., Donelli; Luppi, Mario; M., Petrini; Torelli, Giuseppe
abstract

Background and Objectives. Acute promyelocytic leukemia is characterized by the chromosomal translocation t(15;17) which yields the fusion product PML/RAR alpha. Art-trans retinoic acid probably induces differentiation of atypical promyelocytes and clinical remission in APL patients by binding to the ligand binding domain (LBD) of the RAR alpha portion of the PML-RAR alpha chimeric protein. Structural alterations of the LED of the PML/RAR alpha have been revealed in ATRA-resistant APL cell lines and in a few APL patients with acquired clinical resistance to ATRA therapy. Two APL relapsed patients with clinical resistance to ATRA therapy were evaluated for the presence of nucleotide mutations in the LED of PML/RAR alpha gene and then treated with arsenic trioxide (As2O3). Design and Methods. DNA fragments from the LED of the PML/RAR alpha: chimeric transcript were obtained by reverse-transcribed polymerase chain reaction. Direct sequencing was performed by an unambiguous bidirectional automatic analysis. Samples representative of APL onset and relapse were analyzed from both patients. Results. In both patients, at the ATRA-resistant relapse, a missense point mutation in the LED of the PML/RAR alpha gene was found. The mutations, absent at APL onset, led to an Arg272Gln and to an Arg276Trp amino acid substitution, according to the sequence of the RAR alpha protein. Both patients had complete clinical and hematologic remission after treatment with As2O3. Interpretation and Conclusions. LED missense mutations appear to be a significant mechanism of acquired ATRA-resistance in vivo, closely related to clinical APL relapse. The two cases reported here provide the first in vivo evidence of Apt, relapsed patients, who have become ATRA-resistant for molecular reasons, being sensitive to arsenic trioxide.

1999 - Neither Human Herpesvirus 8 nor a related variant could be detected in bone marrow stromal cells from multiple myeloma patients. [Abstract in Rivista]
Dominici, Massimo; Luppi, Mario; D., Campioni; F., Lanza; Barozzi, Patrizia; R., Milani; A., Latorraca; S., Moretti; Torelli, Giuseppe; G. L., Castoldi
abstract

The possibility has been raised that either a human herpesvirus- 8 (HHV-8) variant or a novel, unidentified, g-herpesvirus related to HHV-8 is frequently associated with multiple myeloma (MM), which could explain the lack of antibodies to HHV-8 antigens and the discordant results from polymerase chain reaction (PCR) studies of HHV-8-specific sequences in MM patients. Thus, we used a sensitive PCR assay with degenerate primers targeting the highly conserved DNA polymerase gene of the herpesvirus family to examine the longterm cultures of bone marrow stromal cells (BMSCs) from 19 MM, 3 monoclonal gammopathies of undetermined significance and 6 control patients. Both the culture supernatant and the adherent stromal layer were examined from the 2nd until the 8th week of culture to assess the immunophenotype of the various cell types harvested for the molecular analysis. BMSCs consisted of a mixed population of fibroblast, macrophage, dendritic and endothelial cells. An amplified product of the expected size was obtained only in 3 MM cases, both in the adherent and nonadherent fractions. Direct sequencing and alignment of the nucleotide and amino acid sequences showed that the DNA sequences were 100% identical to Epstein-Barr virus (EBV) DNA. The PCR positivity was due to the presence of EBV-infected lymphoblastoid cells with plasmacytoid features, expressing the EBV-encoded latent membrane protein-1 and detectable either in the stromal cells or in the culture supernatant. Our data do not support a causal role of either HHV-8 or a novel herpesviral variant related to HHV-8 in MM.

1999 - PCR con primer degenerati non evidenzia sequenze virali di HHV-8 ne di una sua variante in cellule stromali midollari di pazienti affetti da gammopatie monoclonali [Abstract in Rivista]
Dominici, Massimo; Luppi, Mario; D., Campioni; F., Lanza; Barozzi, Patrizia; R., Milani; S., Moretti; R., Spanedda; Torelli, Giuseppe; G. L., Castoldi
abstract

The association of human herpesvirus-8 (HHV-8) infection with multiple myeloma (MM) is still controversial. The possibility has been raised that either a HHV-8 variant or a novel, as yet unidentified, γ-herpesvirus related to HHV-8, are frequently associated with MM, which could explain the lack of antibodies to HHV-8 antigens and the discordant results from polymerase chain reaction studies (PCR) of HHV-8 specific genomic sequences in MM patients. Thus, we used a sensitive PCR assay with degenerate primers targeting the highly conserved DNA polymerase gene of the herpesvirus family to examine the long term-cultures (LTC) of bone marrow stromal cells (BMSCs) from 21 MM, 3 monoclonal gammopathy of undetermined significance (MGUS) and 6 control patients. Both the culture supernatant and the adherent stromal layer were examined from the 2nd until the 8th week of culture to assess the immunophenotype of the various cell types harvested for the molecular analysis. BMSCs consisted of a mixed population of fibroblast, macrophage, dendritic and endothelial cells. An amplified product of the expected size was obtained only in 3 MM cases, both in the adherent and non adherent fractions. Direct sequencing and alignment of the nucleotide and amino acidic sequences showed that the DNA sequences were 100% identical to Epstein-Barr virus DNA. PCR with specific primers targeting the latent membrane protein-1 (LMP-1) gene confirmed the presence of EBV. The PCR positivity was due to the presence of EBV infected lymphoblastoid cells (LCLs) with plasmacytoid features, expressing the LMP-1 protein and detectable either in the stromal cells or in the culture supernatant. Our data do not support a causal role of either HHV-8 or a novel herpesviral variant related to HHV-8 in MM, not even in the presence of EBV co-infection.

1999 - The oncogenic 30 and 69 bp deletion variants of the EBV LMP-1 gene are common in HIV-negative lymphoproliferations, both malignant and benign [Articolo su rivista]
Barozzi, Patrizia; Luppi, Mario; K., Cagossi; Maiorana, Antonino; Marasca, Roberto; T., Artusi; S., Poggi; S. A., Pileri; Torelli, Giuseppe
abstract

BACKGROUND: In vitro studies have shown that the 30 and 69 base pair (bp) deletion variants of the latent membrane protein (LMP)-1 gene of the Epstein-Barr virus (EBV) have a higher transforming capacity than the wild-type variant. In recent years these studies have triggered an in vivo search for such potentially oncogenic variants in lymphoid tissues. PATIENTS AND METHODS: We used polymerase chain reaction (PCR) to investigate the prevalence of LMP-1 gene variants in EBV-positive lymph nodes from 60 HIV-negative Italian patients with benign and malignant lymphoid disorders. RESULTS: The 30 bp variant was detected in 10 of 39 (25.6%) malignant lymphomas but also in 4 of 13 (30%) reactive lymphadenitis with follicular hyperplasia. Of note is the fact that the 69 bp variant was detected in three cases of malignant lymphoproliferation but also in two cases of localized Castleman's disease of hyalin vascular type. CONCLUSIONS: The molecular detection of the oncogenic variants of the LMP-1 gene in a lymph node biopsy as an indicator of the aggressiveness of the EBV-associated lymphoproliferative disease must be considered with caution. The relatively high frequency of the 69 bp variant in our series compared with that reported in the literature probably reflects a different incidence of LMP-1 variants in healthy populations from different geographical areas.

1999 - Variability and evolution of Kaposi's sarcoma-associated herpesvirus in Europe and Africa. International Collaborative Group. [Articolo su rivista]
Cook, Pm; Whitby, D; Calabro, Ml; Luppi, Mario; Kakoola, Dn; Hjalgrim, H; Ariyoshi, K; Ensoli, B; Davison, Aj; Schulz, T. F.
abstract

OBJECTIVE: To study the evolution of Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus type 8 in Europe and Africa.DESIGN AND METHODS: PCR and sequence analysis of the variable viral membrane glycoprotein gene K1 in 58 tumour and peripheral blood samples from patients with AIDS-related Kaposi's sarcoma (KS), 'classic' (HIV-negative) KS, transplant KS, Multicentric Castleman's Disease, other lymphoproliferative disorders, and healthy KSHV-infected individuals from the UK, Denmark, Sweden, Italy, Spain, Iceland, The Faroe Islands, Greece, The Gambia and Uganda.RESULTS: Three major groups of K1 sequences were found: A, B and C, as defined previously. The K1 gene has evolved, both within and between these three groups, under positive selection. KSHV group B strains predominate in Africa and are more distant from groups A and C, found in Europe, than A and C are from each other. Within group C two subgroups, C' and C", can be identified. Subgroup C" is more closely related to group A in a region of the K1 protein and appears to be phylogenetically close to the branchpoint between A and C. Group A and C strains are currently found in both HIV-1-infected and -uninfected Europeans, and were already present in Europe before the start of the AIDS epidemic. We found some examples of closely related K1 sequences in Italy and Denmark, but in general KSHV strains in Europe did not cluster geographically.CONCLUSION: KSHV strains in East and West Africa are closely related but phylogenetically distant from those in Europe. The two major KSHV groups in Europe are more closely related, with some strains adopting an intermediate phylogenetic position. In Europe, KSHV strains may have been disseminated at least several decades ago. Variability in the K1 region is driven by selection and does not correlate with different KSHV-related pathologies or geographic regions where clinically more aggressive HIV-negative KS ('endemic' KS) is more common.

1998 - Absence of a directly causative role for human herpesvirus 7 in human lymphoma and a review of human herpesvirus 6 in human malignancy [Articolo su rivista]
Berneman, Zn; Torelli, Giuseppe; Luppi, Mario; Jarrett, Rf
abstract

In search of a (new) viral etiological agent, we screened 64 lymph node samples from Hodgkin's disease (HD) and 43 samples (32 lymph node and 11 skin biopsies) from non-Hodgkin's lymphoma (NHL) for human herpesvirus 7 (HHV-7). Twenty-nine control samples were tested as well, including 17 with benign lymphadenopathy. None of the samples tested positive by Southern blot hybridization using HHV-7-specific probes, We conclude that there is no major HHV-7 load in human lymphoma and that HHV-7 is not likely to be directly involved in its etiology. This is in contrast to a small minority of human lymphoproliferative diseases in which HHV-6 can be found at high copy number, but in which an etiological role is still uncertain.

1998 - Age- and sex-specific seroprevalence of human herpesvirus 8 in Jamaica - Response [Articolo su rivista]
Luppi, Mario; Whitby, D; Barozzi, Patrizia; Boshoff, C; Weiss, R; Cucci, F; Torelli, Giuseppe
abstract

No abstract available

1998 - Age- and sex-specific seroprevalence of human herpesvirus 8 in Jamaica [3] (multiple letters) [Articolo su rivista]
Manns, A.; Strickler, H. D.; Hanchard, B.; Manassaram, D. M.; Waters, D.; Ablashi, D. V.; Luppi, M.; Whitby, D.; Barozzi, P.; Boshoff, C.; Weiss, R.; Cucci, F.; Torelli, G.
abstract

1998 - Chronic myeloid leukemia with thrombocythemic onset may be associated with different BCR/ABL variant transcripts [Articolo su rivista]
Emilia, Giovanni; Luppi, Mario; Ferrari, Mg; Temperani, Paola; Marasca, Roberto; Giacobbi, F; Vaccari, Paola; Bandieri, E; Di Donato, C; Carapezzi, C; Torelli, Giuseppe
abstract

Ph-positive chronic myeloid leukemia (CML) mimicking essential thrombocythemia (ET) at onset seems to be a distinct clinical entity. Whether this rare clinical form of CML is associated with single, specific variants of BCR/ABL transcripts is a matter of debate. Among 82 consecutive patients with Ph-positive CML, we identified 3 patients in which the disease mimicked ET at presentation, because-of marked thrombocytosis and moderate leukocytosis, with few immature myeloid cells in peripheral blood and blood basophilia in 2 of them. Molecular analysis with the reverse transcriptase-polymerase chain reaction technique showed the presence of b2a2, b3a2, and b3a2-b2a2 transcript variants in the three patients, respectively. The results of our study together with a review of literature data suggest that different BCR/ABL transcript variants may occur in CML mimicking ET, without an apparently significant prevalence of one type.

1998 - Clinico-pathological characterization of hepatitis C virus-related B-cell non-Hodgkin's lymphomas without symptomatic cryoglobulinemia [Articolo su rivista]
Luppi, Mario; G., Longo; Mg, Ferrari; Barozzi, Patrizia; Marasca, Roberto; M., Morselli; C., Valenti; Mascia, Maria Teresa; L., Vandelli; D., Vallisa; L., Cavanna; Torelli, Giuseppe
abstract

Background. Epidemiological evidence has suggested an association between hepatitis C virus (HCV) infection and B-cell lymphoproliferation. We studied the prevalence of HCV infection in a series of de novo B-cell non-Hodgkin's lymphoma (B-NHL) cases and correlated virological findings with clinico-histological features. Patients and methods. One hundred fifty-seven patients with de novo B-NHL were included in the study. Their serum was examined by ELISA and RIBA for the presence of anti-HCV antibodies, and either the peripheral blood mononuclear cells or the pathology tissues of all of the patients were examined by reverse transcriptase polymerase chain reaction for the presence of HCV RNA sequences, Results: HCV infection occurred in 22.3% of B-NHL patients and was documented before the diagnosis in about halfof the positive cases. Of interest. HCV infection was more frequently found in follicular center; marginal zone and diffuse large-cell lymphoma types, but was not associated with symptomatic cryoglobulinemia. The median survival time was 48 months in HCV-positive and 52 months in HCV-negative B-NHL patients. Conclusions. Our findings strengthen the pathogenetic link between HCV and B-NHL and show that HCV infection may be associated with the malignant proliferation of defined B-cell subsets other than the immunoglobulin Mk B-cell subset involved in the pathogenesis of mixed cryoglobulinemia type II and associated lymphoplasmacytoid lymphoma type. HCV-related liver disease did not affect the survival of our B-NHL patients.

1998 - Expression of human herpesvirus-6 antigens in benign and malignant lymphoproliferative diseases [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; R., Garber; Maiorana, Antonino; G., Bonacorsi; T., Artusi; R., Trovato; Marasca, Roberto; Torelli, Giuseppe
abstract

Immunohistochemistry was used to look for the expression of human herpesvirus-6 (HHV-6) antigens in a well characterized series of benign, atypical, and malignant lymphoid lesions, which tested positive for the presence of HHV-6 DNA, A panel of specific antibodies against HHV-6 antigens, characteristic either of the early (p41) or late (p101K, gp106, and gp116) phases of the viral cycle, was applied to the lymphoid tissues from 15 non-Hodgkin's lymphomas, 14 Hodgkin's disease cases, 5 angioimmunoblastic lymphadenopathies with dysproteinemia, 14 Reactive lymphadenopathies, and 2 cases of sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease). In lymphomatous tissues, the expression of late antigens was documented only in reactive cells, and mainly in plasma cells. Of interest, the expression of the early p41 antigen was detected in the so-called mummified Reed-Sternberg cells, in two Hodgkin's disease cases. In reactive lymphadenopathies, the HHV-6 late antigen-expressing cells were plasma cells, histiocytes, and rare granulocytes distributed in interfollicular areas. In both cases of Rosai-Dorfman disease, the p101K showed an intense staining in follicular dendritic cells of germinal centers, whereas the gp106 exhibited an intense cytoplasmic reaction in the abnormal histiocytes, which represent the histological hallmark of the disease. The expression of HHV-6 antigens is tightly controlled in lymphoid tissues. The lack of HHV-6 antigen expression in neoplastic cells and the limited expression in degenerating Reed-Sternberg cells argue against a major pathogenetic role of the vines in human lymphomagenesis, The detection of a rather unique pattern of viral late antigen expression in Rosai-Dorfman disease suggests a possible pathogenetic involvement of HHV-6 in some cases of this rare lymphoproliferative disorder.

1998 - Hepatitis C virus genotype distribution in B-Cell non-Hodgkin lymphoma [Articolo su rivista]
Luppi, Mario; Ferrari, Mg; Torelli, Giuseppe
abstract

No abstract available

1998 - Human herpesvirus 8 seroprevalence in blood donors and lymphoma patients from different regions of Italy [Articolo su rivista]
D., Whitby; Luppi, Mario; Barozzi, Patrizia; C., Boshoff; Ra, Weiss; Torelli, Giuseppe
abstract

No abstract available

1998 - Integration of human herpesvirus 6 genome in human chromosomes [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Morris, Cm; Merelli, E; Torelli, Giuseppe
abstract

No abstract available

1998 - Might essential thrombocythemia carry Ph anomaly? [Articolo su rivista]
Marasca, Roberto; Luppi, Mario; Zucchini, Patrizia; Longo, G; Torelli, Giuseppe; Emilia, G.
abstract

Might essential thrombocythemia carry Ph anomaly?

1998 - Occurrence of a novel t(11;19)(q13;q13.3) in complete remission of acute promyelocytic leukemia [Articolo su rivista]
Temperani, Paola; Luppi, Mario; F., Giacobbi; P., Vaccari; G., Longo; A., Donelli; Narni, Franco; Torelli, Giuseppe; G., Emilia
abstract

A woman with t(15;17) and PML/RAR alpha positive acute promyelocytic leukemia (APL-M3v) achieved a complete remission (CR) with cytogenetic and molecular conversion, after one-month ATRA plus idarubicin treatment. During CR, less than one-month after consolidation therapy with topoisomerase II inhibitors, a novel t(11;19) (q13;q13.3) was detected in peripheral blood stem cells and later in harvest bone marrow cells. Persisting CR and the negativity for BCL1 and PRAD1 genes rearrangement, the autotransplantation was performed, with good outcome. The patient is still in CR eighteen months post-transplant, in spite of the persistence of a small t(11;19) clone in BM cells. The emergence of a novel chromosomal change during CR of acute leukemia is a rare phenomenon. This is the first t(11;19)(q13;q13.3) described in APL. This finding raises the issue of whether the abnormal karyotypes at remission might represent a risk of tumor recurrence. The meaning of this genomic instability is yet unknown.

1998 - Polymerase chain reaction detection of human herpesvirus 8 sequences in primary central nervous system lymphomas [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Marasca, Roberto; Savarino, M; Torelli, Giuseppe
abstract

No abstract available

1998 - Still's disease, severe thrombocytopenia, and acute hepatitis associated with acute parvovirus B19 infection [Articolo su rivista]
Longo, G; Luppi, Mario; Bertesi, M; Ferrara, L; Torelli, Giuseppe; Emilia, G.
abstract

No abstract available

1997 - Detection of human herpes virus type 8 DNA sequences as a valuable aid in the differential diagnosis of Kaposi's sarcoma [Articolo su rivista]
Maiorana, Antonino; Luppi, Mario; P., Barozzi; G., Collina; Fano, Rita Adriana; Torelli, Giuseppe
abstract

The occurrence of human herpes virus type 8 (HHV-8) DNA sequences was evaluated by polymerase chain reaction in 36 cases of cutaneous Kaposi's sarcoma (KS) (classic and associated with the acquired immunodeficiency syndrome) and in 88 pathologic entities that can histologically mimic KS, such as reactive and neoplastic vascular processes and lesions featuring focal or extensive spindle cell pattern. A positive reaction was detected in 16 (70%) of 23 samples of classic KS and 10 (77%) of 13 KS cases related to the acquired immunodeficiency syndrome. In particular, 4 (50%) of 8 samples in the patch stage were found to be positive, whereas in the plaque and nodular stages, HHV-8 DNA sequences were observed in 8 (67%) of 12 and 14 (87%) of 16 cases, respectively. KS cases that had yielded previous negative results were found to be positive for HHV-8 on nested polymerase chain reaction, except two cases of classic KS in the patch stage (6% of the total number of KS, 25% of cases in the patch stage). Reactive and neoplastic vascular processes and cutaneous lesions with a spindle cell component were consistently negative, HHV-8 detection by polymerase chain reaction can represent a valuable method for diagnosing KS, particularly in small skin biopsy samples that might show histologic overlap with non-KS lesions, In early patch stage lesions, however, the diagnostic value of the method is hampered by the occurrence of cases in which HHV-8 sequences are still undetectable.

1997 - Hepatitis C virus-induced leuko-thrombocytopenia and haemolysis [Articolo su rivista]
Emilia, G; Luppi, Mario; Ferrari, Mg; Barozzi, Patrizia; Marasca, Roberto; Torelli, Giuseppe
abstract

Hepatitis C virus (HCV) has been recognized as the cause of thrombocytopenia occurring in patients with chronic hepatitis C, possibly through autoimmune mechanisms. A patient is described with B cell chronic lymphocytic leukaemia, presenting with a marked leuko-thrombocytopenia and an associated mild haemolysis secondary to HCV infection, in the absence of clinical and biochemical signs of hepatitis. Anti-HCV antibodies were detected in the serum both by ELISA and RIBA but not 2 months before the onset of cytopenia. The presence of HCV RNA was documented both in the peripheral blood mononuclear cells and in the bone marrow by reverse transcriptase polymerase chain reaction of the 5´ untranslated region of the viral genome. Of interest, HCV RNA was also found in the serum, showing that viraemia was associated with the presence of circulating anti-HCV antibodies. HCV genotyping, performed by PCR amplification of the core region, revealed the presence of an unclassifiable genotype. The hypothetical mechanisms leading to HCV-induced cytopenia in this patient are briefly discussed. Treatment with corticosteroids was effective in controlling blood cell counts, without increasing viraemia and deterioration of liver disease. HCV infection should be considered in the differential diagnosis of possible causes of cytopenia, mainly in immunosuppressed patients, even in absence of clinical and biochemical signs of hepatitis.

1997 - Human Herpesvirus 8 strain variability in clinical conditions other than Kaposi’s sarcoma [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Marasca, Roberto; Ferrari, Mg; Torelli, Giuseppe
abstract

No abstract available

1997 - Human herpesvirus 6 (HHV-6) ORF-1 transactivating gene exhibits malignant transforming activity and its protein binds to p53 [Articolo su rivista]
F., Kashanchi; J., Araujo; J., Doniger; S., Muralidhar; R., Hoch; S., Khleif; E., Mendelson; J., Thompson; N., Azumi; Jn, Brady; Luppi, Mario; Torelli, Giuseppe; Lj, Rosenthal
abstract

The 357 amino acid open reading frame 1 (OFF-1), also designated DR7, within the SalI-L fragment of human herpesvirus 6 (HHV-6) exhibited transactivation of the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) promoter and increased HIV-1 replication (Kashanchi et al., Virology, 201, 95-106, 1994). Tn the current study, the SalI-L transforming region was localized to the SalI-L-SH subfragment, Several ORFs identified in SalI-L-SH by sequence analysis were cloned into a selectable mammalian expression vector, pBK-CMV, Only pBK/ORF1 transformed NIH3T3 cells. Furthermore, cells expressing ORF-1 protein produced fibrosarcomas when injected into nude mice, whereas control cells, expressing either no ORF-1 protein or C-terminal truncated (after residue 172) ORF-1 protein, were not tumorigenic, Western blot analysis of proteins extracted from the tumors revealed ORF-1 protein. Additional studies indicated that ORF-1 was expressed in HHV-6-infected human T-cells by 18 h. Co-immunoprecipitation experiments showed that ORF-1 protein bound to tumor suppressor protein p53, and the OFF-1 binding domain on p53 was located between residues 28 and 187 of p53, overlapping with the specific DNA binding domain. Functional studies showed that p53-activated transcription was inhibited in ORF-1, but not in truncated OFF-1, expressing cells. Importantly, the truncated ORF-1 mutant also failed to cause transformation. Analysis of several human tumors by PCR revealed OFF-1 DNA sequences in some angioimmunoblastic lymphadenopathies, Hodgkin's and non-Hodgkin's lymphomas and glioblastomas. The detection of OFF-1 sequences in human tumors, while not proof per se, is a prerequisite for establishing its role in tumor development. Taken together, the results demonstrate that OFF-1 is an HHV-6 oncogene that binds to and affects p53. The identification of both transforming and transactivating activities within ORF-1 is a characteristic of other viral oncogenes and is the first reported for HHV-6.

1997 - Human herpesvirus-6 reactivation in a longitudinal study of two HIV-1 infected patients [Articolo su rivista]
Iuliano, R; Trovato, R; Lico, S; Luppi, Mario; Forastieri, G; Barsanti, La; Pizzigallo, Am; Mecocci, L; Barozzi, Patrizia; Torelli, Giuseppe; Mazzotta, F; Ceccherininelli, L.
abstract

After primary infection, human herpesvirus-6 (HHV-6) persists in latent form and can be reactivated in immunocompromised subjects. A longitudinal study of HHV-6 infection was carried out in two HIV-1 seropositive patients to provide in vivo evidence of HHV-6 reactivation. Concomitant with a significant rise of anti-HHV-6 IgG detected by IFA, a transient increase of HHV-6 viral load was shown in PBLs by PCR. During HHV-6 reactivation it was also identified either cell-free HHV-6 by PCR in plasma or IgM antibody titers. HHV-6 reactivation was followed by a temporary decrease in CD4+ count and by a progressive dramatic loss of CD4+ during the following 18 months. HHV-6 strain characterization by PCR demonstrated that first patient (MM) initially showed the B variant, followed by reactivation and persistence of the A variant, while in the second (SG) only the A variant was detected. The evidence of HHV-6 reactivation suggests its involvement in immunologic damage underlying the disease.

1997 - Kaposi's sarcoma-associated herpesvirus infection and multiple myeloma [Articolo su rivista]
Whitby, D; Boshoff, C; LUPPI, Mario; TORELLI, Giuseppe
abstract

No abstract available

1997 - Prevalence of HCV infection and second neoplasms in marginal zone lymphomas [Articolo su rivista]
Luppi, Mario; Longo, G; Ferrari, Mg; Ferrara, L; Marasca, Roberto; Barozzi, Patrizia; Morselli, M; Emilia, G; Torelli, Giuseppe
abstract

No abstract available

1997 - Relationship between BCR/ABL fusion proteins and leukemia phenotype [Articolo su rivista]
Emilia, G; Luppi, Mario; Marasca, Roberto; Torelli, Giuseppe
abstract

No abstract available

1997 - Sideroblastic anemia terminating in chronic myeloid leukemia [Articolo su rivista]
Bandieri, E; Didonato, C; Artioli, F; Carapezzi, C; Luppi, Mario; Artusi, T; Torelli, Giuseppe
abstract

No abstract available

1996 - Acquired factor VIII inhibitor at the onset of prolymphocytic leukemia. [Articolo su rivista]
Longo, G; Luppi, Mario; Ferrara, L; Torelli, U; Barbieri, U; Torelli, G.
abstract

No abstract available

1996 - Additional neoplasms and HCV infection in low-grade lymphoma of MALT type [Articolo su rivista]
Luppi, Mario; Longo, G; Ferrari, Mg; Ferrara, L; Marasca, Roberto; Barozzi, Patrizia; Morselli, M; Emilia, G; Torelli, Giuseppe
abstract

Several chronic inflammatory conditions and genetic alterations are likely to be involved in the pathogenesis of low-grade lymphoma of MALT type. In a well-characterized series of 27 patients with low-grade lymphoma of MALT type, we studied: (1) the incidence of other neoplasms, which might be indicative of genetic instability, apparently a characteristic of this disease; (2) the prevalence of serologic and molecular markers of HCV infection, which has been found in association with other lymphoproliferative disorders. Three patients had one or more additional cancers; a total of eight tumours, five of which occurred in the same patient, suggests the presence of some genetic instability in at least some cases of the disease. Rather unexpectedly, anti-HCV antibodies and HCV RNA sequences were documented in 50% of the patients examined, without elevation of serum transaminases. Of interest, the two patients with parotid and conjunctival MALT lymphomas, respectively, with a previous history of Sjogren's syndrome, were HCV positive. We suggest, for the first time, that HCV may be considered, in addition to Helicobacter pylori, as another potential infectious co-factor in the multistep pathogenesis of low-grade lymphomas of MALT type.

1996 - Expression of A-myb, but not c-myb and B-myb, is restricted to Burkitt's lymphoma, sIg(+) B-acute lymphoblastic leukemia, and a subset of chronic lymphocytic leukemias [Articolo su rivista]
Golay, J; Luppi, Mario; Songia, S; Palvarini, C; Lombardi, L; Aiello, A; Delia, D; Lam, K; Crawford, Dh; Biondi, A; Barbui, T; Rambaldi, A; Introna, M.
abstract

The A-myb gene encodes a transcription factor that is related both functionally and structurally to the v-myb oncogene. Following our observations that A-myb is expressed in a restricted subset of normal mature human B lymphocytes, with the phenotype CD38(+), CD39(-), slgM(-), we have now investigated the pattern of A-myb expression in neoplastic B cells representing the whole spectrum of B-cell differentiation and compared it to that of c-myb and B-myb. In a panel of 32 B-cell lines, A-myb was very strongly expressed in most Burkitt´s lymphoma (BL) cell lines, but weak or negative in 2 pre-B acute lymphoblastic leukemia (ALL), 4 non-Hodgkin´s lymphoma (NHL), 6 Epstein-Barr virus-immortalized lymphoblastoid cell lines, and 6 myeloma lines. Protein expression paralleled that of the RNA. We have also investigated A-myb expression in 49 fresh cases of B leukemias. Among 24 ALL, 6 were of the null and 11 of the common type and all these were negative for A-myb expression; on the other hand, all 7 B-ALL cases (slg(+)), as well as one fresh BL case with bone marrow infiltration, expressed A-myb. A-myb was undetectable in 4 prolymphocytic leukemias (PLL) but was strongly expressed in 5/20 (25%) of chronic lymphocytic leukemia (CLL) samples, In the latter A-myb did not correlate with phenotype or clinical stage. Finally, we have studied the progression of one case of CLL into Richter´s syndrome and have found that the Richter´s cells expressed about 25-fold less A-myb RNA than the CLL cells from the same patient. The pattern of c-myb and B-myb was clearly distinct from that of A-myb. C-myb and B-myb were expressed in all neoplastic groups, except in CLL cells. Thus, A-myb expression, unlike that of c-myb and B-myb, is restricted to a subset of B-cell neoplasias (in particular BL and slg(+)B-ALL) representative of a specific stage of B-cell differentiation. This expression may in part reflect expression of A-myb by the normal germinal center B cells that are the normal counterpart of these transformed B cells, The data presented strongly support a role for this transcription factor in B-cell differentiation and perhaps in B-cell transformation in some neoplasias.

1996 - Frequency and distribution of herpesvirus-like DNA sequences (KSHV) in different stages of classic Kaposi's sarcoma and in normal tissues from Italian population. [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Maiorana, Antonino; G., Collina; M. G., Ferrari; Marasca, Roberto; M., Morselli; E., Rossi; L., Ceccherini Nelli; Torelli, Giuseppe
abstract

The frequency and distribution of herpesvirus-like DNA sequences (KSHV) were investigated by PCR in the pathologic skin lesions of a series of 22 HIV-negative elderly patients with classic Kaposi´s sarcoma (KS) from Italy, one of the few regions of the world where classic KS is prevalent. Viral sequences were clearly identifiable in 15 cases, in particular in 2 of 5 patch, in 3 of 6 plaque and in 10 of 11 nodular lesions. Our findings confirm the association of these herpesvirus-like DNA sequences with KS in unrelated populations, providing evidence of the putative KS-associated agent in all different histologic lesions of the disease, mainly in the nodular stage. The search for other herpesviruses by PCR showed that Epstein-Barr virus (EBV) sequences were present in 7 of 22 pathologic skin lesions. In 4 cases, both EBV and KSHV were present. On the contrary, all 22 classic KS specimens were negative for human herpesvirus-6 sequences. Two of 3 patch and the 1 nodular lesions from AIDS-related KS patients examined were positive for KSHV but negative for both EBV and HHV-6 sequences. Furthermore, we evaluated the prevalence of KSHV sequences in the normal population of the same geographical area. Thirteen peripheral blood mononuclear cell samples, 9 salivary gland tissues and 6 saliva samples from healthy subjects were invariably found negative for KSHV, using the same PCR technique. Of interest, 2 of 11 hyperplastic tonsils harboured these herpesvirus-like sequences, suggesting that, like other herpesviruses, the KS-associated agent may be harboured in a proportion of normal individuals and tonsils may represent at least one of the possible reservoirs of this putative lymphotropic gamma-herpesvirus in vivo.

1996 - HHV-8-associated primary cerebral B-cell lymphoma in HIV-negative patient after long-term steroids [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Marasca, Roberto; Savarino, M; Torelli, Giuseppe
abstract

No abstract available

1996 - Hepatitis C virus infection in subsets of neoplastic lymphoproliferations not associated with cryoglobulinemia [Articolo su rivista]
Luppi, Mario; Ferrari, Mg; Bonaccorsi, G; Longo, G; Narni, Franco; Barozzi, Patrizia; Marasca, Roberto; Mussini, C; Torelli, Giuseppe
abstract

Hepatitis C virus (HCV) is both hepatotropic and lymphotropic and a dear-cut association has been proposed between HCV infection and mixed cryoglobulinemia (MC), a benign lymphoproliferative disorder, which sometimes evolves into a frank malignant B cell non-Hodgkin's lymphoma (B-NHL). Moreover, the presence of antibodies to HCV, as well as of HCV-specific genomes has been reported in the sera of over 37% patients with B-NHL, not associated with MC. Thus, we decided to perform both a serologic and a molecular study to give insights into a possible relationship between HCV infection and neoplastic lymphoproliferations. We used ELISA and RIBA tests to show that anti-HCV antibodies were present in the serum of 29 out of 69 unselected B-NHL patients (42%), while seropositivity in a healthy population was about 1%. The prevalence of anti-HCV antibodies was low in definite subsets of B lymphoid disorders, including multiple myeloma, Waldenstrom's macroglobulinemia and monoclonal gammopathies of undetermined significance. Then, using reverse transcriptase polymerase chain reaction, we detected HCV sequences directly in the pathologic lymph node biopsies in 13 out of 34 B-NHL cases, and in particular in six out of eight low-grade lymphomas of MALT type and in five out of eight centroblastic-centrocytic follicular lymphomas. In contrast, the peripheral blood samples from 10 B cell chronic lymphocytic leukemia patients resulted negative for the presence of HCV genomes. Similarly, viral sequences were absent in 10 T cell NHL, while only one out of the 14 Hodgkin's disease cases tested resulted positive. Finally, we used a PCR-based assay to characterize the genotypes (I-IV) present in the positive lymphomatous tissues. The presence of both serologic and molecular markers of HCV infection in a high percentage of certain types of B-NHL, not associated with cryoglobulinemia, and its absence from other lymphoproliferative diseases extends the spectrum of HCV-associated lymphoproliferations arguing in favor of some role of this viral infection in the pathogenesis of the malignant proliferation of definite B lymphoid populations.

1996 - Human herpesvirus 8 and interstitial pneumonitis in an HIV-negative patient [Articolo su rivista]
Luppi, Mario; Torelli, Giuseppe
abstract

No abstract available

1996 - Human herpesvirus-8 DNA sequences in Human Immunodeficiency Virus-negative angioimmunoblastic lymphadenopathy and benign lymphadenopathy with giant germinal center hyperplasia and increased vascularity [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Maiorana, Antonino; T., Artusi; R., Trovato; Marasca, Roberto; M., Savarino; L., Ceccherini Nelli; Torelli, Giuseppe
abstract

Human herpesvirus-8 (HHV-8) DNA sequences have been reported to be strictly associated not only with various forms of Kaposi´s sarcoma but also with an unusual subgroup of acquired immunodeficiency syndrome (AIDS)-related B-cell lymphomas. A possible relation of this putative virus also with multicentric Castleman´s disease (MCD) has been recently suggested. We used polymerase chain reaction to look for the presence of HHV-8 sequences in a well characterized series of benign, atypical, and malignant lymphoid tissues from 45 Hodgkin´s disease and 43 non-Hodgkin´s lymphoma (NHL) cases, as well as from 5 MCD, 15 angioimmunoblastic lymphadenopathy (AILD), and 23 benign lymphadenopathy cases. Among the 38 AIDS-related lymphoid lesions, only 1 NHL and 1 persistent generalized lymphadenopathy (PGL) case were positive. Furthermore, among the 92 non-AIDS-related lymphoproliferative disorders, HHV-8 sequences were detected in 3 classic AILD cases and in 4 reactive lymphadenopathies. Six of 9 HHV-8 positive lymphoid lesions (1 NHL, 1 PGL, 1 AILD, and 3 reactive lymphadenopathy cases) were also positive for Epstein-Barr viral sequences. The four human immunodeficiency virus (HIV) negative lymphadenopathies positive for HHV-8 sequences showed an almost identical histology, characterized by a predominantly follicular lesion, with giant germinal center hyperplasia, and increased vascularity, resembling HIV-related lymphadenopathy and MCD. Our results, while providing the first evidence of the presence of HHV-8 sequences in AILD cases, suggest a possible association of these herpesviral sequences with a distinct hystologic type of non-neoplastic lymphadenopathy, not associated with other common herpes infections.

1996 - Lymphotropic herpes virus (EBV, HHV-6, HHV-8) DNA sequences in HIV negative Castleman's disease [Articolo su rivista]
Barozzi, Patrizia; Luppi, Mario; Masini, L; Marasca, Roberto; Savarino, M; Morselli, M; Ferrari, Mg; Bevini, M; Bonacorsi, G; Torelli, Giuseppe
abstract

Aim-To evaluate the possible involvement of lymphotropic herpes viruses in Castleman's disease. Methods-Archival formalin fixed, paraffin wax embedded biopsy specimens from 16 HIV negative patients (11 with localised and five of multicentric disease) were studied. Epstein-Barr virus (EBV), human herpes virus-6 (HHV-6) and human herpes virus-8 (HHV-8) DNA was detected using PCR. PCR was also used to characterise the EBV genomes and the clonal status of the lesions. Results-EBV sequences were identified in nine (56%) cases. The main EBV genotype detected was type 1. Two (12%) cases were positive for both HHV-6 and EBV sequences. HHV-8 sequences were detected in one case of localised Castleman's disease, the sequence of which differed from that of the HHV-8 prototype. No clonal immunoglobulin gene rearrangements were found. Conclusions-EBV DNA was detected in a substantial proportion of cases, suggesting that it may have a role in the pathogenesis of Castleman's disease, unlike HHV-6 which was detected rarely. This is the first report of HHV-8 specific sequences in the localised from of the disease.

1996 - P53 gene mutations in chronic myelogenous leukemia medullary and extramedullary blast crisis [Articolo su rivista]
Marasca, Roberto; Luppi, Mario; Barozzi, Patrizia; Ferrari, Mg; Morselli, M; Torelli, Giuseppe
abstract

Molecular alterations of the P53 gene were investigated in 27 unselected patients with chronic myelogenous leukemia (CML) blast crisis. A rearrangement of the P53 gene was evident by Southern blotting in 3 cases, one of which also showed the same alteration in the chronic phase. Single strand conformation polymorphism and sequencing analysis showed point mutations in 4 blast crisis cases. Of interest, P53 point mutations were evident in all the 3 cases of extramedullary blast crisis examined and the same point mutation was found in the myeloblastoma tissues and in the subsequent peripheral blast cells. These data indicate that: a) P53 gene mutations occur in a significant but not a large number of CML acute phase cases; b) P53 gene point mutations seem to correlate strongly with the infrequent extramedullary presentation of the blast crisis; c) the presence of the same P53 gene point mutation in extramedullary and bone marrow blast cells confirms the common clonal origin of the two cellular populations.

1996 - Reconstructive subtotal laryngectomy in the treatment of laryngeal cancer [Articolo su rivista]
Luppi, Mario
abstract

Surgical techniques of sub-total reconstructive laryngectomies can often prevent the serious impairment of total laryngectomy without having to relinquish oncological radicality. The aim of the present work has been to report on the experience in this field accrued in the ENT Department of the University of Modena from 1987 to 1992. During this period 54 subtotal laryngectomies were performed. Of these, 13 were crico-hyoido-epiglotto-pexies (C.H.E.P.) and the remaining 41 were crico-hyoido-pexies (C.H.P.). The criteria suggested in the literature was adopted for tumor evaluation, surgical indications and contraindications. All the patients had a follow-up of at least 2 years and 31 of them have had at least 5 years of follow-up. There were 9 deaths: 3 due to intervening illnesses, 2 from second primary tumors and 4 from tumor and/or node recurrences. The overall survival was 83.3% at 2 years and 77.6% at 5 years. Determinate survival (ruling out those who had died because of intervening illnesses) were 88.2% and 80%, respectively. There were 11 neoplastic repetitions of which 2 were of the primary tumor, 2 of the primary tumor plus cervical metastases, and 7 of cervical metastases alone. Recovery surgery was performed in 9 patients, 5 of whom are still alive and disease free. Functional recovery (respiration, deglutition) took place slightly earlier in C.H.E.P. than in C.H.P. but in both cases this could be shortened, particularly by introducing a rehabilitative protocol during the immediate post-operative period. In no case did it prove necessary to perform a total laryngectomy to avoid "ab ingestis" problems and only one patient has a permanent tracheostomy.

1996 - Sensitive detection of circulating breast cancer cells by reverse-transcriptase polymerase chain reaction of maspin gene [Articolo su rivista]
Luppi, Mario; Morselli, M; Bandieri, E; Federico, Massimo; Marasca, Roberto; Barozzi, P; Ferrari, Mg; Savarino, M; Frassoldati, A; Torelli, Giuseppe
abstract

Background: Maspin, a recently identified protein related to the family of serpins, is believed to play a role in human breast cancer. In an effort to improve the present methods of detection, we have developed a reverse-transcriptase polymerase chain reaction (RT-PCR) assay for maspin transcript to identify small numbers of mammary carcinoma cells in the peripheral blood and bone marrow of patients with breast cancer. Patients and methods: Five non-neoplastic mammary tissue samples, 13 breast cancer specimens as well as 17 peripheral blood and 4 bone marrow samples from normal subjects were screened for the presence of maspin mRNA by RT-PCR. The same assay was applied to peripheral blood or bone marrow samples obtained from 29 patients with stages I to IV breast cancer. Results: By RT-PCR it was possible to amplify maspin mRNA in all of the primary and metastatic breast cancer specimens, but in none of the normal hemopoietic samples from healthy donors. Thus, detection of maspin transcript in the peripheral blood or marrow of a patient known to have breast cancer is indicative of the presence of mammary carcinoma cells. In reconstitution experiments, maspin RT-PCR reliably detected 10 mammary carcinoma cells in 1 million normal peripheral-blood mononuclear cells (PBMCs). None of the 9 patients with stages I, II, or III breast cancer had maspin transcript in peripheral blood. Of note, 3 of 9 patients with stage TV breast cancer receiving systemic therapy at the time of sample collection, but only I of 11 patients with stage IV not receiving therapy, had detectable maspin transcript in peripheral blood. Moreover, 3 marrow specimens from stage TV patients tested positive by this assay. Conclusions: This pilot study suggests that maspin RT-PCR assay is a sensitive, specific and sufficiently rapid method for detection of small numbers of circulating cells and marrow micrometastases in breast cancer patients. The possibility of applying this assay in the detection of tumor cell contamination of both marrow and stem-cell apheresis harvests of breast cancer patients merits further investigation.

1996 - The new lymphotropic herpesviruses (HHV-6, HHV-7, HHV-8) and hepatitis C virus (HCV) in human lymphoproliferative diseases: An overview [Articolo su rivista]
Luppi, Mario; Torelli, Giuseppe
abstract

Considerable evidence has been accumulating in favor of a possible involvement of viral agents in the pathogenesis of human lymphomas. The most recent proposal for a lymphoma classification, the Revised European-American Classification, emphasized for the first time the pathogenetic importance of two viruses, namely Epstein-Barr virus (EBV) and human T lymphotropic virus I (HTLV-I) in the development of certain lymphoid neoplasias. However, in the last ten years new viral agents possibly related to lymphoproliferative activity have been discovered: three herpesviruses [human herpesvirus-6 (HHV-6), -7 (HHV-7) and -8 (HHV-8)] and a flavivirus, HCV. HHV-6 was isolated from the peripheral blood of patients with lymphomas and a possible role for this beta-herpesvirus in Hodgkin's disease and in angioimmunoblastic lymphadenopathy (AILD) has emerged from serological and molecular studies. HHV-7, a beta-herpesvirus genetically close to HHV-6, has not yet been found in a human disease but it utilizes CD4 as a receptor on the lymphocyte surface. Only partial HHV-8 genomic sequences have been identified so far, suggesting a genetic homology with members of the gamma-herpesvirus family, including EBV. HHV-8 sequences have been identified for the first time in all forms of Kaposi's sarcoma as well as in a variety of lymphoid disorders, including body-cavity-based non Hodgkin's lymphomas, Castleman's disease, AILD and a type of HIV-negative reactive lymphadenopathy with peculiar histologic features. Finally, after its identification as the major cause of post-transfusion and sporadic non-A, non-B hepatitis, HCV has revealed a lymphotropism both in vitro and in vivo. A strong association between HCV infection and a benign lymphoproliferative disease, essential mixed cryoglobulinemia type II, has clearly emerged both from serological and molecular studies. A possible role for this viral infection in B-cell non Hodgkin's lymphomas not associated with cryoglobulinemia has also been proposed recently. The present work offers an overview of the huge amount of experimental and clinical observations supporting the possible involvement of these new lymphotropic viruses in human lymphoproliferative diseases.

1995 - Daily variation of immunoreactive serum interleukin-6 levels in multiple myeloma [Articolo su rivista]
Bandieri, E.; Luppi, Mario; Luppi, G.; Federico, Massimo; Sabbatini, R.; Torelli, Giuseppe; Piccinini, Lino
abstract

NO ABSTRACT

1995 - Frequency of human herpesvirus type 6 (HHV-6) genome detection in AIDS-related lymphoproliferative disorders [Articolo su rivista]
Trovato, R; Luppi, Mario; Vago, L; Torelli, G; Moroni, M; Ceccherini Nelli, L.
abstract

No abstract available

1995 - Geographic sequence variation of latent membrane protein 1 gene of Epstein-Barr virus in Hodgkin's lymphomas. [Articolo su rivista]
Lin, Jc; Lin, Sc; Luppi, Mario; Torelli, G; Mar, E. C.
abstract

To assess the role of the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) gene in the development of Hodgkin´s lymphoma (HL), the polymorphism of this gene in EBV isolates from different geographic locations was analyzed. A 497 bp fragment spanning LMP1 gene exons 1 and 2 was amplified by polymerase chain reaction (PCR), using a primer pair bracketing a Xhol restriction site. PCR products were subjected to Xhol digestion and to DNA sequencing analysis. Twenty-five HL biopsy specimens from the United States and five HL and four non-Hodgkin´s lymphoma (NHL) biopsy specimens from Italy were examined. Eighty percent of LMP1-positive samples (12 of 15) from the United States maintained the Xhol restriction site and the remaining 20% partially lost the Xhol site. One of four EBV-positive HL and one of the three EBV-positive NHL specimens from Italy lost the restriction site. The other three EBV-positive HL DNAs were partially cut by Xhol. Direct DNA sequencing analysis revealed that those Italian samples not digested by Xhol were due to a G to C transversion at the first base of codon 18, resulting in the change of glycine to arginine. Those DNA samples partially cut by Xhol were due to a mixture of G/C at the same location. In contrast, those partially digested American HL DNAs had a mixture of GTT at the second base of codon 17. The sequence variation found in the Italian samples differs from that of Asian EBV strains, in which G to T transversion was detected at codon 17, resulting in the substitution of arginine by leucine. Among the 72% (18 of 25) EBV-positive American HL samples, 67% (12 of 18) were associated with type A virus, 17% (3 of 18) with type B, and 17% (3 of 18) with dual viral sequences. EBV DNA was detected in 80% (four of five) of Italian HL biopsy specimens, in which 50% (two of four) were associated with type A and 50% (two of four) with type B. Despite these sequence variations at the Xhol recognition site between EBV isolates of different geographic locations, no direct correlation with a specific genotype was observed. These results, to our knowledge, represent the first observation of a specific point mutation at codon 18 of LMP1 gene associated with a particular geographic location. It appears that the Xhol polymorphism may be a useful molecular marker for epidemiologic study, and the alteration in the LMP1 gene may have functional significance in the development of HL in certain geographic areas.

1995 - HUMAN HERPESVIRUS-6 - A SURVEY OF PRESENCE AND DISTRIBUTION OF GENOMIC SEQUENCES IN NORMAL BRAIN AND NEUROGLIAL TUMORS [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Maiorana, Antonino; Marasca, Roberto; R., Trovato; Fano, Rita Adriana; L., CECCHERINI NELLI; Torelli, Giuseppe
abstract

In an attempt to study the frequency and distribution of human herpesvirus-6 (HHV-6) infection both in normal and neoplastic brain tissues in vivo, polymerase chain reaction was used to look for HHV-6 genomes: 1) in samples, obtained at necropsy, from different regions of the brain of immunocompetent adult subjects and of patients who died of AIDS; 2) in the surgical biopsies of a well-characterized series of primary brain tumors of neuroglial origin. HHV-6-specific sequences were identified in six of nine brain samples from immunocompetent subjects, and in four of seven brain samples from AIDS patients. Viral sequences were identified in the specimens derived either from the grey (frontal cortex and basal ganglia) or from the periventricular white matter. HHV-6 DNA was found only in 6 of the 37 primary brain tumor biopsies examined. Th is study provides for the first time molecular evidence of a wide distribution of HHV-6 infection in the brain tissues of a high proportion of subjects, both in normal and in impaired immunity. in this large series of tumor biopsies the presence of HHV-6 genomic sequences is a rare phenomenon, arguing against a major role of this herpesvirus in the pathogenesis of primary brain tumors of neuroglial origin in immuno-competent subjects.

1995 - Hairy cell leukaemia, second cancer and occupational risk [Articolo su rivista]
Emilia, G; Luppi, Mario; Gandini, G; Bertesi, M; Torelli, G.
abstract

No abstract available

1995 - Human herpesvirus-6 (HHV-6) in blood donors [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Marasca, Roberto; Ceccherini Nelli, L; Ceccherelli, G; Torelli, G.
abstract

No abstract available

1995 - Human herpesvirus-6 genome in acute lymphoblastic leukemia: Evidence against an etiologic relationship [Articolo su rivista]
Barozzi, Patrizia; Luppi, Mario; Marasca, Roberto; Trovato, R; Ceccherininelli, L; Torelli, Giuseppe
abstract

No abstract available

1995 - Logopedic rehabilitation of laryngeal granulomas [Articolo su rivista]
Luppi, Mario
abstract

Posterior laryngeal granuloma is an infrequent pathology of multidisciplinary interest. Actually, its real prevalence is difficult to quantify because in some cases it is asymptomatic and in other instances it may either be reabsorbed or eliminated spontaneously. It is located at the vocal apophysis of the arytenoid or, less frequently, above it or on the laryngeal side of the arytenoid. The many etiologic factors (laryngeal intubation, gastro-esophageal refluxes, blunt trauma of the larynx, vocal dysfunction), sometimes concomitant and with the possible addition of enhancing circumstances (upper aerodigestive tract inflammation, naso-gastric tube, smoking and alcohol abuse), converge to a single pathogenetic mechanism: an ulceration of the mucosa and the pericondrium, sometimes complicated by an infection, which does not heal but instead produces a typical granulation tissue with capillaries oriented radially from the center of the lesion. Post intubation granulomas, extremely rare in children, are more frequent in females. It appears that there is no correlation with duration of intubation in that granulomas, can also occur after short general anesthesia. Idiopathic or contact granulomas are more frequent in the males. They are the result of vocal laryngeal hyperfunction, habitual throat clearing or cough-like throat clearing. Gastro-esophageal reflux of gastric juice, coughing or throat clearing may injure the mucosa. A blunt trauma of the larynx may cause a granuloma if the cartilage of the vocal process is exposed. Symptoms, when present, are dysphonia, tiredness during or after voicing, bolus, laryngeal unilateral pain, sensation of something in the throat which is mobile during breathing and swallowing, traces of blood in the expectoration. Therapeutic options are surgical, medical or logopedic. Surgery, although followed by frequent recurrences, is mandatory when the granuloma causes dispnea or if a pathologic essay is needed. Medical treatment aims at solving gastroesophageal reflux and/or inflammations of the district. Logopedic rehabilitation is the most successful therapy. Since January 1992 the Authors have been adopting the rehabilitation protocol planned by the French phoniatrician Brigitte Arnoux-Sindt for post-intubation granulomas, which, moreover, is utilyzed for all type of granulomas, including those arising during the early postoperative period after cordectomy. This protocol is analytically presented and discussed. In the cases of contact granulomas, and when there is concomitant vocal dysfunction, logopedic treatment is prolonged after granuloma dissapearance with some sessions aiming at restoring correct vocal behaviour. In all the ten patients rehabilitated up to now, granulomas disappeared after a mean of 16.3 sessions held twice a week. After several months of follow-up we had no recurrences. This clinical experience, while limited in number, seems to confirm the good results already reported in French Literature.

1995 - Spontaneous adrenal gland haematoma in a patient with antiphospholipid antibodies [Articolo su rivista]
Luppi, Mario; Marasca, Roberto; Torricelli, Pietro; Leonardi, G; Gavioli, Gl; Bocchi, A; Torelli, G.
abstract

No abstract available

1995 - Spontaneous loss of Ph chromosome with maintenance of clonal hemopoiesis in an untreated patient with myeloproliferative disease and a long survival. [Articolo su rivista]
Luppi, Mario; Morselli, M; Emilia, G; Temperani, P; Marasca, Roberto; Barozzi, Patrizia; Selleri, L; Torelli, G.
abstract

The unusual case of myeloproliferative disease described here is characterized by the following features: (1) a clinically completely silent course for 11 years without splenomegaly, marrow fibrosis, or cellular morphologic alterations; (2) the presence, at the onset, of a Philadelphia (Ph) chromosome without DNA breakpoints in the M-bcr region; (3) the spontaneous loss of detectable Ph-positive cells, 5 years after the first finding of leukocytosis, in the absence of any therapy; (4) the maintenance of the clonal nature of hematopoiesis, as revealed by the PGK X-linked inactivation pattern, in the absence of the Ph chromosome; and (5) a biphasic trend in the levels of leukocytes, red cells, and platelets during the years of observation.

1995 - THE USE OF CYCLOSPORINE-A IN THE TREATMENT OF B-CHRONIC LYMPHOCYTIC-LEUKEMIA - replay [Articolo su rivista]
Emilia, G; Messora, C; Bensi, L; Luppi, Mario
abstract

No abstract available

1995 - The use of cyclosporin-A in the treatment of B-chronic lymphocytic leukemia [1] [Articolo su rivista]
Luppi, Mario
abstract

1994 - Clonal nature of hypereosinophilic syndrome [Articolo su rivista]
Luppi, Mario; Marasca, Roberto; Morselli, M; Barozzi, Patrizia; Torelli, Giuseppe
abstract

No abstract available

1994 - DOUBLE P53 POINT MUTATION IN EXTRAMEDULLARY BLAST CRISIS OF CHRONIC MYELOGENOUS LEUKEMIA [Articolo su rivista]
Marasca, Roberto; Longo, G; Luppi, Mario; Barozzi, Patrizia; Torelli, Giuseppe
abstract

A patient with Philadelphia-positive chronic myelogenous leukemia (CML) evolved in extra-medullary blast crisis, was studied for the presence of alterations of the P53 tumor suppressor gene in the different stages of disease progression. No P53 gene aberrations were detected during the chronic and accelerated phases. Two identical missense point mutations, involving codons 249 and 281 and leading to the amino acid substitutions Arg-Ser and Thy-Asp, were identified in cells of an extramedullary mass and then in peripheral blood blast crisis cells. The data indicate that the medullary and extramedullary blast cells belong to the same cellular clone. They also strongly suggest that in this case, the alteration of P53 gene is strictly related to the progression of the disease, although this mechanism is certainly neither the only nor the most frequent molecular event leading to the acute phase.

1994 - Human herpesvirus 6 infection in normal human brain tissue [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Maiorana, Antonino; Marasca, Roberto; Torelli, Giuseppe
abstract

No abstract available

1994 - INTEGRATION OF HUMAN HERPESVIRUS-6 (HHV-6) GENOME IN CHROMOSOME-17 IN 2 LYMPHOMA PATIENTS [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Marasca, Roberto; Torelli, Giuseppe
abstract

No abstract available

1994 - The human A-myb protein is a strong activator of transcription [Articolo su rivista]
Golay, J; Loffarelli, L; Luppi, Mario; Castellano, M; Introna, M.
abstract

The A-myb gene belongs to the family of the c-myb proto-oncogene. We report here the cloning from a B lymphocyte cDNA library of the previously missing 3' half of the human A-myb cDNA, thus closing the previously still incomplete open reading frame. Analysis of the homologies between the different myb proteins reveals four domains of high conservation. We show, using a polyclonal rabbit antibody, that the 90 kd human A-myb protein is nuclear and that it activates transcription from the KHK-CAT reporter 6-10 times more strongly than c-myb in NIH3T3 cells. The transactivating function of A-myb depends on the presence of the myb binding site in the reporter, and on both the DNA binding and acidic domains of the A-myb protein. The bacterially expressed protein protects the myb binding sites of the reporter in footprint experiments. Binding of the A-myb protein is shown in gel retardation assays to be specific for the classical c-myb recognition sequence PyAACG/TG. In addition, like c-myb, A-myb binds more strongly to the MIM-A synthetic oligonucleotide that carries the TAACGG sequence than to the MBS-I oligonucleotide containing TAAGTG. Finally, DNA binding activity is demonstrated to require the N-terminal portion of the protein containing the three tandem repeats of amino acids conserved in all myb proteins. We have thus shown that the A-myb protein is a strong activator of transcription and that this activity depends on both the DNA-binding and acidic domains.

1993 - 3 CASES OF HUMAN HERPESVIRUS-6 LATENT INFECTION - INTEGRATION OF VIRAL GENOME IN PERIPHERAL-BLOOD MONONUCLEAR CELL-DNA [Articolo su rivista]
Luppi, Mario; Marasca, Roberto; Barozzi, Patrizia; Ferrari, Sergio; Ceccherininelli, L; Batoni, G; Merelli, E; Torelli, Giuseppe
abstract

Saliva and peripheral blood mononuclear cells from three patients, two with lymphoproliferative disorders and one suffering from multiple sclerosis, were examined for the presence of human herpesvirus-6 (HHV-6) genome by using the polymerase chain reaction and Southern blot analysis. The search for anti-HHV-6 antibodies, carried out in the sera of the same cases by an immunofluorescence assay, was negative in two cases at the lowest dilution used (1:40). These three patients had a high number of HHV-6 specific sequences in uncultured peripheral blood mononuclear cells, which are thought to be a normal site of viral latency although, in healthy individuals, the infected cells are extremely rare. In order to gain some insight into the state of the viral genome in this latent HHV-6 infection, we used pulsed field gel electrophoresis to separate HHV-6 DNA directly from HHV-6 (strain GS) infected HSB-2 cells and from the peripheral blood mononuclear cells of these three patients. Our study showed the presence of intact viral genome, of the expected length of 170 kb, persisting as free extrachromosomal element in the HSB-2 cells but not in patients' peripheral blood mononuclear cells. On the other hand, in strong contrast with the results obtained in infected HSB-2 DNA, the restriction analysis of the three patients' peripheral blood mononuclear cell DNA showed fragments of molecular weight constantly higher than the 170 kb segment, indicating that the viral sequences are linked to high molecular weight cellular DNA. Our findings are consistent only with a latent infection in which HHV-6 is integrated in vivo and suggest that pulsed field gel electrophoresis analysis is well worth using to evaluate the presence of integrated, intact, or fragmented viral genomes in HHV-6 associated lymphoproliferative diseases and immune disorders.

1993 - CHARACTERIZATION OF HUMAN HERPESVIRUS-6 GENOMES FROM CASES OF LATENT INFECTION IN HUMAN LYMPHOMAS AND IMMUNE DISORDERS [Articolo su rivista]
Luppi, Mario; Barozzi, Patrizia; Marasca, Roberto; Ceccherininelli, L; Torelli, Giuseppe
abstract

No abstract available

1993 - Comparative study of famotidine vs ranitidine in critically ill patients in mechanical ventilation [Articolo su rivista]
Luppi, Mario
abstract

Stress ulcers are frequent in critically ill patients. Gastric mucosa protection with antacids, H2 blockers or sucralphate decrease the incidence of stress ulcers and upper gastrointestinal bleeding but may have adverse effects. The present study compares the efficacy of continuous infusions of famotidine on gastric pH control and bleeding prevention in critically ill patients receiving mechanical ventilation. Fifty six patients (aged 55 +/- 22 years, 26 male) were randomly assigned to receive famotidine 40 mg/day (n = 27) or ranitidine 150 mg/day (n = 29) during 5 days. Gastric pH was measured every 6 hours and when it was below 5, aluminum hydroxide (30 ml every 2 hours) was administered to obtain values over 5. Patients receiving famotidine had higher mean gastric pH (6.3 +/- 0.2 and 93% of measurements over 5 vs 5.8 +/- 0.6 and 83% of measurements over 5 (p &lt; 0.05), and required lower amounts of aluminum hydroxide (1.880 vs 2.770 ml). No patient had evidence of gastrointestinal bleeding and one had a mild psychomotor agitation. No other adverse reactions were observed. It is concluded that famotidine was more effective than ranitidine on gastric pH control in mechanically ventilated critically ill patients.

1993 - Detection of high levels of human herpes virus-6 DNA in a lymphoma of a patient with Sjögren's syndrome. [Articolo su rivista]
Fox, Ri; Luppi, Mario; Kang, Hi; Ablshi, D; Josephs, S.
abstract

No abstract available.

1993 - FREQUENT DETECTION OF HUMAN HERPESVIRUS-6 SEQUENCES BY POLYMERASE CHAIN-REACTION IN PARAFFIN-EMBEDDED LYMPH-NODES FROM PATIENTS WITH ANGIOIMMUNOBLASTIC LYMPHADENOPATHY AND ANGIOIMMUNOBLASTIC LYMPHADENOPATHY-LIKE LYMPHOMA [Articolo su rivista]
Luppi, Mario; Marasca, Roberto; Barozzi, Patrizia; Artusi, T; Torelli, Giuseppe
abstract

In search of a possible involvement of viral agents in angioimmunoblastic lymphadenopathy with dysproteinaemia (AILD) and AILD-like lymphoma (AILD-L), we studied the presence of human herpesvirus-6 (HHV-6) in the lymph node biopsies of 12 cases by polymerase chain reaction (PCR). Given the rarity of this lymphoproliferative disorder, we investigated archival specimens, consisting of formalin-fixed and paraffin-embedded tissues, obtained from patients with a clinical and histologic diagnosis of AILD and AILD-L. HHV-6 sequences were detected in the lymph node biopsies of 7 out of the 12 AILD and AILD-L cases examined. HHV-6 sequences were identified also in the involved liver and spleen tissues of one patient and in the PBMCs of two patients, all carrying viral sequences in the affected lymph nodes. We also used PCR to characterize the HHV-6 genomes, showing that two different viral strains are represented in the pathologic tissues. This study provides evidence of the presence of HHV-6 specific sequences in an unexpectedly high proportion of our series of AILD and AILD-L cases, suggesting a possible involvement of this lymphotropic virus in the pathogenesis of at least some cases of the disease.

1993 - HUMAN HERPESVIRUS-6 AND MULTIPLE-SCLEROSIS - SURVEY OF ANTI-HHV-6 ANTIBODIES BY IMMUNOFLUORESCENCE ANALYSIS AND OF VIRAL SEQUENCES BY POLYMERASE CHAIN-REACTION [Articolo su rivista]
Sola, P; Merelli, E; Marasca, Roberto; Poggi, M; Luppi, Mario; Montorsi, M; Torelli, Giuseppe
abstract

A possible involvement of human herpesvirus 6 (HHV-6) infection in the pathogenesis of multiple sclerosis (MS) was investigated. The immunofluorescence analysis of sera from 126 MS patients showed significantly higher anti-HHV-6 antibody titres in MS sera than in 500 normal controls. A polymerase chain reaction (PCR) assay of the peripheral blood mononuclear cell (PBMC) DNAs of 31 MS patients and 24 normal subjects was positive in one normal control and in one MS patient. The Southern blot analysis indicated an unexpectedly high level of viral sequences in the MS patient, but not in the control. Since viral sequences are rarely present in MS subjects, the high anti-HHV-6 antibody titres found in MS are likely to be related to immune impairment rather than reactivation of a latent infection.

1992 - HUMAN HERPESVIRUS-6 IN NON-AIDS RELATED HODGKINS AND NON-HODGKINS-LYMPHOMAS [Articolo su rivista]
Torelli, Giuseppe; Marasca, Roberto; Montorsi, M; Luppi, Mario; Barozzi, Patrizia; Ceccherini, L; Batoni, G; Bendinelli, M; Muyombano, A.
abstract

Human herpesvirus 6 in non-AIDS related Hodgkin's and non-Hodgkin's lymphomas.

1992 - In vitro transformation by HHV-6 (Human Herpes-virus 6) [Articolo su rivista]
G., Batoni; R., Trovato; F., Lojacono; L. A., Barsanti; M., Morelli; P., Barozzi; Luppi, Mario; Maiorana, Antonino; Torelli, Giuseppe; L., Ceccherini Nelli
abstract

The in-vitro transformation by human herpesvirus 6 is described

1992 - Potential role of Epstein-Barr virus in Sjögren's syndrome and rheumatoid arthritis. [Articolo su rivista]
Fox, Ri; Luppi, Mario; Pisa, P; Kang, H. I.
abstract

In the pathogenesis of Sjögren's syndrome (SS), a role for Epstein-Barr virus (EBV) has been suggested because; (a) EBV is present in salivary gland epithelial cells of healthy individuals, and exaggerated immune responses against EBV could play a role in the destruction of salivary glands in SS; (b) SS salivary gland biopsies contain increased levels of EBV DNA compared to normal salivary glands, indicating viral reactivation and inability of lymphoid infiltrates to control EBV replication in patients with SS; and (c) salivary gland epithelial cells in patients with SS express high levels of HLA-DR antigens and may present EBV associated antigens to immune T cells in patients with SS. Therefore, SS may represent a situation where genetically predisposed individuals (i.e., HLA-DR3-DQA4-DQB2) have a persistent but ineffectual T cell immune response against EBV at its site of latency. In the case of rheumatoid arthritis (RA), evidence for a potential role of EBV includes the following: (a) EBV encoded proteins share antigenic and sequence similarity to proteins found in the synovial tissues. These crossreactive proteins include EBV protein gp110 (BALF-4) and the beta chain of HLA-DR4. Also, the human and mycobacterial 65 kDa heat shock proteins have a sequence similar to that of EBV encoded proteins; (b) patients with RA have increased frequency and levels of antibodies against specific epitopes on EBV encoded EBNA-1 (BKRF-1) and EBNA-3 (BERF-1) antigens; and (c) lymphocytes from patients with RA have decreased ability to limit outgrowth of autologous EBV infected lymphocytes, probably due to defects in release of interferon gamma.

1991 - HHV-6 INFECTION AND HUMAN HODGKIN AND NON-HODGKIN LYMPHOMAS [Articolo su rivista]
Torelli, Giuseppe; Marasca, Roberto; Selleri, L; Luppi, Mario; Montorsi, M; Federico, Massimo; Narni, Franco; Ceccherininelli, L; Ferrari, Sergio
abstract

1991 - Human Herpes virus-6 in human lymphomas: identification of specific sequences in Hodgkin's lymphomas by polymerase chain reaction [Articolo su rivista]
Torelli, Giuseppe; Marasca, Roberto; Luppi, Mario; L., Selleri; Ferrari, Sergio; Narni, Franco; Mt, Mariano; Federico, Massimo; L., CECCHERINI NELLI; M., Bendinelli; G., Montagnani; M., Montorsi; Artusi, Tullio
abstract

In search of a possible involvement of the human herpesvirus type 6 (HHV-6) in human Hodgkin's and non-Hodgkin's lymphomas, we studied the levels of anti-HHV-6 antibodies in the sera of 94 cases by an immunofluorescence assay, as well as the presence of HHV-6 sequences in the affected tissues of 66 cases by polymerase chain reaction, using one set of primer oligonucleotides. Our results showed higher anti-HHV-6 antibody titers in human lymphomas than in normal blood donors, but the difference is statistically significant only when normal donors are compared with Hodgkin's lymphoma cases. HHV-6 sequences were detected in 3 of 25 Hodgkin's lymphomas and 0 of the 41 cases of non-Hodgkin's lymphomas studied. The three cases positive for HHV-6 sequences belong to the nodular sclerosis-lymphocyte depletion histologic subtype and share remarkable similarities in their clinical features. Furthermore, Southern blot analysis of total genomic DNA obtained from the neoplastic tissues of two of the three patients showed the same restriction fragment length polymorphism. Our results suggest that: (1) the high level of anti-HHV-6 antibodies in Hodgkin's disease is due to an activation of the immune system not related to the presence of HHV-6 sequences in affected lymph nodes; (2) the presence of HHV-6 sequences in human lymphoid tissues is not a frequent event, rather it is in fact a very rare event in non-Hodgkin's lymphomas, while in Hodgkin's cases it is more frequent than previously reported on the basis of Southern blot analysis; and (3) the presence of HHV-6 sequences in Hodgkin's lymphomas may have a relation with the clinical presentation of the disease.

1991 - Reactivation of Epstein-Barr virus in Sjögren's syndrome. [Articolo su rivista]
Fox, Ri; Luppi, Mario; Kang, Hi; Pisa, P.
abstract

Sjögren's syndrome (SS) is a chronic autoimmune disease characterized by severe dryness of the eyes and mouth, resulting from lymphocytic infiltration of the lacrimal and salivary glands. SS may exist as a primary condition (primary SS, 1.SS) or as a secondary condition (2.SS) in association with rheumatoid arthritis, systemic lupus erythematosus, or progressive systemic sclerosis. In some 1.SS patients, there may be involvement of the extraglandular organs, including skin, kidney, liver, lung and nervous system. Furthermore, these patients may develop a lymphoproliferative syndrome that includes lymphadenopathy and increased risk of lymphoma. In the pathogenesis of SS, a role for Epstein-Barr virus (EBV) has been suggested because: (a) EBV is present in salivary gland epithelial cells of normal individuals and exaggerated immune responses against EBV could play a role in the destruction of salivary glands in SS; (b) SS salivary gland biopsies contain increased levels of EBV DNA in comparison to normal salivary glands, indicating viral reactivation and inability of lymphoid infiltrates to control EBV replication in SS patients; and (c) salivary gland epithelial cells in SS patients express high levels of HLA-DR antigens and may present EBV-associated antigens to immune T cells in SS patients. Therefore, SS may represent a situation in which genetically predisposed individuals (i.e., HLA-DR3-DQA4-DQB2) have a persistent but ineffectual T cell immune response against EBV at its site of latency. Among 14 non-Hodgkin's lymphomas that developed in SS patients, EBV DNA was detected in increased amounts in the tumor tissue of one patient. Characterization of this tumor DNA revealed: (a) polyclonal immunoglobulin gene rearrangements; (b) EBV DNA with an unusual restriction fragment length polymorphism pattern involving the Bam M fragment; and (c) EBV terminal repeat sequences suggestive of viral replication, similar to those reported in EBV lymphomas occurring in other immunocompromised individuals. Early recognition of this clinical problem may allow beneficial use of antiviral agents.

1990 - Chronic myelogenous leukemia with typical clinical and morphological features can be Philadelphia chromosome negative and "bcr negative". [Articolo su rivista]
Selleri, L; Emilia, Giovanni; Luppi, Mario; Temperani, Paola; Zucchini, Patrizia; Tagliafico, Enrico; Artusi, Tullio; Sarti, M; Donelli, A; Castoldi, Gl
abstract

The Philadelphia (Ph1) chromosome is found in the majority of patients affected by chronic myelogenous leukemia (CML), being considered the hallmark of the disease, but around 5-8% of patients diagnosed as CML lack the Ph1 chromosome-negative (Ph-) CML has been discussed extensively in the literature because of its heterogeneity. However, it is now accepted that some of the Ph1-CML patients have a disease indistinguishable from Ph1-positive (Ph+) CML. It was investigated whether Ph- CML with clinical and morphological features indicating true CML would always have bcr rearrangements, as the relocation of c-abl from 9q34 into the breakpoint cluster region on 22q11 is considered a crucial event in the pathogenesis of CML. From molecular studies, it seemed that Ph- CML with features of true CML always have the bcr rearrangement, while Ph- patients, lacking such rearrangement, have atypical forms of CML. Here we describe 8 Ph- CML and myeloproliferative syndrome (MPS) patients of whom 6 were by all respects true CML cases. Nevertheless, bcr rearrangement and expression of the classic bcr/abl chimeric mRNA was found in only 1 of the 6 patients. More advanced molecular techniques will be needed to understand which molecular mechanisms underlie Ph-, bcr- CML, resulting in phenotypes sometimes indistinguishable from Ph+, bcr+ CML.

1990 - IDENTIFICATION OF SEQUENCES OF HUMAN HERPES VIRUS-6 (HHV-6) IN A CASE OF HODGKINS-DISEASE BY POLYMERASE CHAIN-REACTION [Articolo su rivista]
Marasca, Roberto; Luppi, Mario; Montorsi, M; Fancinelli, M; Sabbatini, R; Mariano, Mt; Selleri, L.
abstract

In the last decades new approaches to the diagnosis and treatment of Hodgkin's Disease (HD) have contributed to improved rates of survival and cure but the pathogenesis of the disease still remains unknown. Data have been collected suggesting a relationship between viral infections and HD. HD patients with evidence of Epstein-Barr virus genomes in their affected tissues have been recently reported. Human Herpes Virus six (HHV-6) is a newly isolated virus, derived from patients with lymphoproliferative disorders. In order to investigate the possible role of this virus in the pathogenesis of HD we looked for a specific segment of HHV-6 genome by means of the polymerase chain reaction (PCR) in tissue samples obtained from peripheral blood and lymphnodes in HD patients: one clearly positive case has been identified. This result is the first indication of HHV-6 sequences associated with a case of HD and raises the possibility that this virus might be involved in the pathogenesis of this lymphoproliferative disorder. The relationship between HHV-6 and HD therefore warrants further investigation.

1990 - MITOCHONDRIAL-DNA DELETION IN A PATIENT WITH PROGRESSIVE OPHTHALMOPLEGIA [Articolo su rivista]
Luppi, Mario; Marasca, Roberto; Sola, P; Corradi, M; Fancinelli, M; Montorsi, M; Manfredini, Rossella; Selleri, L.
abstract

Mitochondria are unique among intracellular organelles because they contain their own DNA, which can be transcribed and translated to form proteins. Mitochondrial diseases include myopathies and multisystem disorders. The case of a patient showing bilateral ophthalmoplegia with proximal limb weakness, severe dysphagia and short stature, without family history, is described. The analysis of mitochondrial DNA of the patient muscle revealed a deleted form accounting for 65% of the total mitochondrial DNA. The Southern Blot Analysis of mtDNA allows a rather precise localization of deletions giving new insights in the pathogenesis of mitochondrial myopathies and representing a new precious diagnostic tool in these diseases.

1990 - SURVEY OF THE PRESENCE OF ANTI-HHV6 ANTIBODIES AND HHV6 SEQUENCES IN NON-HODGKIN LYMPHOMA CASES [Articolo su rivista]
Selleri, L; Marasca, Roberto; Luppi, Mario; Montorsi, M; Ceccherininelli, L; Bendinelli, M; Torelli, Giuseppe
abstract

1989 - Polymerase chain reaction for the diagnostic identification of HIVinfection [Articolo su rivista]
Selleri, L; Grassilli, E; Tagliafico, Enrico; Corradi, M; Luppi, Mario; Ceccherelli, G; Borghi, V.
abstract

The Acquired Immune deficiency Syndrome (AIDS), caused by the Human Immunodeficiency Virus (HIV) is now a worldwide social problem. Routine diagnostic procedures to identify infected individuals are based on the presence of antibodies against viral epitopes in the serum. There is nevertheless impelling need to detect directly the virus in people infected by HIV, independently of a serological response. In this study we describe the procedure which allows amplification of a specific segment of the HIV genome, through the Polymerase Chain Reaction (PCR), in infected individuals. This new approach represents a precious tool towards the diagnosis of HIV infection, it can be easily and quickly carried out on a large scale and will be capable of identifying HIV infected subjects before the development of antibodies.