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STEFANIA BENATTI

COLLABORATORE COORDINATO CONTINUATIVO
Dipartimento di Scienze Mediche e Chirurgiche Materno-Infantili e dell'Adulto

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Pubblicazioni

2022 - Multicentre match-paired analysis of advanced biliary cancer long-term survivors: The BILONG study [Articolo su rivista]
Caputo, F.; Gelsomino, F.; Spallanzani, A.; Pettorelli, E.; Benatti, S.; Ghidini, M.; Grizzi, G.; Ratti, M.; Merz, V.; Messina, C.; Tonelli, R.; Luppi, G.; Melisi, D.; Dominici, M.; Salati, M.
abstract

Background: Advanced biliary cancers (ABC) are aggressive malignancies with a median overall survival (mOS) <12 months when treated with first-line chemotherapy. Nevertheless, a subset of patients experiencing longer survival has been described in the updated analysis of ABC-02 trial. We aimed to provide a real-world description of ABC long-term survivors (LS), identifying which factors impact on survival. Methods: Patients diagnosed with ABC at three Institutions between 2002 and 2019, and who survived ≥18 months, were retrospectively identified. We compared them with a control cohort (C) with a mOS <18 months, matched on age, gender, ECOG PS, disease status, primary tumor site, prior surgery, and treatment modality. Their clinical features, treatments, and outcome were analyzed. Results: A total of 78 patients was included, 39 in each group. Both LS and C cohorts had superimposable baseline characteristics, without significant differences. mOS was 29 (95%CI 24.6–33.5) and 9 months (95%CI 6.6–12.9) in the two groups, respectively. After performing a logistic regression analysis, three factors were significantly associated with long-term outcome: low neutrophil-to-lymphocyte ratio (NLR < 3) (Odds Ratio [OR] 0.38), achievement of objective response to treatment (OR 0.16), and the number of lines received (OR 0.29). Conclusions: We described a considerable subset of ABC experiencing long-term survival with conventional chemotherapy in a real-world scenario. Beyond clinical factors, we identified low NLR as a prognostic determinant that may allow for a more accurate selection of long survivors. While waiting for a deeper molecular characterization of this subgroup, we propose NLR as a stratification factor for daily practice and clinical trials.

2022 - Statins increase pathological response in locally advanced rectal cancer treated with chemoradiation: a multicenter experience. [Articolo su rivista]
Caputo, F; Santini, C; Casadei-Gardini, A; Cerma, K; Bardasi, C; Garajovà, I; Lattanzi, E; Passardi, A; Rapposelli, Ig; Spallanzani, A; Salati, M; Bonetti, Lr; Gelmini, R; Meduri, B; Piccoli, M; Pecchi, A; Benatti, S; Piacentini, F; Dominici, M; Luppi, G; Gelsomino, F.
abstract

Aims: To investigate the influence of various concomitant medications on outcomes in patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiation. Materials & methods: The authors retrospectively identified 246 patients from 2003 to 2018, collecting demographic and clinicopathological data of interest. Odds ratio (OR) was used to assess the association between concomitant drugs and outcomes. Results: The authors found an association between statins and a Dworak regression grade of 3-4 (OR = 8.78; p = 0.01). Furthermore, statins were significantly associated with more frequent chemoradiation-related toxicity (OR = 2.39; p = 0.0098) and chemotherapy dose reduction or discontinuation (OR = 2.26; p = 0.03). Conclusion: Despite higher frequency of radiotherapy and chemotherapy interruption or dose reduction, the concomitant use of statins during neoadjuvant chemoradiation proved to be associated with better tumor regression.

2021 - The Prognostic Role of Early Skeletal Muscle Mass Depletion in Multimodality Management of Patients with Advanced Gastric Cancer Treated with First Line Chemotherapy: A Pilot Experience from Modena Cancer Center [Articolo su rivista]
Rimini, M; Pecchi, A; Prampolini, F; Bussei, C; Salati, M; Forni, D; Martelli, F; Valoriani, F; Canino, F; Bocconi, A; Gelsomino, F; Reverberi, L; Benatti, S; Piacentini, F; Menozzi, R; Dominici, M; Luppi, G; Spallanzani, A
abstract

Background: Few data about the link between nutritional status and survival are available in the metastatic gastric cancer (GC) setting. The aim of this work was to evaluate the prognostic role of tissue modifications during treatment and the benefit of a scheduled nutritional assessment in this setting. Methods: Clinical and laboratory variables of 40 metastatic GC patients treated at Modena Cancer Center were retrieved: 20 received a nutritional assessment on the oncology’s discretion, the other 20 received a scheduled nutritional assessment at baseline and every 2–4 weeks. Anthropometric parameters were calculated on Computed Tomography (CT) images at the baseline and after 3 months of chemotherapy. Results: A correlation between baseline Eastern Cooperative Oncology Group Performance Status (ECOG PS), Lymphocyte to Monocyte Ratio (LMR), C-reactive protein (PCR), Prognostic Nutritional Index (PNI) and Overall survival (OS) was highlighted. Among the anthropometric parameters, early skeletal muscle mass depletion (ESMMD) >10% in the first months of treatment significantly impacted on mOS (p = 0.0023). A link between ESMMD and baseline LDH > 460 U/L, baseline CRP > 2.2 mg/dL and weight decrease during treatment emerged. Patients evaluated with a nutritional scheduled support experienced a mean gain in subcutaneous and visceral fat of 11.4% and 10.21%, respectively. Conclusion: We confirm the prognostic impact of ESMMD > 10% during chemotherapy in metastatic GC. The prognostic role of a scheduled nutritional assessment deserves further confirmation in large prospective trials.

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 - 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.

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

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 - 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 - 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.

2017 - A prokineticin-like protein responds to immune challenges in the gastropod pest Pomacea canaliculata [Articolo su rivista]
Accorsi, Alice; Benatti, Stefania; Ross, Eric; Nasi, Milena; Malagoli, Davide
abstract

The golden apple snail Pomacea canaliculata is an invasive pest originating from South America. It has already been found in Asia, the southern United States and more recently in the EU. Aiming to target the immune system of the snail as a way to control its spreading, we have developed organ-specific transcriptomes and looked for molecules controlling replication and differentiation of snail hemocytes. The prokineticin domain-containing protein Astakine 1 is the only cytokine known thus far capable of regulating invertebrate hematopoiesis, and we analyzed the transcriptomes looking for molecules containing a prokineticin domain. We have identified a prokineticin-like protein (PlP), that we called Pc-plp and we analyzed by real-time PCR (qPCR) its expression. In control snails, highest levels of Pc-plp were detected in the digestive gland, the ampulla (i.e., a hemocyte reservoir) and the pericardial fluid (i.e., the hematopoietic district). We tested Pc-plp expression after triggering hematopoiesis via multiple hemolymph withdrawals, or during bacterial challenge through LPS injection. In both cases a reduction of Pc-plp mRNA was observed. The multiple hemolymph withdrawals caused a significant decrease of Pc-plp mRNA in pericardial fluid and circulating hemocytes, while the LPS injection promoted the Pc-plp mRNA drop in anterior kidney, mantle and gills, organs that may act as immune barrier in molluscs. Our data indicate an important role for prokineticin domain-containing proteins as immunomodulators also in gastropods and their dynamic expression may serve as a biosensor to gauge the effectiveness of immunological interventions aimed at curtailing the spreading of the gastropod pest P. canaliculata.

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 - 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.

2016 - Changes in the NMR Metabolic Profile of Live Human Neuron-Like SH-SY5Y Cells Exposed to Interferon-α2 [Articolo su rivista]
Valeria, Righi; Schenetti, Luisa; Mucci, Adele; Benatti, Stefania; Tascedda, Fabio; Brunello, Nicoletta; Carmine, Pariante M; Alboni, Silvia
abstract

Interferon (IFN)-α2 is an extensively therapeutically used pro-inflammatory cytokine. Though its efficacy in controlling viral replication and tumor cells proliferation, administration of IFN-α2 is often associated with the development of central side effects. Magnetic resonance spectroscopy studies have demonstrated that IFN-α2 administration affects brain metabolism, however the exact nature of this effect is not completely known. We hypothesized that IFN-α2 can affect metabolic activity of human neuron-like SH-SY5Y cells which possess many characteristics of neurons and represent one of the most used models for studying mechanisms involved in neurotoxicity or neuroprotection. To test our hypothesis we have characterized the metabolic signature of live SH-SY5Y, and their conditioned media, after 24 and 72 h of exposure to vehicle or IFN-α2 (100 ng/ml) by using High Resolution-Magic Angle Spinning (HR-MAS) Nuclear Magnetic Resonance (NMR) spectroscopy. Our results revealed that 1) the use of HR-MAS NMR is ideally suitable for the characterization of the metabolic profile of live cells and their conditioned media without extraction procedures; and 2) a 72 h exposure to IFN-α2 increases the level of metabolites involved in maintaining energetic (including creatine and lactate) and osmotic (such as myo-inositol, scyllo-inositol, taurine and glycerophosphorylcholine) balances in neuron-like cells and of metabolic waste products (namely lactate, ethanol and acetate), glycine and glutamine in their growth media. These results may contribute to gain more knowledge about the IFN-α2 induced effect on the brain and support the interpretation of magnetic resonance spectroscopy studies performed in humans.

2016 - Effects of LPS injection on Pc-astakine expression in the gastropod Pomacea canaliculata [Abstract in Rivista]
Benatti, Stefania; Accorsi, Alice; Nasi, Milena; Ottaviani, Enzo; Malagoli, Davide Malagoli
abstract

Astakine-1 is a prokineticin-containing factor and the first hematopoietic cytokine described in invertebrates. Astakine-1 was firstly retrieved in the freshwater crayfish Pacifastacus leniusculus, and recent experiments have demonstrated the presence of astakine-like molecules also in insects and molluscs, including the freshwater snail, Pomacea canaliculata. In control conditions Pcastakine is expressed in several organs, especially in the ampulla (reservoir of hemocytes and potential district of hemocyte maturation) and in the pericardial fluid (i.e. the hematopoietic tissue). By mean of qPCR experiments, we have analyzed the effects of the injection of 50 µg LPS on the expression of the gene Pc-astakine. Our observations indicate that 24 h after the injection, the major modification of the Pc-astakine expression was evident in the anterior kidney, a potential hemocyte reservoir, in which the expression of the gene decreased to almost undetectable level. In the pericardial fluid, ampulla and circulating hemocytes, the expression of Pc-astakine dropped to less than 50 % with respect to the sham-injected control snails. The drop in the amount of mRNA detected by qPCR could reflect an increased rate of translation and consequent degradation of the available mRNA, rather than a decrease of the transcription rate. Similarly, in the bivalve Crassostrea gigas, it has been suggested that accumulated Cg-astakine transcripts are largely translated under some environment stress, including immune stimuli. On the whole, our results indicate that the expression of Pc-astakine and the translation rate of its mRNA may be influenced by immune stimuli, and support the hypothesis that PcAstakine may be involved in Pomacea hematopoiesis and/or may have immune-related functions, as well.

2016 - Erratum to: Changes in the NMR Metabolic Profile of Live Human Neuron-Like SH-SY5Y Cells Exposed to Interferon-α2 (J Neuroimmune Pharmacol, 2015, DOI 10.1007/s11481-015-9641-x) [Altro]
Righi, Valeria; Schenetti, Luisa; Mucci, Adele; Benatti, Stefania; Tascedda, Fabio; Brunello, Nicoletta; Pariante, Carmine M.; Alboni, Silvia
abstract

Erratum

2016 - Structure and distribution of Astakine in the organs of the freshwater snail Pomacea canaliculata. [Abstract in Rivista]
Accorsi, A.; Benatti, Stefania; Gotting, K.; Ross, E.; M. a. l. a. g. o. l. i. D., Sánchez A. l. v. a. r. a. d. o. A.
abstract

The freshwater snail Pomacea canaliculata is an emerging pest in EU, and its immune system is a potential target for developing strategies of pest control. Circulating hemocytes represent the cellular component of the P. canaliculata immune system. P. canaliculata hemocytes originate in the pericardial fluid, and are maintained in the ampulla, which may act as a hemocyte reservoir. Astakine-1 is a hematopoietic cytokine first described in the crayfish Pacifastacus leniusculus, and recently described also in the insect Lygus lineolaris and in 51 the bivalve mollusc Crassostrea gigas. Bioinformatic analyses of a comprehensive P. canaliculata transcriptome demonstrated the presence of an Astakine 1-like molecule (Pc-Astakine) also in this organism. Pc-Astakine is 121 aa and contains a domain characterized by 8 cysteins with a conserved distribution pattern homologous to the vertebrate domain prokineticin. Further bioinformatic predictions suggest that the structure of Pc-Astakine may be similar to that one of P. leniusculus Astakine-1. The distribution of Pc-astakine gene expression was evaluated by qPCR. We have observed that all organs analyzed express Pcastakine at detectable levels. However, high expression levels were detected in the ampulla, pericardial fluid and circulating hemocytes. The data suggest that Pc-Astakine may have a wide range of functions, including the regulation of hematopoiesis and the modulation of inflammatory responses, as previously reported for the human Prokineticin-1.

2014 - Circulating mitochondrial DNA increases with age and is a familiar trait: implications for "inflamm-aging" [Articolo su rivista]
Pinti, Marcello; Cevenini, E; Nasi, Milena; De Biasi, Sara; Salvioli, S; Monti, Daniela; Benatti, Stefania; Gibellini, Lara; Cotichini, R; Stazi, Ma; Trenti, T; Franceschi, C; Cossarizza, Andrea
abstract

Mitochondrial components, including mitochondrial DNA (mtDNA), when released extracellularly, can act as "damage-associated molecular pattern" (DAMP) agents and cause inflammation. As many elderly people are characterized by a low-grade, chronic inflammatory status defined "inflamm-aging", we evaluated if circulating mtDNA can contribute to this phenomenon. Eight hundred and thirty-one Caucasian subjects were enrolled in the study, including 429 siblings aged 90-104 years (90+ siblings). MtDNA plasma levels increased gradually after the fifth decade of life. In 90+ subjects, mtDNA values of two members of the same sibling relationship were directly correlated, suggesting a role for familiar/genetic background in controlling the levels of circulating mtDNA. The subjects with the highest mtDNA plasma levels had the highest amounts of TNF-??, IL-6, RANTES and IL-1ra; the subjects with the lowest mtDNA levels had the lowest levels of the same cytokines. In vitro stimulation of monocytes with mtDNA concentrations similar to the highest levels observed in vivo resulted in an increased production of TNF-??, suggesting that mtDNA can modulate the production of proinflammatory cytokines. Our findings therefore show that circulating mtDNA increases with age, and can significantly contribute to the maintenance of the low grade, chronic inflammation observed in elderly people This article is protected by copyright. All rights reserved.

2014 - Successful treatment of HIV-1 infection increases the expression of a novel, short transcript for IL-18 receptor α chain [Articolo su rivista]
Nasi, Milena; Alboni, Silvia; Pinti, Marcello; Tascedda, Fabio; Benatti, Cristina; Benatti, Stefania; Gibellini, Lara; DE BIASI, Sara; Borghi, Vanni; Brunello, Nicoletta; Mussini, Cristina; Cossarizza, Andrea
abstract

The importance of interleukin (IL)-18 in mediating immune activation during HIV infection has recently emerged. IL-18 activity is regulated by its receptor (IL-18R), formed by an α and a β chain, the IL-18-binding protein, and the newly identified shorter isoforms of both IL-18R chains. We evaluated gene expression of the IL-18/IL-18R system in peripheral blood mononuclear cells from HIV+ patients. Compared with healthy donors, IL-18 expression decreased in patients with primary infection. The IL-18Rα short transcript expression was strongly upregulated by successful highly active antiretroviral therapy. HIV progression and its treatment can influence the expression of different components of the complex IL-18/IL-18R system.

2013 - Interferon alpha exposure increases the expression of the enzymes belonging to the kynurenine pathway in an in vitro model of human neurons: SH-SY5Y cells [Abstract in Rivista]
Alboni, Silvia; Benatti, Cristina; Claudia, Montanari; Benatti, Stefania; Tascedda, Fabio; Cannazza, Giuseppe; Pariante Carmine, M; Brunello, Nicoletta
abstract

The past two decades have witnessed a burgeoning area of pre-clinical and clinical research linking psychiatric illnesses – particularly major depression (MD) – to activation of the inflammatory immune system. One of the stronger evidence supporting a causal role for inflammation in leading MD comes from reports indicating that depressive symptoms frequently develop in patients undergoing immunotherapy with cytokines, such as interferon (IFN)-α, for the treatment of malignancies or chronic viral infection. Although INF-alpha- induced effects on the brain made of IFN-α a model to study the influence of pro-inflammatory cytokines in the CNS and behavior the molecular mechanisms underlying these effects are far from being fully understood. It has been proposed that IFN-α may contribute to the etiology of MD by inducing indolamine 2,3-dioxygenase (IDO) expression and thus unbalancing in the tryptophan/kynurenine metabolism toward the production of neurotoxic metabolites and\or reducing serotonin (5-HT) availability. IDO catalyzes the initial rate-limiting step in tryptophan degradation along the kynurenine pathway (KP). Kynurenine, the initial product of tryptophan degradation, is further catalysed into neurotoxic end-products through steps catalyzed by kynurenine 3-monooxygenase (KMO) and kynureninase (Kynu). However, Kynurenine can also be catabolised by kynurenine aminotransferase (KAT), into kynurenic acid, a potentially neuroptotective agent. A role for a disturbance in the equilibrium between neurotoxic/ neuropoptective end KP endproducts producing an alteration in the neuroprotective–neurodegenerative balance in the brain of patients with MD, has been proposed in the neurodegeneration hypothesis of depression. Given that we previously demonstrated that IFN-α induces toxic effects in an in vitro model of human neurons (human SH-SY5Y neuroblastoma cells) we were aim to investigate the effects of IFN-α on KP in these cells. Our studies show that IFN-α exposure increased the expression of all the kynurenergic enzymes investigated (IDO, KMO, Kynu and KAT). More particularly strongly induced the expression of IDO mRNA (more than 900 –fold) in SH-SY5Y cells. Similar effects on kynurenergic enzyme expression were also observed when SH-SY5Y cells where differentiated with all-trans retinoic acid (in presence of neurotrophic support and in serum deprived conditions). We also demonstrated that INF-α decreased 5-HT levels whereas increased the kynurenine levels in the medium of both differentiated as well not differentiated SH-SY5Y cells.

2013 - N-acetyl-cysteine prevents toxic oxidative effects induced by IFN-α in human neurons. [Articolo su rivista]
Alboni, Silvia; Gibellini, Lara; Montanari, Claudia; Benatti, Cristina; Benatti, Stefania; Tascedda, Fabio; Brunello, Nicoletta; Cossarizza, Andrea; Pariante, Carmine M.
abstract

Currently IFN-α is widely used for effective treatment of viral infections and several malignancies. However, IFN-α can cause neuropsychiatric disturbances and mental impairments, including fatigue, insomnia, depression, irritability and cognitive deficits. Molecular and cellular mechanisms leading to such side-effects are still poorly understood. Neurons seem to be an important target in mediating cellular effects induced by exposure to this cytokine, but so far little is known about IFN-α-induced effects on these cells. We have investigated the ability of IFN-α (2-100 ng/ml) to induce damage and toxicity to the human neuroblastoma SH-SY5Y cell line, commonly used for studying such phenomena, and the mechanisms underlying these effects. After 24 h treatment, IFN-α increased mitochondrial activity, whereas cell density was reduced in a dose- and time-dependent manner. This effect did not depend on reduced cell proliferation, but rather the activation of apoptosis, as revealed by an increased Bax:Bcl-2 mRNA ratio after 72-h IFN-α exposure. At this time-point, IFN-α also reduced the expression of the brain-derived neurotrophic factor gene, and induced an increase in reactive oxygen species (ROS). A co-treatment with N-acetyl-cysteine (NAC; 5 mm), a potent antioxidant and mitochondrial modulator, was able to counteract all of these IFN-α-induced effects. These findings demonstrated that IFN-α induces neurotoxicity and apoptosis that is, in part, very likely due to mitochondrial damages and production of ROS. We suggest that NAC, already tested for the treatment of psychiatric disorders, may be useful to prevent IFN-α-induced central side-effects in a safe and effective way.

2011 - EXTRACELLULAR MITOCHONDRIAL DNA PLASMA LEVELS INCREASE DURING AGING AND LONGEVITY: IMPLICATIONS FOR "INFLAMMAGING" [Abstract in Rivista]
Cossarizza, Andrea; Cevenini, E.; Stazi, M. A.; Cotichini, R.; Monti, Daniela; Nasi, Milena; Gibellini, Lara; DE BIASI, Sara; Benatti, Stefania; Pinti, Marcello; Franceschi, Claudio
abstract

Mitochondrial (mt) DNA or degraded mitochondrial peptides are involved in the pathogenesis of the systemic inflammatory response syndrome (SIRS), a condition that often affects patients who survive a trauma and that resembles sepsis. Indeed, SIRS is due, at least partially, to molecules called "damage-associated molecular patterns" (DAMPs), a family of conserved molecules, conceptually similar to PAMPs, that includes hyaluronan fragments, heat shock proteins, S100 proteins, beta-amyloid, uric acid, IL-1 alpha, IL-33, and the DNA-binding nuclear protein HMGB1. DAMPs can trigger an innate immune response, causing inflammation through the engagement of several TLRs. Mitochondrial DAMPs (MTDs) can be released by damaged tissues, and are highly present in plasma of patients with SIRS. MTDs are represented by formyl peptides, that bind the formyl peptide receptor-1 (FPR-1), and mtDNA, that binds TLR-9, whose activation causes a potent inflammatory reaction.In the last years, we have shown that the production of proinflammatory cytokines is highly increased during the aging process, which is characterized by the accumulation of cellular and molecular defects, and involutive phenomena occurring concomitantly to a hyperstimulation of both innate and adaptive immunity (accumulation of expanded clones of memory and effector T cells, shrinkage of the TCR repertoire, progressive activation of macrophages). These phenomena result in a low-grade, chronic state of inflammation defined as “inflammaging”.To investigate the possible role of extracellular mtDNA during inflammaging, by an original real time PCR assay we have measured plasma levels of mtDNA in individuals who have been followed for 5 years: 429 siblings >90 years old (including 20 ultracentenarians) and 231 controls (50-75 y.o.). Moreover, we have studied 50 young donors (<30 years). A highly significant age-related increase of mtDNA plasma levels was observed. In donors >90 years, mtDNA plasma levels were significantly similar within families, suggesting the existence of a possible genetic component that controls this parameter. mtDNA was not correlated to classical inflammatory markers (CRP, VES), nor to main hematochemical conventional risk factors (among which ALT, albumin, cholesterol and glycemia). Finally, individuals who died within one year from the plasma collection had a lower level of mtDNA, while those who survived >5 years had much higher mtDNA plasma content.We can hypothesize that the age-related increase in plasma mtDNA could be either a marker of an optimal elimination of potentially dangerous cells, or the result of a complex remodeling of the entire organism, that makes use of potentially proinflammatory molecules like MTDs.

2011 - INCREASED LEVELS OF PLASMATIC MITOCHONDRIAL DNA DURING HIV INFECTION REVEAL A NEW ROLE FOR MITOCHONDRIAL DAMAGE-ASSOCIATED MOLECULAR PATTERNS [Abstract in Rivista]
Cossarizza, Andrea; Pinti, Marcello; Nasi, Milena; Gibellini, Lara; DE BIASI, Sara; Montagna, J. P.; Bertoncelli, Linda; Bisi, Luca; Manzini, Lisa; Benatti, Stefania; Borghi, V.; Mussini, Cristina
abstract

Recently, it has been shown that mtDNA or degraded mitochondrial peptides can act as “damage-associated molecular patterns" (DAMPs), that are conceptually and functionally similar to PAMPs. Mitochondrial DAMPs, defined MTDs, can be involved in the pathogenesis of the systemic inflammatory response syndrome (SIRS), a condition that often affects patients who survive a trauma, characterized by the presence of shock and compromised function of several organs. During SIRS, mtDNA released from damaged or dead cells can bind TLR-9, whose activation causes a potent inflammatory reaction, with the production of proinflammatory cytokines. Since HIV infection is characterized by a proinflammatory status, and by a hyperproduction of proinflammatory cytokines, we asked whether soluble mtDNA circulating in the plasma could play a role in determining such condition.Thus, we have studied plasma levels of mtDNA in HIV+ patients showing a different course of the infection, and have correlated such levels to the activation of the immune system and to the plasma viremia. We analyzed individuals during acute, primary HIV infection (AHI), patients with an advanced infection (including those with full blown acquired immunodeficiency syndrome, AIDS) but still naive for antiretroviral therapy (ART), and those defined “long term non progressors” (LTNPs), who had been infected since at least 8 years, always out of treatment, but with a normal number of CD4+ T cells, a low grade of apoptosis and a good immunological control of the virus.In all HIV+ patients but LTNP plasma levels of mtDNA were significantly higher than in healthy controls. Furthermore, in naive patients, 6 months of efficient ART (able to increase CD4+ T cell count, decrease viral load and reduce T cell activation) did not modify mtDNA plasma levels. These levels were not correlated to CD4+ T cell count, nor to markers of immune activation, but had a significant correlation with plasma viral load, revealing a possible role for mtDNA not only as a molecule able to trigger inflammation, but also as a novel biomarker of virus-induced damage.The identification of the role of MTDs could relevant not only to identify possible new biomarkers of disease progression, but also in designing new therapeutic strategies that regard soluble mtDNA, as novel treatments could target either soluble MTDs, or the receptors they use. Thus, in HIV infection, as in other diseases characterized by excessive inflammation, interfering with MTDs could likely become a novel strategy to reduce the harmful immune activation.