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Monica PEDRONI

Personale tecnico amministrativo
Dipartimento di Scienze Mediche e Chirurgiche Materno-Infantili e dell'Adulto

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Pubblicazioni

2022 - Biallelic PMS2 Mutations in a Family with Uncommon Clinical and Molecular Features [Articolo su rivista]
Pedroni, Monica; Ponz de Leon, Maurizio; Reggiani Bonetti, Luca; Rossi, Giuseppina; Viel, Alessandra; Urso, Emanuele Damiano Luca; Roncucci, Luca
abstract

: We describe a patient with constitutional mismatch repair-deficiency (CMMR-D) in whom the syndrome started at age 10 with the development of multiple adenomas in the large bowel. In the successive 25 years, four malignancies developed in different organs (rectum, ileum, duodenum, and lymphoid tissue). The patient had biallelic constitutional pathogenic variants in the PMS2 gene. We speculate that besides the PMS2 genotype, alterations of other genes might have contributed to the development of the complex phenotype. In the nuclear family, both parents carried different PMS2 germline mutations. They appeared in good clinical condition and did not develop polyps or cancer. The index case had a brother who died at age three of lymphoblastic leukemia, and a sister who was affected by sarcoidosis. Tumor tissue showed diffuse DNA microsatellite instability. A complete absence of immunoreactivity was observed for the PMS2 protein both in the tumors and normal tissues. Next-generation sequencing and multiple ligation-dependent probe amplification analyses revealed biallelic PMS2 germline pathogenic variants in the proband (genotype c.[137G>T];[(2174+1_2175-1)_(*160_?)del]), and one of the two variants was present in both parents-c.137G>T in the father and c.(2174+1-2175-1)_(*160_?)del in the mother-as well as c.137G>T in the sister. Moreover, Class 3 variants of MSH2 (c.1787A>G), APC (c.1589T>C), and CHEK2 (c.331G>T) genes were also detected in the proband. In conclusion, the recognition of CMMR-D may sometimes be difficult; however, the possible role of constitutional alterations of other genes in the development of the full-blown phenotype should be investigated in more detail.

2022 - Colon cancer in a 12-year-old girl with hypertriglyceridemia [Articolo su rivista]
Pedroni, Monica; Leon, Maurizio Ponz de; Bonetti, Luca Reggiani; Viel, Alessandra; Noto, Davide; Nascimbeni, Fabio; Sena, Paola; Roncucci, Luca
abstract

2022 - Expression of Autophagic and Inflammatory Markers in Normal Mucosa of Individuals with Colorectal Adenomas: A Cross Sectional Study among Italian Outpatients Undergoing Colonoscopy [Articolo su rivista]
Sena, Paola; Mancini, Stefano; Pedroni, Monica; Reggiani Bonetti, Luca; Carnevale, Gianluca; Roncucci, Luca
abstract

: Colorectal cancer (CRC) ranks among the three most common cancers in terms of both cancer incidence and cancer-related deaths in Western industrialized countries. Lifetime risk of colorectal cancer may reach 6% of the population living in developed countries. In the current era of personalized medicine, CRC is no longer considered as a single entity. In more recent years many studies have described the distinct differences in epidemiology, pathogenesis, genetic and epigenetic alterations, molecular pathways and outcome depending on the anatomical site. The aim of our study is to assess in a multidimensional model the association between metabolic status and inflammatory and autophagic changes in the normal colorectal mucosa classified as right-sided, left-sided and rectum, and the presence of adenomas. One hundred and sixteen patients undergoing colonoscopy were recruited and underwent a complete serum lipid profile, immunofluorescence analysis of colonic biopsies for MAPLC3 and myeloperoxidase expression, matched with clinical and anthropometric characteristics. Presence of adenomas correlated with cholesterol (total and LDL) levels, IL-6 levels, and MAPLC3 tissue expression, especially in the right colon. In conclusion, serum IL-6 amount and autophagic markers could be good predictors of the presence of colorectal adenomas.

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 - Automation of a capture-based NGS workflow: one thousand patients experience in a diagnostic clinical routine framework [Abstract in 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

2019 - Massive juvenile polyposis of the stomach in a family with SMAD4 gene mutation [Articolo su rivista]
de Leon, M. P.; Pedroni, M.; Viel, A.; Luppi, C.; Conigliaro, R.; Domati, F.; Rossi, G.; Bonetti, L. R.
abstract

Relatively little is known on the genotype-phenotype correlations between SMAD4 gene mutations, juvenile polyposis of the intestine and Hereditary Hemorrhagic Teleangectasia. We describe a family in which the proband (a 46-year old woman) had massive polyposis of the stomach—leading to surgery—with high-grade dysplasia at histology. Molecular analysis was carried out using Next Generation sequencing techniques with Miseq Illumina Platforms and a minimal coverage of 40 reads. In the proband, the analysis showed the presence of a truncating mutation in the SMAD4 gene (c.1213dupC, a variant previously associated with juvenile polyposis and Hereditary Hemorrhagic Teleangectasia). The same mutation was detected in two other members of the family (father and brother of the proband), who showed massive polypoid involvement of the stomach at gastroscopy. By taking the family history, subtle evidence of Hereditary Teleangectasia was found (nasal bleeding and arterovenous malformations) in the three gene carriers. Colonoscopy showed polyp occurrence in all three affected members with SMAD4 mutation, with prevalence of adenomatous lesions in one (father), of hamartomas in the brother, and of a mix of histological types in the proband. The main features of the family can be summarized as follows: (A) In hereditary juvenile polyposis, lesions of different histology can be detected at colonoscopy; (B) In the gene carriers of SMAD4 mutations, lesions of the stomach require careful surveillance and, when necessary, surgical interventions; (C) Signs and symptoms of Hereditary Hemorrhagic Teleangectasia should be suspected (and searched) in individuals with SMAD4 constitutional mutations.

2018 - Risk of colorectal polyps and of malignancies in asymptomatic carriers of mutations in the main DNA mismatch repair genes [Articolo su rivista]
Ponz de Leon, Maurizio; Pedroni, Monica; Pezzi, Annalisa; Sulce, Blerta; Roncucci, Luca; Domati, Federica; Rossi, Giuseppina; Reggiani Bonetti, Luca
abstract

Objective: Mutation carriers (Mut+) in DNA mismatch repair genes are predisposed to cancer of various organs and to adenomatous polyps; however, they may remain asymptomatic and cancer or polyp-free for several years. We purposed to analyse the clinical follow-up of individuals carrying constitutional mutations in the MLH1, MSH2 or MSH6 genes who were unaffected by benign polyps or malignant tumours at diagnosis. Material and Methods: Mut + subjects (n.81) were members of Lynch syndromes in whom mutations were detected between 1993 and 2015; all were asymptomatic at diagnosis. They were informed of the cancer risk and surveillance was suggested. As controls, 113 nongene carriers (Mut−) in the same Lynch families were identified. Results: About one-fourth of the mutation carriers developed polyps, mostly adenomas; polyps were less (12%, p <.05) in Mut − subjects, and hyperplastic lesions were the prevalent histology. More polyps were detected in MLH1 vs. MSH2 mutation carriers. In Mut+, 21 malignant tumours developed in 14 carriers vs. 4 tumours in 3 patients among Mut− (p <.001). Tumours were mostly of the Lynch spectrum; however, three glioblastomas were developed, together with neoplasms of various organs (duodenum, thyroid, skin, lung and cervix). Mean age of tumour occurrence was 43.0 years in Mut + vs. 53.0 among Mut−. Conclusions: Cancer developed more often in Mut+, with no consistent difference between MLH1 and MSH2 carriers. More polyps (mostly adenomas) were detected in MLH1 carriers. The majority (13 of 21) of malignant tumours occurred in organs for which there is no recommended surveillance, and were lethal in three patients.

2017 - Attenuated adenomatous polyposis of the large bowel: Present and future [Articolo su rivista]
Roncucci, Luca; Pedroni, Monica; Mariani, Francesco
abstract

Attenuated adenomatous polyposis (AAP) is a poorly understood syndrome, that can be defined as the presence of 10-99 synchronous adenomas in the large bowel, and it is considered a phenotypic variant of familial adenomatous polyposis (FAP). This definition has the advantage of simplicity, but it may include sporadic multiple adenomas of the large bowel at an extreme, or FAP cases on the other side. AAP shows a milder phenotype than FAP, with an older age of onset of adenomas and cancer, and less frequent extracolonic manifestations. AAP may be diagnosed as a single case in a family or, less frequently, it may be present in other family members, and it shows distinct pattern of inheritance. In less than 50% of cases, it may be caused by adenomatous polyposis coli (APC) or MUTYH mutations, referred to as APC-associated polyposis, inherited as an autosomal dominant trait, or MUTYH - associated polyposis, which shows an autosomal recessive mechanism of inheritance, respectively. Surveillance should rely on colonoscopy at regular intervals, with removal of adenomas and careful histological examination. When removal of polyps is not possible or advanced lesions are observed, the surgical approach is mandatory, being subtotal colectomy with ileo-rectal anastomosis the treatment of choice. Studies on this syndrome are lacking, and controversies are still present on many issues, thus, other clinical and genetic studies are requested.

2017 - Attenuated polyposis of the large bowel: a morphologic and molecular approach [Articolo su rivista]
PONZ DE LEON, Maurizio; Pedroni, Monica; Roncucci, Luca; Domati, Federica; Rossi, Giuseppina; Magnani, Giulia; Pezzi, Annalisa; Fante, Rossella; Bonetti, Luca Reggiani
abstract

Attenuated polyposis could be defined as a variant of familial adenomatous polyposis (FAP) in which synchronous polyps of the large bowel range between 10 and 99. We analysed all cases of attenuated polyposis observed over the last 30 years with the objectives: (A) to classify the disease according to different type and proportion of polyps; (B) To ascertain the contribution of APC and MutYH genes; (C) to discover features which could arise the suspicion of mutations; (D) To obtain indications for management and follow-up. 84 individuals in 82 families were studied. Polyps were classified into four groups as adenoma, hyperplastic, other serrated lesions or others; APC and MutYH mutations were assessed. Mean age at diagnosis was 54 ± 14 years in men and 48 ± 13 in women (P = 0.005). Polyps were more numerous in women (37 ± 26 vs 29 ± 22). Sixty % of patients underwent bowel resection, mainly for cancer; the remaining were managed through endoscopy. A total of 2586 polyps were detected at diagnostic endoscopy: 2026 (80 %) were removed and analysed. Adenomas were diagnosed in 1445 (70 %), hyperplastic polyps in 541 (26 %), other serrated lesions in 61 (2.9 %). Adenomas and hyperplastic lesions were detected in the majority of patients. In 68 patients (81 %) in whom studies were executed, APC mutations were found in 8 and MutYH mutations in 10. Genetic variants were more frequent in women (12 vs 6, P = 0.039). Taking into consideration the prevalent (>50 %) histology and presence of mutations, patients could be subdivided into four groups: (1) APC mutated polyposis (AFAP), when adenomas were >50 % and APC mutations detected (no. 8, 10 %); (2) MutYH mutated polyposis (MAP), adenomas >50 % and biallelic MutYH mutations (no. 10, 12 %); (1) attenuated polyposis without detectable mutations, prevalence of adenomas, 48 cases (57 %); (1) hyperplastic-serrated polyposis, with prevalence (>50 %) of hyperplastic/other serrated lesions and no constitutional mutation (no. 18, 21 %). Aggregation of tumors, cancer in probands, distribution of polyps and other clinical characteristics showed no difference among the four groups. In conclusions, AFAP and MAP, the polyposis labeled by constitutional mutations, represented about 25 % of all attenuated polyposis. Mutation-associated cases showed an earlier age of onset of polyps and were more frequent in the female sex.

2017 - Myeloperoxidase-positive cell infiltration of normal colorectal mucosa is related to body fatness and is predictive of adenoma occurrence [Articolo su rivista]
Mariani, F.; Boarino, V.; Bertani, A.; Merighi, A.; Pedroni, M.; Rossi, G.; Mancini, S.; Sena, P.; Benatti, P.; Roncucci, L.
abstract

Body fatness is a risk factor for colorectal cancer, and promotes an inflammatory environment. Indeed, inflammation in normal colorectal mucosa may be a factor linking body fatness to colorectal carcinogenesis. In this study, we evaluated myeloperoxidase (MPO)-positive cells infiltration of normal colorectal mucosa as a marker of cancer-promoting inflammation in overweight and obese subjects. One hundred and three subjects with normal colonoscopy entered the study. Waist circumference (WC) and body mass index (BMI) were measured, and MPO-positive cells on histological sections of biopsies of normal colorectal mucosa were counted under a light microscope. The occurrence of adenomas was then evaluated on follow-up colonoscopies. Mean MPO-positive cell count (±s.e.m.) was higher in subject with a WC equal or above the obesity cutoff values according to gender (2.63±0.20 vs 2.06±0.18, P=0.03), and in subjects with BMI equal or above 25 kg m ' 2 (2.54±0.18 vs 1.97±0.20, P=0.03). A Cox proportional hazard model showed that mean MPO-positive cell count in normal colorectal mucosa was the only factor independently related to occurrence of adenomas in follow-up colonoscopies. Though preliminary, these results show that MPO-positive cell infiltration in normal colorectal mucosa is related with body fatness, as evaluated by WC and BMI, and it may be considered a useful and simple marker to estimate adenoma occurrence risk.

2016 - Prognostic relevance of microsatellite instability in pT3N0M0 colon cancer: a population-based study [Articolo su rivista]
Iachetta, Francesco; Domati, Federica; Reggiani-Bonetti, Luca; Barresi, Valeria; Magnani, Giulia; Marcheselli, Luigi; Cirilli, Claudia; Pedroni, Monica
abstract

Although surgery alone represents a curative approach for patients with pT3N0M0 colon cancer, about 15–20 % of these patients develop a relapse of disease. Microsatellite instability (MSI) is one of the most important molecular markers in colorectal cancer. The aim of this study was to investigate the prognostic relevance of MSI in all pT3N0M0 tumors recorded in the Cancer Registry of the Province of Modena—(Northern Italy) within the 2002–2006 period in patients who showed a relapse of disease during the 5-year period of follow-up (59 cases). They were compared to 59 controls similar in clinical and pathological features but with good prognosis. None of the subjects received adjuvant chemotherapy. MSI status was tested using BAT25, BAT26, NR24, and CAT25 fluorescent-labeled mononucleotide markers. The overall prevalence of MSI was 12.7 % (15 of 118 cases). MSI was detected mainly in mucinous adenocarcinoma (p 

2015 - Aberrant DNA methylation profiles of inherited and sporadic colorectal cancer [Articolo su rivista]
Sahnane, Nora; Magnoli, Francesca; Bernasconi, Barbara; Tibiletti, Maria Grazia; Romualdi, Chiara; Pedroni, Monica; De Leon, Maurizio Ponz; Magnani, Giulia; Reggiani-Bonetti, Luca; Bertario, Lucio; Signoroni, Stefano; Capella, Carlo; Sessa, Fausto; Furlan, Daniela
abstract

Background: Aberrant DNA methylation has been widely investigated in sporadic colorectal carcinomas (CRCs), and extensive work has been performed to characterize different methylation profiles of CRC. Less information is available about the role of epigenetics in hereditary CRC and about the possible clinical use of epigenetic biomarkers in CRC, regardless of the etiopathogenesis. Long interspersed nucleotide element 1 (LINE-1) hypomethylation and gene-specific hypermethylation of 38 promoters were analyzed in multicenter series of 220 CRCs including 71 Lynch (Lynch colorectal cancer with microsatellite instability (LS-MSI)), 23 CRCs of patients under 40 years in which the main inherited CRC syndromes had been excluded (early-onset colorectal cancer with microsatellite stability (EO-MSS)), and 126 sporadic CRCs, comprising 28 cases with microsatellite instability (S-MSI) and 98 that were microsatellite stable (S-MSS). All tumor methylation patterns were integrated with clinicopathological and genetic characteristics, namely chromosomal instability (CIN), TP53 loss, BRAF, and KRAS mutations. Results: LS-MSI mainly showed absence of extensive DNA hypo-and hypermethylation. LINE-1 hypomethylation was observed in a subset of LS-MSI that were associated with the worse prognosis. Genetically, they commonly displayed G:A transition in the KRAS gene and absence of a CIN phenotype and of TP53 loss. S-MSI exhibited a specific epigenetic profile showing low rates of LINE-1 hypomethylation and extensive gene hypermethylation. S-MSI were mainly characterized by MLH1 methylation, BRAF mutation, and absence of a CIN phenotype and of TP53 loss. By contrast, S-MSS showed a high frequency of LINE-1 hypomethylation and of CIN, and they were associated with a worse prognosis. EO-MSS were a genetically and epigenetically heterogeneous group of CRCs. Like LS-MSI, some EO-MSS displayed low rates of DNA hypo-or hypermethylation and frequent G:A transitions in the KRAS gene, suggesting that a genetic syndrome might still be unrevealed in these patients. By contrast, some EO-MSS showed similar features to those observed in S-MSS, such as LINE-1 hypomethylation, CIN, and TP53 deletion. In all four classes, hypermethylation of ESR1, GATA5, and WT1 was very common. Conclusions: Aberrant DNA methylation analysis allows the identification of different subsets of CRCs. This study confirms the potential utility of methylation tests for early detection of CRC and suggests that LINE-1 hypomethylation may be a useful prognostic marker in both sporadic and inherited CRCs.

2015 - Autophagy is upregulated during colorectal carcinogenesis, and in DNA microsatellite stable carcinomas [Articolo su rivista]
Sena, Paola; Mariani, Francesco; Mancini, Stefano; Benincasa, Marta; Magnani, Giulia; Pedroni, Monica; Palumbo, Carla; Roncucci, Luca
abstract

Cancer cells are exposed to a wide range of stress sources, such as nutrient deprivation and hypoxia, as well as cytotoxic chemotherapy and radiotherapy. Certain forms of stress can also promote survival activating the metabolic autophagy pathway in cancer cells. Autophagy is dramatically increased in cancer cells. In these conditions, it is becoming evident that autophagy protects cells, by providing an alternative energy source and by eliminating dysfunctional organelles or proteins. Its role in tumorigenesis is more controversial and both the presence and the absence of autophagy have been implicated. Autophagy is known to be associated with the poor outcome of patients with various types of cancers, and its effectiveness as a prognostic marker in colorectal cancer was demonstrated by several studies. The inhibition of autophagy may be a potential therapeutic target in colorectal cancer. In vitro experiments have shown that the inhibition of autophagy increases 5-FU-induced apoptosis. There are two trials currently investigating the addition of chloroquine to 5-FU-based chemotherapy and bevacizumab. In the present study, we evaluated the expression of LC3B-II in samples of human colorectal microadenomas (i.e., dysplastic aberrant crypt foci) and carcinomas compared to normal mucosa. Furthermore, the expression pattern of LC3B-II was assessed in carcinomas classified as DNA microsatellite stable (MSS) and unstable (MSI). Thus, immunofluorescence techniques coupled with confocal microscopy and immunoblot experiments were performed. The results clearly showed a significant increase in expression of the autophagic key factor in microadenomas and carcinomas with respect to normal mucosa. In MSS carcinomas, the level of LC3B-II expression was higher than that in the MSI carcinomas.

2015 - Molecular features and methylation status in early onset (< 40 years) colorectal cancer: a population based, case-control study [Articolo su rivista]
Magnani, Giulia; Furlan, Daniela; Sahnane, Nora; Reggiani Bonetti, Luca; Domati, Federica; Pedroni, Monica
abstract

Colorectal cancer is usually considered a disease of the elderly. However, a small fraction of patients develops colorectal cancer earlier. The aim of our study was to define the frequency of known hereditary colorectal syndromes and to characterise genetic and epigenetic features of early nonhereditary tumors. Thirty-three patients ≤40 years with diagnosis of colorectal cancer and 41 patients with disease at >60 years of age were investigated for MSI, Mismatch Repair proteins expression, KRAS and BRAF mutations, hypermethylation, and LINE-1 hypomethylation. Detection of germline mutations was performed in Mismatch Repair, APC and MUTYH genes. Early onset colorectal cancer showed a high incidence of hereditary forms (18%). KRAS mutations were detected in 36% of early nonhereditary tumors. Early onset colorectal cancer disclosed an average number of methylated genes significantly lower when compared to the controls (). Finally both of the two groups were highly methylated in ESR1, GATA5, and WT1 genes and were similar for LINE-1 hypomethylation. The genetic make-up of carcinomas differs from young to elderly patients. Early onset tumors showed more frequently a constitutional defective of Mismatch Repair System and a minor number of methylated genes. Hypermethylation of ESR1, GATA5, and WT1 genes suggests possible markers in the earlier diagnosis of colorectal tumorigenesis.

2014 - An unusual case of familial adenomatous polyposis with very early symptom occurrence [Articolo su rivista]
PONZ DE LEON, Maurizio; Bianchini, Maria Anastasia; REGGIANI BONETTI, Luca; Pedroni, Monica; Di Gregorio, Carmela; Merighi, Alberto; Rossi, Giuseppina; Magnani, Giulia; Domati, Federica; Cacciari, Alfredo
abstract

We report the clinical case of a patient who showed an "accelerated" form of polyposis, with development of major lesions within the first decade of life. The patient belongs to a familial adenomatous polyposis family-already described in 2001-featured by profuse polyposis at an early age of onset and desmoid tumors in the majority of affected individuals (of both sexes). The family was characterized by an uncommon mutation of the APC gene (c.4391_4700del310insCACCTACTGCTGAAA, previously defined as c.4394ins15del310) consisting in a large deletion of 310 bp at codon 1,464 with duplication of the breakpoint leading to a stop codon at position 1,575. The proband was affected by desmoids tumors at the age of 3 years. In the same year (2004) numerous polyps in the large bowel and a hepatoblastoma developed. After several months new desmoids appeared in the surgical scar. In 2010, at age 9, the patient was operated of total colectomy and endorectal pull-through of the small intestine owing to profuse colorectal adenomatosis. New desmoids developed in 2011 and 2012, and required chemotherapy. Further analysis of the APC gene in the proband revealed several polymorphisms. One of these (c.398A>G) had not been previously reported, nor was present in two other affected members of the family. The clinical case, and the practical implications for therapy, are discussed according to the most recent theories of colorectal cancer development. Long-term treatment with Cox-2 inhibitors might represent a good option for this patient.

2014 - Application of a 5-tiered scheme for standardized classification of 2,360 unique mismatch repair gene variants in the InSiGHT locus-specific database [Articolo su rivista]
Thompson, Ba; Spurdle, Ab; Plazzer, Jp; Greenblatt, Ms; Akagi, K; Al Mulla, F; Bapat, B; Bernstein, I; Capellá, G; den Dunnen, Jt; du Sart, D; Fabre, A; Farrell, Mp; Farrington, Sm; Frayling, Im; Frebourg, T; Goldgar, De; Heinen, Cd; Holinski Feder, E; Kohonen Corish, M; Robinson, Kl; Leung, Sy; Martins, A; Moller, P; Morak, M; Nystrom, M; Peltomaki, P; Pineda, M; Qi, M; Ramesar, R; Rasmussen, Lj; Royer Pokora, B; Scott, Rj; Sijmons, R; Tavtigian, Sv; Tops, Cm; Weber, T; Wijnen, J; Woods, Mo; Macrae, F; Genuardi, M; Pedroni, Monica
abstract

The clinical classification of hereditary sequence variants identified in disease-related genes directly affects clinical management of patients and their relatives. The International Society for Gastrointestinal Hereditary Tumours (InSiGHT) undertook a collaborative effort to develop, test and apply a standardized classification scheme to constitutional variants in the Lynch syndrome-associated genes MLH1, MSH2, MSH6 and PMS2. Unpublished data submission was encouraged to assist in variant classification and was recognized through microattribution. The scheme was refined by multidisciplinary expert committee review of the clinical and functional data available for variants, applied to 2,360 sequence alterations, and disseminated online. Assessment using validated criteria altered classifications for 66% of 12,006 database entries. Clinical recommendations based on transparent evaluation are now possible for 1,370 variants that were not obviously protein truncating from nomenclature. This large-scale endeavor will facilitate the consistent management of families suspected to have Lynch syndrome and demonstrates the value of multidisciplinary collaboration in the curation and classification of variants in public locus-specific databases.

2014 - Clinical outcome of low- and high-risk malignant colorectal polyps: results of a population-based study and meta-analysis of the available literature [Articolo su rivista]
Di Gregorio, Carmela; Bonetti, Luca Reggiani; DE GAETANI, Carmela; Pedroni, Monica; Kaleci, Shaniko; PONZ DE LEON, Maurizio
abstract

Some histological features of malignant polyps have been used to classify patients into low- and high-risk groups. This study proposed to evaluate the impact of this classification on the clinical outcome of patients with malignant polyps. Through the Colorectal Cancer Registry, 105 patients with endoscopically removed malignant polyps were selected. The presence of one of the following histological features defined malignant polyps as high-risk: infiltrated resection-margin, poorly differentiated carcinoma, lymphatic/vascular invasion and tumour budding and depth of submucosal invasion. Available literature was reviewed by applying a similar classification. Most of the malignant polyps were pedunculated and were localized in the left colon. Fifty-five malignant polyps were classified as low-risk lesions and 50 as high-risk. None of the patients at low-risk died of colorectal cancer. Of the patients at high-risk, three died of cancer; all three cases showed lymphatic/vascular invasion. Review of the literature reveals that an unfavourable clinical outcome is significantly more prevalent in the high-risk compared with the low-risk group (p &gt; 0.005). Moreover, all histological risk factors show a specific predictive value of clinical adverse outcome. Our study and the pooled data analysis confirmed the usefulness of the subdivision into low- and high-risk malignant polyps for management of patients with endoscopically removed colorectal carcinoma.

2014 - Double heterozygosity for BRCA1 and hMLH1 gene mutations in a 46-year-old woman with five primary tumors [Articolo su rivista]
Pedroni, Monica; DI GREGORIO, Carmela; Cortesi, Laura; REGGIANI BONETTI, Luca; Magnani, Giulia; Simone, Maria Luisa; Medici, Veronica; PRIORE OLIVA, Claudio; Marino, Marco; PONZ DE LEON, Maurizio
abstract

Germline mutations in BRCA1 and BRCA2 genes predispose to hereditary breast cancer, whereas carriers of mutations in any of the mismatch repair genes (MMR; hMLH1, hMSH2, hMSH6, hPMS2) are highly susceptible to Lynch syndrome. In the present study, we describe a woman affected by unilateral breast cancer at the age of 35 years. After 4 years, during the follow-up she developed synchronous (and asymptomatic) endometrial cancer, ovarian carcinoma and renal clear cell carcinoma. After 7 years (at age 46), the patient developed an infiltrating carcinoma of the contralateral breast and died in a few months of metastatic disease. Initial investigations led to the detection of a constitutional mutation in the BRCA1 gene. The extended genealogical tree disclosed a suspected history of colorectal carcinoma in the maternal branch. Endometrial cancer of the proband was investigated for microsatellite instability (MSI) and immunohistochemical expression of MLH1, MSH2 and MSH6 proteins. An high MSI status and lack of expression of MLH1 protein were detected. hMLH1 gene sequencing revealed the presence of a constitutional mutation, which was also found in the mother of the proband. Loss of the wild-type hMLH1 allele was detected in both breast tumors, thus suggesting that the MMR defect contributed to the development of the breast cancer.

2014 - Incidence, clinical features and possible etiology of early onset (≤40 years) colorectal neoplasms. [Articolo su rivista]
Domati, Federica; Maffei, Stefania; Kaleci, Shaniko; Di Gregorio, C; Pedroni, Monica; Roncucci, Luca; Benatti, Piero; Magnani, Giulia; Marcheselli, Luigi; Bonetti, Lr; Mariani, Francesco; Alberti, Am; Rossi, V; PONZ DE LEON, Maurizio
abstract

The aim of the study was to investigate the clinical features, including survival, of patients with colorectal malignancies developed at a very early age (≤40 years), together with possible factors involved in the pathogenesis of these rare neoplasms. The study took advantage of the existence of a specialized colorectal cancer Registry active from 1984. 57 patients met the criteria of early onset cancer; main epidemiological data, morphology, stage, familial aggregation, possible role of inheritance and survival were analyzed. Despite the relevant increase over time of all registered patients, joiningpoint analysis of crude incidence rate of early onset colorectal neoplasms revealed a certain stability of these tumors (EAPC: 2.4, CI 14-22) with a constant prevalence of the male sex. Stage at diagnosis did not show significant variations between early onset and maturity onset colorectal neoplasms. Hereditary as well as familial cases were significantly (P &lt; 0.005 and 0.03) more frequent among patients with early onset tumors, although in the majority of them no specific etiological factor could be identified. Survival was more favorable in patients with early onset tumors, though this had to be attributed to the higher presence of some histological types in early onset cases. Survival was significantly more favorable for patients of all ages registered in the last decade. Incidence of early onset colorectal cancer was relatively stable between 1984 and 2008. A male preponderance was evident through the registration period. Hereditary and familial cases were significantly more frequent among early onset case. A well defined etiology could be observed in 16% of the cases (versus 2-3% in older individuals). Five-year survival showed a significant improvement over time.

2014 - Increased expression of autophagy-related proteins in human colorectal cancer development, and correlation with DNA Microsatellite Stable and Unstable [Abstract in Rivista]
Sena, Paola; Mancini, Stefano; Mariani, Francesco; Pedroni, Monica; Magnani, Giulia; Benincasa, Marta; Roncucci, Luca
abstract

Autophagy is upregulated during human colorectal carcinogenesis

2014 - Induction of altered cellular response to oxydative stress in HT29 colon cancer cells treated with metformin [Abstract in Rivista]
Sena, Paola; Mancini, Stefano; Mariani, Francesco; Pedroni, Monica; Benincasa, Marta; Roncucci, Luca
abstract

Metformin induces oxydative stress in HT29 colon cancer cells

2014 - MLH1 constitutional and somatic methylation in patients with MLH1 negative tumors fulfilling the revised Bethesda criteria [Articolo su rivista]
Crucianelli, Francesca; Tricarico, Rossella; Turchetti, Daniela; Gorelli, Greta; Gensini, Francesca; Sestini, Roberta; Giunti, Laura; Pedroni, Monica; PONZ DE LEON, Maurizio; Civitelli, Serenella; Genuardi, Maurizio
abstract

Lynch syndrome (LS) is a tumor predisposing condition caused by constitutional defects in genes coding for components of the mismatch repair (MMR) apparatus. While hypermethylation of the promoter of the MMR gene MLH1 occurs in about 15% of colorectal cancer samples, it has also been observed as a constitutional alteration, in the absence of DNA sequence mutations, in a small number of LS patients. In order to obtain further insights on the phenotypic characteristics of MLH1 epimutation carriers, we investigated the somatic and constitutional MLH1 methylation status of 14 unrelated subjects with a suspicion of LS who were negative for MMR gene constitutional mutations and whose tumors did not express the MLH1 protein. A novel case of constitutional MLH1 epimutation was identified. This patient was affected with multiple primary tumors, including breast cancer, diagnosed starting from the age of 55 y. Investigation of her offspring by allele specific expression revealed that the epimutation was not stable across generations. We also found MLH1 hypermethylation in cancer samples from 4 additional patients who did not have evidence of constitutional defects. These patients had some characteristics of LS, namely early age at onset and/or positive family history, raising the possibility of genetic influences in the establishment of somatic MLH1 methylation.

2014 - MUTYH-associated polyposis (MAP): evidence for the origin of the common European mutations p.Tyr179Cys and p.Gly396Asp by founder events [Articolo su rivista]
Aretz, Stefan; Tricarico, Rossella; Papi, Laura; Spier, Isabel; Pin, Elisa; Horpaopan, Sukanya; Cordisco, Emanuela Lucci; Pedroni, Monica; Stienen, Dietlinde; Gentile, Annamaria; Panza, Anna; Piepoli, Ada; PONZ DE LEON, Maurizio; Friedl, Waltraut; Viel, Alessandra; Genuardi, Maurizio
abstract

MUTYH-associated polyposis (MAP) is an autosomal recessive adenomatous polyposis caused by biallelic germline mutations of the base-excision-repair gene MUTYH. In MAP patients of European origin, the combined allele frequency of the mutations p.Tyr179Cys and p.Gly396Asp ranges between 50 and 82%, while these mutations have not been identified in Far Eastern Asian populations, supporting the hypothesis that a founder effect has occurred at some point in European history. To investigate the natural history of the two common European MUTYH alleles, we genotyped six gene-flanking microsatellite markers in 80 unrelated Italian and German MAP patients segregating one or both mutations and calculated their age in generations (g) by using DMLE+2.2 software. Three distinct common haplotypes, one for p.Tyr179Cys and two for p.Gly396Asp, were identified. Estimated mutation ages were 305 g (95% CS: 271-418) for p.Tyr179Cys and 350 g (95% CS: 313-435) for p.Gly396Asp. These results provide evidence for strong founder effects and suggest that the p.Tyr179Cys and p.Gly396Asp mutations derive from ancestors who lived between 5-8 thousand years and 6-9 thousand years B.C., respectively.

2013 - A case of pneumatosis cystoides intestinalis mimicking familial adenomatous polyposis. [Articolo su rivista]
PONZ DE LEON, Maurizio; Bertarelli, C; Casadei, Gp; Grilli, A; Bacchini, P; Pedroni, Monica; Jovine, E.
abstract

In rare cases the diagnosis of Familial Adenomatous Polyposis (FAP) may not be simple or straightforward. We describe the case of a 54-year man in whom endoscopic and histological features of FAP led to proctocolectomy with ileoanal anastomosis. At anatomical examination, air-filled cystic formations in the submucosa and subserosa of the entire large bowel led to the diagnosis of "Pneumatosis Cystoides Intestinalis", a rare clinical condition featured by the infiltration (or the production) of gas within the intestinal wall. In this case the disease was associated with a long-lasting ingestion of acarbose; as suggested by previous reports, it is possible that gas produced by the fermentation of this unabsorbable carbohydrate penetrates the bowel wall giving origin to cystic formation and to the endoscopic appearance of several polypoid lesions in the large bowel. This report indicates that in rare occasions Pneumatosis Intestinalis can lead to a wrong diagnosis of FAP.

2013 - Incidence trend of malignant polyps through the data of a specialized colorectal cancer registry: clinical features and effect of screening. [Articolo su rivista]
REGGIANI BONETTI, Luca; DI GREGORIO, Carmela; Pedroni, Monica; Domati, Federica; Barresi, V; Marcheselli, L; PONZ DE LEON, Maurizio
abstract

OBJECTIVES: The purposes of the study are to describe the incidence trend of malignant polyp of large bowel over a 25-year period in the District of Modena and to assess the effect of an organized colorectal cancer screening program. MATERIAL AND METHODS: Through the data of a specialized colorectal cancer Registry, we evaluate the clinical and pathological features of the polyps. Trend analysis was assessed with the Joinpoint Regression Program. RESULTS: A total of 172 patients with malignant polyps were diagnosed throughout the study (3.5% of 4.835 registered patients); their overall frequency during the registration period increased from zero cases in the initial years (1984-85) to 57 cases in the past 3 years (2006-2008). Crude incidence rate passed from 0.37 in 1986-89 to 10.2 in 2006. Joinpoint trend analysis of crude rates showed a significant increase of incidence during the study period, with percent of annual variation ranging between 38.6% (95% CI 12.5-70.7) and 7.3% (95% CI 2.6-12.1). During the screening period (2005-2008, the past 4 years of registration) there was a significant increase of sessile polyps (p < 0.001), while other clinical and morphological features, including the number of low- and high-risk malignant polyps, remained unchanged. The surgery (after polypectomy) tended to raise both in low- and high-risk subgroups. CONCLUSION: The incidence of malignant polyps increased significantly from the initial to the most recent periods of colorectal cancer registration. Screening was associated with changes in gross morphology of polyps and with an increased use of the surgery after endoscopic polypectomy.

2013 - Italian cancer figures, report 2013: Multiple tumours [Articolo su rivista]
Adamo, Ms; Alessi, D; Aletta, P; Amodio, R; Andreone, S; Angelin, T; Anghinoni, E; Annulli, Ml; Antonini, S; Artioli, Me; Autelitano, M; Balducci, C; Balottari, P; Baracco, M; Battisti, W; Bella, F; Bellatalla, C; Belluardo, C; Benatti, Piero; Benedetto, G; Benfatto, L; Bernazza, E; Bianconi, F; Biavati, P; Bidoli, E; Birri, S; Bizzoco, S; Bonelli, L; Bonini, A; Borciani, E; Bovo, E; Bozzani, F; Bozzeda, A; Braghiroli, B; Brucculeri, Ma; Brunori, V; Bucalo, G; Bucchi, L; Bugliarello, E; Bulatko, A; Busco, S; Busso, P; Buzzoni, C; Calabretta, L; Caldarella, A; Candela, G; Canu, L; Cappelletti, M; Caprara, L; Carboni, D; Carletti, N; Caroli, S; Carone, S; Cascio, Ma; Cascone, G; Casella, C; Castaing, M; Cecconami, L; Celesia, Mv; Cena, T; Cercato, Mc; Cesaraccio, R; Chiesa, R; Cirilli, C; Civaschi, A; Cocchioni, M; Codazzi, T; Cogno, R; Colamartini, A; Colanino Ziino, A; Cometti, I; Contiero, P; Contrino, Ml; Corbinelli, A; Cordaro, C; Corti, M; Costa, A; Costarelli, D; Cremone, L; Crocetti, E; Curatella, S; Cusimano, R; D'Alò, D; D'Angelo, S; Dal Cappello, T; Dal Cin, A; Dal Maso, L; Dall'Acqua, M; Dalsasso, F; Davini, C; De Dottori, M; De Maria, V; De Santis, E; De Valiere, E; Dei Tos, Ap; Demurtas, G; Devigli, E; Di Felice, E; di Grazia, L; Di Gregorio, C; Di Prima, A; Distefano, R; Doa, N; Domati, F; Fabiano, S; Facchinelli, G; Falcini, F; Falk, M; Fanetti, Ac; Fattoruso, S; Federico, Massimo; Ferrari, L; Ferretti, S; Fidelbo, M; Filipazzi, L; Fiore, Ar; Fiori, G; Foca, F; Forgiarini, O; Frasca, G; Frassoldi, E; Frizza, J; Fusco, M; Fusco, M; Gada, D; Garrone, E; Gasparotti, C; Gatti, L; Gaudiano, C; Gennaro, V; Gentilini, M; Gerevini, C; Ghilardi, S; Ghisleni, S; Giacomin, A; Giavazzi, L; Gilardi, F; Giorgetti, S; Giubelli, C; Giuliani, O; Giurdanella, Mc; Gola, G; Goldoni, Ca; Golizia, Mg; Grandi, L; Greco, A; Guarda, L; Guttadauro, A; Guzzinati, S; Iachetta, F; Iannelli, A; Ieni, A; Intrieri, T; Kaleci, S; La Rosa, F; Lando, C; Lavecchia, Am; Lazzarato, F; Leone, A; Leone, R; Lonati, F; Lottero, B; Lucchi, S; Luminari, Stefano; Macci, L; Macerata, V; Madeddu, A; Maffei, S; Maghini, A; Magnani, C; Magnani, G; Magoni, M; Mameli, G; Mancini, S; Mancuso, P; Mangone, L; Manneschi, G; Mannino, R; Mannino, S; Marani, E; Mariani, F; Martorana, C; Marzola, L; Maspero, S; Maule, M; Mazzei, A; Mazzoleni, G; Mazzucco, G; Melcarne, A; Merletti, F; Michiara, M; Migliari, E; Minerba, S; Minicuzzi, A; Mizzi, M; Monetti, D; Morana, G; Moroni, E; Mosso, Ml; Muni, A; Mura, F; Natali, M; Nemcova, L; Nicita, C; Ocello, C; Paci, E; Pala, F; Palumbo, M; Panico, M; Pannozzo, F; Pascucci, C; Pastore, G; Patriarca, S; Pedroni, Monica; Pellegri, C; Perrotta, C; Pesce, P; Petrinelli, Am; Petrucci, C; Pezzarossi, A; Piffer, S; Pintori, N; Pirani, M; Pirino, D; Pironi, V; PONZ DE LEON, Maurizio; Prandi, R; Prazzoli, R; Preto, L; Puleio, M; Puppo, A; Quaglia, A; Quarta, F; Quattrocchi, M; Raho, Am; Ramazzotti, V; Rashid, I; Ravaioli, A; Ravazzolo, B; Ravegnani, M; Reggiani Bonetti, L; Ribaudo, M; Rinaldi, E; Ricci, P; Rizzello, R; Rollo, Pc; Roncucci, Luca; Rosano, A; Rossi, F; Rossi, G; Rossi, M; Rossini, S; Rosso, S; Rudisi, G; Ruggeri, Mg; Russo, Ag; Russo, M; Sacchettini, C; Sacco, G; Sacerdote, C; Salvatore, S; Salvi, O; Sampietro, G; Sandrini, M; Santucci, C; Scheibel, M; Schiacchitano, S; Sciacca, S; Sciacchitano, C; Scuderi, T; Sechi, O; Seghini, P; Senatore, G; Serafini, G; Serraino, D; Sgargi, P; Sigona, A; Sini, Gm; Sobrato, I; Soddu, M; Solimene, C; Spano, F; Spata, E; Sperduti, I; Staiti, R; Stocco, C; Stracci, F; Sunseri, R; Sardo, As; Tagliabue, G; Tamburo, L; Tamburrino, S; Tanzarella, M; Terracini, B; Tessandori, R; Tisano, F; Tittarelli, A; Tognazzo, S; Torrisi, A; Torrisi, A; Traina, A; Trapani, C; Tschugguel, B; Tumino, R; Usala, M; Vacirca, S; Valerio, O; Valla, K; Varvarà, M; Vasquez, E; Vassante, B; Vattiato, R; Vercelli, M; Vercellino, Pc; Vicentini, M; Villa, M; Vitale, F; Vital
abstract

OBJECTIVES: This collaborative study, based on data collected by the network of Italian association of cancer registries (AIRTUM), provides updated estimates on the incidence risk of multiple primary cancer (MP). The objective is to highlight and quantify the bidirectional associations between different oncological diseases. The quantification of the excess or decreased risk of further cancers in cancer patients, in comparison with the general population, may contribute to understand the aetiology of cancer and to address clinical follow-up. MATERIAL AND METHODS: Data herein presented were provided by AIRTUM population-based cancer registries, which cover nowadays 48% of the Italian population. This monograph utilizes the AIRTUM database (December 2012), considering all malignant cancer cases diagnosed between 1976 and 2010. All cases are coded according to ICD-O-3. Non-melanoma skin cancer cases, cases based on death certificate only, cases based on autopsy only, and cases with follow-up time equal to zero were excluded. To define multiple primaries, IARC-IACR rules were adopted (http://www.iacr.com.fr/MPrules_july2004.pdf). Data were subjected to standard quality control procedures (described in the AIRTUM data management protocol) and specific quality control checks defined for the present study. A cohort of cancer patients was followed over time from first cancer diagnosis until the date of second cancer diagnosis, death, or the end of follow-up, to evaluate whether the number of observed second cancer cases was greater than expected. Person years at risk (PY) were computed by first cancer site, geographic area (North, Centre, South and Islands), attained age, and attained calendar-year group. All second cancers diagnosed in the cohort's patients were included in the observed numbers of cases. The expected number of cancer cases was computed multiplying the accumulated PY by the expected rates, calculated from the AIRTUM database stratified by cancer site, geographic area, age, and calendar-year group. The Standardized Incidence Ratio (SIR) was calculated as the ratio of observed to expected cancer cases. The Excess Absolute Risk (EAR) beyond the expected amount were calculated subtracting the expected number of subsequent cancers from the observed number of cancer cases; the difference was then divided by the PY and the number of cancer cases in excess (or deficit) was expressed per 1,000 PY. Confidence intervals were stated at 95%. The two months (60 days) after first cancer diagnosis were defined as "synchronicity period", and in the main analysis observed and expected cases during this period were excluded. It was estimated the excess risk in the period after first diagnosis (≥ 0 months), excluding the synchronicity period (≥ 2 months), and during the following periods: 2-11, 12-59, 60-119 and 120 months after diagnosis. First-cancer-site-and-gender-specific sheets are presented, reporting both SIRs and EARs. RESULTS: For 5,979,338 person-years a cohort of 1,635,060 cancer patients (880,361 males and 754,699 females) diagnosed between 1976 and 2010 was followed. The mean follow-up length was 14 years. Overall, 85,399 metachronous (latency ≥2 months) cancers were observed, while 77,813 were expected during the study period: SIR: 1.10 (95%CI 1.09-1.10), EAR: 1.32 x 1,000 person-years (95%CI 1.19 - 1.46). The SIR was 1.08 (95%CI 1.08-1.09) for men (54,518 observed and 50,260 expected) and 1.12 (95%CI 1.11-1.13) for women (30,881/27,553), and the EAR 1.61 (95%CI 1.37-1.84) and 1.08 x 1,000 person-years (95%CI 0.93-1.24), respectively.Moreover, during the first two months after first cancer diagnosis (synchronous period) 14,807 cancers were observed while 3,536 were expected (SIR: 4.16; 95%CI 4.09-4.22); the SIR was 4.08 (95%CI 4.00-4.16) for men and 4.32 (95%CI 4.20-4.45) for women.The mean age of patients at first cancer diagnosis was 67.0 years among males and

2013 - Lymph node evaluation in stage IIA colorectal cancer and its impact on patient prognosis: A population-based study [Articolo su rivista]
Iachetta, Francesco; REGGIANI BONETTI, Luca; Marcheselli, Luigi; DI GREGORIO, Carmela; Cirilli, C.; Messinese, S.; Cervo, Gian Luca; Postiglione, Raffaella; DI EMIDIO, Katia; Pedroni, Monica; Longinotti, E.; Federico, Massimo; PONZ DE LEON, Maurizio
abstract

Background. The analysis of regional lymph nodes is particularly relevant in patients with stage II colorectal cancer, in whom the role of adjuvant chemotherapy remains unclear. The aim of this study was to assess the relationship between number of examined lymph nodes and survival in patients with stage IIA (pT3N0M0) colorectal cancer, and to determine the optimal number of lymph nodes that should be examined. Methods. The study group included all the surgically-treated colorectal cancer patients in stage IIA (n = 657) who were identified through the population-based Cancer Registry of the Province of Modena (Northern Italy), during the period 2002-2006. Results. The median number of harvested lymph nodes was 19 (range 1-68). Considering, as a reference point, patients with 12 or less lymph nodes, subjects with n ≥ 20 lymph nodes examined showed, in univariate analysis, a significantly higher cancer specific (p = 0.01) and relapse-free survival (p = 0.003). The results were confirmed by multivariate analysis (Cox model). Conclusion. The result suggests that colorectal cancer patients in stage IIA with n ≥ 20 lymph nodes examined exhibit better survival when compared with subjects in whom fewer lymph nodes were examined. The number of 20 lymph nodes is the essential requirement for an oncologic resection of the large bowel.

2013 - Th Inducing POZ-Kruppel Factor (ThPOK) Is a Key Regulator of the Immune Response since the Early Steps of Colorectal Carcinogenesis [Articolo su rivista]
Mariani, Francesco; Sena, Paola; Pedroni, Monica; Benatti, Piero; Manni, Paola; Di Gregorio, C; Manenti, Antonio; Palumbo, Carla; PONZ DE LEON, Maurizio; Roncucci, Luca
abstract

We purposed to evaluate the role of Th inducing POZ-Kruppel Factor (ThPOK), a transcriptional regulator of T cell fate, in tumour-induced immune system plasticity in colorectal carcinogenesis. The amounts of CD4+, CD8+ and CD56+ and ThPOK+ cells infiltrate in normal colorectal mucosa (NM), in dysplastic aberrant crypt foci (microadenomas, MA), the earliest detectable lesions in colorectal carcinogenesis, and in colorectal carcinomas (CRC), were measured, and the colocalization of ThPOK with the above-mentioned markers of immune cells was evaluated using confocal microscopy. Interestingly, ThPOK showed a prominent increase since MA. A strong colocalization of ThPOK with CD4 both in NM and in MA was observed, weaker in carcinomas. Surprisingly, there was a peak in the colocalization levels of ThPOK with CD8 in MA, which was evident, although to a lesser extent, in carcinomas, too. In conclusion, according to the data of the present study, ThPOK may be considered a central regulator of the earliest events in the immune system during colorectal cancer development, decreasing the immune response against cancer cells.

2012 - Clinical and molecular characterization of colorectal cancer in young Moroccan patients [Articolo su rivista]
Benmoussa, Amal; Badre, Wafaa; Pedroni, Monica; Zamiati, Soumia; Badre, Latifa; Digregorio, Carmela; PONZ DE LEON, Maurizio; Nadifi, Selama
abstract

Early-onset colorectal cancers are relatively rare. About 20% of colorectal cancers are familial or hereditary. Two autosomal dominantly inherited cancer syndromes are more studied: Lynch syndrome accounts for 2-5% of colorectal cancers and familial adenomatous polyposis represents 1% of total colorectal cancers. Unlike the familial adenomatous polyposis syndrome, there are no clinical features that help in easily recognizing Lynch syndrome. The young age of cancer occurrence could be a criterion that should raise a suspicion of Lynch syndrome. In Morocco, the average age at diagnosis of colorectal cancers according to the register of cancers of Casablanca is 56 years, which is 10 years earlier than in European countries. Our study aimed to assess the frequency and molecular characteristics of the Lynch syndrome in Moroccan early-onset colorectal cancers patients.

2012 - Clinical and molecular features of attenuated adenomatous polyposis in northern Italy. [Articolo su rivista]
PONZ DE LEON, Maurizio; Urso, Ed; Pucciarelli, S; Agostini, M; Nitti, D; Roncucci, Luca; Benatti, Piero; Pedroni, Monica; Kaleci, S; Balsamo, A; Laudi, C; Di Gregorio, C; Viel, A; Rossi, G; Venesio, T.
abstract

BACKGROUND: Attenuated familial adenomatous polyposis (AFAP) is characterized by the presence of 10-99 colorectal adenomas. The disease may be associated with mutations in either APC or MUTYH genes. We purposed to evaluate the contribution of adenomatous polyposis coli (APC) and MutY homologue (MUTYH) germline alterations to the AFAP phenotype and to identify genotype/phenotype correlations. METHODS: During counselling for familial adenomatous polyposis (FAP), 91 probands (and 107 affected individuals) who met the criteria of AFAP were identified. Eighty-two families were screened for constitutional mutations of the APC and MUTYH genes. RESULTS: MUTYH mutations were detected in 21 families (25.6 % of the 82 tested), and APC mutations in 7 (8.5 %). Overall, constitutional alterations were found in 34.1 % of the probands. Patients with APC mutations were younger at cancer onset and had a higher mean number of polyps (48.5 ± 33.0 in APC+ individuals vs. 35.7 ± 24.9 in MUTYH+ individuals, and 33.2 ± 18.4 in the "no mutation" group). Clinical features rendered the "no mutation" group closer to MUTYH+ than to the APC+ group. Colorectal cancer at diagnosis was detected in 40 % of AFAP individuals. CONCLUSIONS: AFAP is a new clinical entity with its frequency in the general population still undefined. The number of adenomas varies greatly, with an average of 30-40 lesions. The molecular basis of AFAP can be established in approximately 1/3 of the patients. Both MUTYH and APC genes are implicated in AFAP, though the role of MUTYH is of considerably greater relevance.

2012 - Duodenal carcinoma in a 37-year-old man with Cowden/Bannayan syndrome. [Articolo su rivista]
PONZ DE LEON, Maurizio; Di Gregorio, C; Giunti, L; Roncucci, Luca; Pedroni, Monica; Tinca, Ac; Crucianelli, F; Tricarico, R; Genuardi, M.
abstract

A 37-year-old man was hospitalised because of anaemia and fatigue due to an infiltrating adenocarcinoma of the Treitz angle (duodenum), together with gastric, duodenal and colorectal polyps. After the operation, removal of colorectal lesions revealed the presence of ganglioneuromatosis of the large bowel. Further investigations showed lack of MLH1 protein expression and microsatellite instability in the duodenal neoplasm, while the gene was normally expressed in the polyps. MLH1 sequence and Multiple Ligation-dependent Probes Amplification analysis (from constitutional DNA) were normal. Analysis of the PTEN gene revealed the presence of a constitutional mutation (c.510 T>A; p.Ser170Arg) which had been associated with the Cowden phenotype. Further detailed clinical investigations revealed macrocephaly (63cm), melanotic spots of the penis, small angiomas, millimetric trichilemmomas in the nose and multiple lipomas, which led to the diagnosis of Cowden/Bannayan disease. The unusual appearance of a duodenal carcinoma as the first symptom rendered the identification of the syndrome extremely difficult.

2012 - MSH3 protein expression and nodal status in MLH1-deficient colorectal cancers. [Articolo su rivista]
Laghi, L; Bianchi, P; Delconte, G; Celesti, G; Di Caro, G; Pedroni, Monica; Chiaravalli, Am; Jung, B; Capella, C; PONZ DE LEON, Maurizio; Malesci, A.
abstract

View the MathML source: Colorectal tumors manifesting high-frequency microsatellite instability (MSI-H) develop genetically as a consequence of mutations in genes harboring repetitive DNA sequences. The activin type 2 receptor (ACVR2), possessing 2 polyadenine coding sequences, was identified as a mutational target, but it is not clear if expression is abrogated. Here, we analyzed MSI-H colorectal cancers for ACVR2 mutation and expression to assess if biallelic inactivation occurs. View the MathML source: All 54 MSI-H colon cancers and 20 random microsatellite stable (MSS) tumors from a population-based cohort of 503 patients were analyzed for mutations in 2 A8 tracts (exon 3 and 10) of ACVR2 and the A10 tract of transforming growth factor β receptor 2 (TGFBR2). Additionally, we sequenced exon 10 of ACVR2 in select cancers. ACVR2 expression was determined by immunohistochemistry using an antibody targeting an epitope beyond the predicted truncated protein. View the MathML source: Forty-five of 54 MSI-H cancers (83%) showed mutation (A8 to A7) in the polyadenine tract of exon 10 compared with no MSS tumors. Of tumors with mutant ACVR2, 62% lacked protein expression but all MSS and MSI-H tumors with wild-type ACVR2 expressed protein. We found no evidence of loss of heterozygosity at the ACVR2 locus in MSS tumors. Comparatively, 69% of MSI-H cancers had frameshift mutation in TGFBR2. View the MathML source:ACVR2 mutations are highly frequent in MSI-H colon cancers and in most cases cause loss of ACVR2 expression, indicating biallelic inactivation of the gene. Loss of activin signaling through mutation of ACVR2, similar to observations with TGFBR2, may be important in the genesis of MSI-H colorectal cancer.

2012 - Overweight, inflammation of normal colorectal mucosa, and cancer risk [Abstract in Rivista]
Roncucci, Luca; Mariani, Francesco; Sena, Paola; Palumbo, Carla; Pedroni, Monica
abstract

Myeloperoxidase-positive cell infiltration of normal colorectal mucosa is related to body fatness and colorectal cancer risk.

2011 - Analysis of telomere dynamics in peripheral blood cells from patients with Lynch syndrome. [Articolo su rivista]
Bozzao, C; Lastella, P; PONZ DE LEON, Maurizio; Pedroni, Monica; Di Gregorio, C; D'Ovidio, Fd; Resta, N; Prete, F; Guanti, G; Stella, A.
abstract

In patients with Lynch syndrome, germline mutations in DNA mismatch repair (MMR) genes cause a high risk of developing a broad spectrum of cancers. To date, the management of patients with Lynch syndrome has represented a major challenge because of large variations in age at cancer onset. Several factors, including genetic anticipation, have been proposed to explain this phenotypic heterogeneity, but the molecular mechanisms remain unknown. Telomere shortening is a common event in tumorigenesis and also has been observed in different familial cancers. In this study, the authors investigated the possibility of a relation between telomere length and cancer onset in patients with Lynch syndrome.The mean telomere length was measured using quantitative polymerase chain reaction in peripheral blood samples from a control group of 50 individuals, from 31 unaffected mutation carriers, and from 43 affected patients, and the results were correlated with both gene mutation and cancer occurrence. In affected patients, telomere attrition was correlated with age at cancer onset. In all patients, a t test was used to assess the linearity of the regression.A significant correlation between telomere length and age was observed in both affected and unaffected mutation carriers (P = .0016 and P = .004, respectively) and in mutS homolog 2 (MSH2) mutation carriers (P = .0002) but not in mutL homolog 1 (MLH1) mutation carriers. Telomere attrition was correlated significantly with age at onset in MSH2 carriers (P = .004), whereas an opposite trend toward longer telomeres in patients with delayed onset was observed in MLH1 carriers.The current data suggested that telomere dynamics differ between MLH1 and MSH2 mutation carriers. It is possible that subtle, gene-specific mechanisms can be linked to cancer onset and anticipation in patients with Lynch syndrome.

2011 - I tumori in Italia. Rapporto 2011: La sopravvivenza dei pazienti oncologici in Italia [Articolo su rivista]
Fusco M, AIRTUM Working G. r. o. u. p.; Buzzoni, C; Coviello, E; Rashid, I; Bianconi, F; Cuccaro, F; Castaing, M; De Angelis, R; Giacomin, A; Guzzinati, S; Mosso, Ml; Pisani, P; Quaglia, A; Randi, G; Ramazzotti, V; Russo, A; Senatore, G; Stracci, F; Traina, A; Vercelli, M; Zarcone, M; Ferretti, S; Mazzoleni, G; Bellú, F; Tschugguel, B; De Valiere, E; Facchinelli, G; Falk, M; Dal Cappello, T; Vercellino, Pc; Andreone, S; Busato, A; Marzola, L; Migliari, E; Carletti, N; Nenci, I; Crocetti, E; Caldarella, A; Corbinelli, A; Giusti, F; Intrieri, T; Manneschi, G; Nemcova, L; Romeo, G; Sacchettini, C; Zappa, M; Paci, E; Serraino, D; Angelin, T; Bidoli, E; Dal Maso, L; de Dottori, M; De Paoli, A; De Santis, E; Forgiarini, O; Lise, M; Zucchetto, A; Zanier, L; Orengo, Ma; Casella, C; Marani, E; Puppo, A; Celesia, Mv; Cogno, R; Manenti, S; Garrone, E; Pannozzo, F; Busco, S; Cercato, Mc; Battisti, W; Sperduti, I; Macci, L; Bugliarello, E; Bernazza, E; Tamburo, L; Rossi, M; Curatella, S; De Francesco, C; Tamburrino, S; Bisanti, L; Autelitano, M; Ghilardi, S; Leone, R; Filipazzi, L; Bonini, A; Giubelli, C; Federico, Massimo; Artioli, Me; Valla, K; Braghiroli, B; Cirilli, C; Luminari, Stefano; Pirani, M; Ferrari, L; Bellatalla, C; Fusco, M; Panico, M; Perrotta, C; Vassante, B; Vitale, Mf; Michiara, M; Bozzani, F; Sgargi, P; Tumino, R; La Rosa, Mg; Cascone, G; Frasca, G; Giurdanella, Mc; Martorana, C; Morana, G; Nicita, C; Rollo, Pc; Ruggeri, Mg; Sigona, A; Spata, E; Vacirca, S; Mangone, L; Di Felice, E; Pezzarossi, A; Caroli, S; Pellegri, C; Vicentini, M; Storchi, C; Cavuto, S; Costa, J; Falcini, F; Colamartini, A; Bucchi, L; Balducci, C; Ravegnani, M; Vitali, B; Cordaro, C; Caprara, L; Giuliani, O; Giorgetti, S; Salvatore, S; Palumbo, M; Vattiato, R; Ravaioli, A; Foca, F; Rinaldi, E; Mancini, S; Tonelli, C; Amadori, M; Cremone, L; Iannelli, A; Zevola, A; Budroni, M; Cesaraccio, R; Pirino, D; Carboni, D; Fiori, G; Soddu, M; Mameli, G; Mura, F; Contrino, Ml; Madeddu, A; Tisano, F; Sciacca, S; Muni, A; Mizzi, M; Russo, M; Sacco, G; Aletta, P; Colanino Ziino, A; Tessandori, R; Fanetti, Ac; Maspero, S; Annulli, Ml; Moroni, E; Sanoja Gonzalez, Me; Zanetti, R; Rosso, S; Patriarca, S; Prandi, R; Sobrato, I; Gilardi, F; Busso, P; Piffer, S; Gentilini, Ma; Battisti, L; Rizzello, R; Cappelletti, M; Moser, M; La Rosa, F; D'Alò, D; Scheibel, M; Costarelli, D; Spano, F; Rossini, S; Santucci, C; Petrinelli, Am; Solimene, C; Brunori, V; Crosignani, P; Tagliabue, G; Contiero, P; Preto, L; Tittarelli, A; Maghini, A; Codazzi, T; Frassoldi, E; Gada, D; Costa, E; di Grazia, L; Zambon, P; Baracco, M; Bovo, E; Dal Cin, A; Fiore, Ar; Greco, A; Monetti, D; Rosano, A; Stocco, C; Tognazzo, S; Donato, F; Limina, Rm; Adorni, A; Andreis, P; Zani, G; Piovani, F; Salvi, O; Puleio, M; Vitarelli, S; Antonini, S; Candela, G; Pappalardo, G; Scuderi, T; Lottero, B; Ribaudo, M; Ricci, P; Guarda, L; Gatti, L; Bozzeda, A; Dall'Acqua, M; Pironi, V; Sutera Sardo, A; Mazzei, A; Sirianni, N; Lavecchia, Am; Mancuso, P; Usala, M; Pala, F; Sini, Gm; Pintori, N; Canu, L; Demurtas, G; Doa, N; PONZ DE LEON, Maurizio; Domati, Federica; Rossi, Giuseppina; Goldoni, Ca; Rossi, F; De Gaetani, C; Benatti, Piero; Roncucci, Luca; Di Gregorio, C; Pedroni, Monica; Pezzi, A; Maffei, Stefania; Mariani, Francesco; Borsi, E; Carruba, G; Cusimano, R; Amodio, R; Dolcemascolo, C; Staiti, R; Pastore, G; Magnani, C; Terracini, B; Cena, T; Alessi, D; Baussano, I; Merletti, F; Maule, M; Macerata, V; Cocchioni, M; Pascucci, C; Gennaro, V; Lazzarotto, A; Benfatto, L; Mazzucco, G; Montanaro, F.
abstract

INTRODUCTION: population-based survival analyses are fundamental to assess the impact of public health interventions and new therapies in cancer control. This monograph updates previous reports on cancer patient survival in Italy up to the year 2007. MATERIAL AND METHODS: we extracted from the Network of Italian Cancer Registries (AIRTUM) database over 1,490,000 records of tumours diagnosed during 1990-2007 and followed up to the end of 2008, including all multiple tumours. We used the Ederer II method to estimate relative survival (RS) for 29 different types of neoplasm. Five-year relative survival rates were analysed by gender and macroarea. Trends in 5-, 10- and 15-year RS were studied by gender over six 3-year diagnostic periods, from 1990 to 2007. Conditional 5-year RS was also computed by gender and macroarea. Hybrid approaches were applied to exploit the recent survival experiences of cases diagnosed up to 2007. Adjustment for age was performed using EUROCARE weights. Additional sections describe cancer patient survival in childhood and in elderly patients and provide a comparison of cancer patient survival rates in Italy with those of other countries. RESULTS: Standardized 5-year RS for all tumours but skin in 52% for men and 61% for women. Patient survival has improved for almost all types of cancer: from 1990 to 2007 5-year RS has increased by 15% for all cancers but skin; the exceptions are some cancers with poor prognosis, where patient survival has remained basically unchanged. In males, RS was usually lower than in females, but trend analysis shows that the gap is narrowing. We also report persisting lower RS in southern Italy: 5-year RS in the South is usually from 4% to 10% lower than in the North and Centre. CONCLUSION: this study provides valuable information for all stakeholders in cancer control, both in Italy and elsewhere. Increasing survival reflects improvements in various areas of cancer control. On the other hand, delayed diagnosis and suboptimal management are consistent with the reported differences in survival within the country.

2010 - Analysis of mismatch repair gene mutations in Turkish HNPCC patients. [Articolo su rivista]
Berrin, Tunca; Pedroni, Monica; Gulsah, Cecener; Unal, Egeli; Enrica, Borsi; Abdullah, Zorluoglu; Carmela Di, Gregorio; Tuncay, Yilmazlar; Omaer, Yerci; PONZ DE LEON, Maurizio
abstract

Hereditary non-polyposis colorectal cancer is caused by the inheritance of a mutant allele of a DNA mismatch repair gene. We aimed to investigate types and frequencies of mismatch repair (MMR) gene mutations in Turkish patients with HNPCC and to identify specific biomarkers for early diagnosis of their non-symptomatic kindred’s. The molecular characteristics of 28 Turkish colorectal cancer patients at high-risk for HNPCC were investigated by analysis of microsatellite instability (MSI), immunohistochemistry and methylation-specific PCR in order to select tumors for mutation analysis. The present study adds new information about MMR gene mutation types and their role in Lynch syndrome. This is the first detailed research on Turkish Lynch syndrome families.

2010 - [Italian cancer figures, report 2010: Cancer prevalence in Italy. Patients living with cancer, long-term survivors and cured patients] [Articolo su rivista]
AIRTUM Working, Group; Guzzinati, S.; Dal Maso, L.; De Angelis, R.; De Paoli, A.; Buzzoni, C.; Crocetti, E.; Bucchi, L.; Casella, C.; Cuccaro, F.; Fusco, M.; Luminari, Stefano; Madeddu, A.; Mangone, L.; Patriarca, S.; Piffer, S.; Stracci, F.; Tagliabue, G.; Tumino, R.; Zappa, M.; Capocaccia, R.; Ferretti, S.; Mazzoleni, G.; Bellú, F.; Tschugguel, B.; De Valiere, E.; Facchinelli, G.; Falk, M.; Dal Cappello, T.; Giacomin, A.; Vercellino, P. C.; Andreone, S.; Busato, A.; Marzola, L.; Migliari, E.; Carletti, N.; Nenci, I.; Caldarella, A.; Corbinelli, A.; Giusti, F.; Intrieri, T.; Manneschi, G.; Nemcova, L.; Romeo, G.; Sacchettini, C.; Paci, E.; Serraino, D.; Angelin, T.; Bidoli, E.; de Dottori, M.; De Santis, E.; Forgiarini, O.; Zucchetto, A.; Zanier, L.; Vercelli, M.; Orengo, M. A.; Marani, E.; Puppo, A.; Celesia, M. V.; Cogno, R.; Manenti, S.; Garrone, E.; Quaglia, A.; Pannozzo, F.; Busco, S.; Rashid, I.; Ramazzotti, V.; Cercato, M. C.; Battisti, W.; Sperduti, I.; Macci, L.; Bugliarello, E.; Bernazza, E.; Tamburo, L.; Rossi, M.; Curatella, S.; De Francesco, C.; Tamburrino, S.; Bisanti, L.; Autelitano, M.; Randi, G.; Ghilardi, S.; Leone, R.; Filipazzi, L.; Bonini, A.; Giubelli, C.; Federico, Massimo; Artioli, M. E.; Valla, K.; Braghiroli, B.; Cirilli, C.; Pirani, M.; Ferrari, L.; Bellatalla, C.; Fusco, M.; Panico, M.; Perrotta, C.; Vassante, B.; Traina, A.; Carruba, G.; Cusimano, R.; Amodio, R.; Dolcemascolo, C.; Staiti, R.; Zarcone, M.; Michiara, M.; Bozzani, F.; Sgargi, P.; Cilia, S.; La Rosa, M. G.; Cascone, G.; Frasca, G.; Giurdanella, M. C.; Martorana, C.; Morana, G.; Nicita, C.; Rollo, P.; Ruggeri, M. G.; Sigona, A.; Spata, E.; Vacirca, S.; Di Felice, E.; Pezzarossi, A.; Caroli, S.; Pellegri, C.; Vicentini, M.; Storchi, C.; Cavuto, S.; Costa, J.; Falcini, F.; Colamartini, A.; Balducci, C.; Ravegnani, M.; Vitali, B.; Cordaro, C.; Caprara, L.; Giuliani, O.; Giorgetti, S.; Salvatore, S.; Palumbo, M.; Vattiato, R.; Ravaioli, A.; Foca, F.; Rinaldi, E.; Donato, A.; Iannelli, A.; Senatore, G.; Zevola, A.; Budroni, M.; Cesaraccio, R.; Pirino, D.; Carboni, D.; Fiori, G.; Soddu, M.; Mameli, G.; Mura, F.; Contrino, M. L.; Tisano, F.; Sciacca, S.; Muni, A.; Mizzi, M.; Russo, M.; Tessandori, R.; Ardemagni, G.; Gianola, L.; Maspero, S.; Annulli, M. L.; Moroni, E.; Roberto, G.; Zanetti, R.; Rosso, S.; Prandi, R.; Sobrato, I.; Gilardi, F.; Busso, P.; Franchini, S.; Gentilini, M. A.; Battisti, L.; Cappelletti, M.; Moser, M.; La Rosa, F.; D'Alò, D.; Scheibel, M.; Costarelli, D.; Spano, F.; Rossini, S.; Santucci, C.; Petrinelli, A. M.; Solimene, C.; Bianconi, F.; Brunori, V.; Crosignani, P.; Contiero, P.; Preto, L.; Tittarelli, A.; Maghini, A.; Codazzi, T.; Frassoldi, E.; Gada, D.; Costa, E.; di Grazia, L.; Zambon, P.; Baracco, M.; Bovo, E.; Dal Cin, A.; Fiore, A. R.; Greco, A.; Monetti, D.; Rosano, A.; Stocco, C.; Tognazzo, S.; Donato, F.; Limina, R. M.; Adorni, A.; Andreis, P.; Zani, G.; Piovani, F.; Salvi, O.; Puleio, M.; Vitarelli, S.; Antonini, S.; Candela, G.; Pappalardo, G.; Scuderi, T.; Lottero, B.; Ribaudo, M.; Ricci, P.; Guarda, L.; Gatti, L.; Bozzeda, A.; Dall'Acqua, M.; Pironi, V.; Sutera Sardo, A.; Mazzei, A.; Sirianni, N.; Lavecchia, A. M.; Mancuso, P.; Usala, M.; Pala, F.; Sini, G. M.; Pintori, N.; Canu, L.; Demurtas, G.; Doa, N.; Pisani, P.; Pastore, G.; Magnani, C.; Terracini, B.; Cena, T.; Alessi, D.; Baussano, I.; Merletti, F.; Maule, M.; Mosso, M. L.; Nonnato, M.; Rasulo, A.; Richiardi, L.; Zuccolo, L.; Pivetta, E.; Dalmasso, P.; Macerata, V.; Ponz De Leon, Maurizio; Domati, F.; Rossi, G.; Goldoni, C. A.; Rossi, F.; De Gaetani, C.; Benatti, Piero; Roncucci, Luca; Di Gregorio, C.; Pedroni, Monica; Pezzi, A.; Maffei, S.; Mariani, F.; Borsi, E.; Cocchioni, M.; Pascucci, C.; Gennaro, V.; Lazzarotto, A.; Benfatto, L.; Mazzucco, G.; Montanaro, F.
abstract

OBJECTIVES: the aim of the present monograph is to update the estimation of the number of people living with cancer in Italy, to describe geographic variability, and estimate the number of long-term survivors, i.e., people living five years or more after a cancer diagnosis. MATERIALS AND METHODS: the study included the data of the AIRTUMdatabase. Twenty-four Cancer Registries (CRs) (covering 27% of the Italian population) collected information on the incidence and vital status of 1,275,353 cases diagnosed between 1978 and 2005. For each CR, the observed prevalence was calculated up to the maximum observable duration. To estimate the complete prevalence (all living patients, independently from time since diagnosis) and the prevalence for lengths of time exceeding the CR maximum duration of registration, the observed prevalence was corrected through a completeness index. Completeness indices, gender, age and site specific, were estimated by means of statistical regression models using cancer incidence and survival data available from CRs with more than 15 years of observation. As of 1 January 2006, the prevalence was estimated (as absolute numbers and as a proportion per 100,000 inhabitants) for 46 cancer sites, by gender, age class, years since diagnosis and geographic areas. RESULTS: as of 2006, 2,244,000 persons (4%of the Italian population) were alive with a cancer diagnosis. A relevant geographic variability emerged, with proportions between 4%-5% among CRs in the Centre and North of Italy, and proportions between 2%-3% in the South. Forty-four percent of prevalent subjects (988,000) were males and 56% (1,256,000) females. Fifty-seven percent (1,285,680 people, 2.2% of total population) of these patients was represented by long-term survivors. In patients aged 75 years or more, the proportions of prevalent cases were 19%in males and 13%in females, and 10%between 60 and 75 years of age in both genders.More than half a million Italian women were alive with a breast cancer diagnosis (42%of women with a neoplasm), followed by women with cancers of the colonrectum (12%), corpus uteri (7%), thyroid (5%), and cervix uteri (4%). In men, 22%of prevalent cases (216,716) included patients with prostate cancer, 18% with bladder cancer, and 15%with colon-rectum cancer. Percentages of long-term survivors higher than 70% were reported for cancers of the cervix uteri (82% at five years, and 55% at 15 years from diagnosis), Hodgkin lymphoma, testis, brain and central nervous system, bone and connective tissue. Many patients with these types of cancers (often occurring in young people) can be considered "cured", i.e., with a life expectancy overlapping that of the general population.The estimated proportions of prevalent cases emerging from this study in Italy were quite similar to those reported in Northern Europe, but at least 15%lower than those in the United States. CONCLUSIONS: in 2006, the number of prevalent cases nearly doubled compared to 1992. The increase over time in the proportion of elderly patients, related to population ageing, requires adequate health policies. Knowing the number of people alive many years after cancer diagnosis (either cured or long-term survivors) provides the scientific bases for the definition of health policies focusing on them. Furthermore, it promotes the conduction of studies aimed at improving the present knowledge on the quality of life of these patients during and after the active phase of treatments, in addition to studies on the long-term effects of treatments.

2009 - New incidence and mortality data. 2003-2005 [Articolo su rivista]
Crocetti E, AIRTUM Working G. r. o. u. p.; Buzzoni C., Collaborators:Serraino D; Vicario, G; Angelin, T; Bessega, G; Bidoli, E; Brunetti, D; de Dottori, M; Forgiarini, O; French, S; Stanta, G; Zaina, L; Zanier, L; Zambon, P; Baracco, M; Bovo, E; Dal Cin, A; Fiore, Ar; Greco, A; Guzzinati, S; Monetti, D; Rosano, A; Stocco, Cf; Tognazzo, S; Egarter Vigl, E; Bellù, F; Vittadello, F; Bulatko, A; Lüthy, M; Facchinelli, G; De Valiere, E; Tschugguel, B; Dorfmann, H; Giacomin, A; Vercellino, Pc; Andreone, S; Ferretti, S; Marzola, L; Migliari, E; Carletti, N; Nenci, I; Vitarelli, S; Antonini, S; Federico, Massimo; Artioli, Me; Cirilli, C; Fracca, A; Rashid, I; Valla, K; De Lisi, V; Sgargi, P; Bozzani, F; Donato, A; Iannelli, A; Mari, C; Senatore, G; Zevola, A; Abbamonte, B; Alfano, Ia; Annunziato, L; Barone, S; Ferrante, A; Budroni, M; Cesaraccio, R; Pirino, D; Sechi, O; Piras, D; Sechi, A; Oggiano, M; Piffer, S; Franchini, S; Gentilini, Ma; Battisti, L; Cappelletti, M; Falcini, F; Amadori, D; Balducci, C; Benericetti, E; Bucchi, L; Caprara, L; Colamartini, A; Cordaro, C; Desiderio, F; Fabbri, C; Foca, F; Giorgetti, S; Montanari, E; Naldi, S; Nannini, R; Ravaioli, A; Ravegnani, M; Rinaldi, E; Salvatore, S; Serafini, M; Vattiato, R; Vitali, B; Pannozzo, F; Busco, S; Natali, M; Ramazzotti, V; Macci, L; Bugliarello, E; Bernazza, E; Tamburo, L; Rossi, M; Curatella, S; Sperduti, I; Fusco, M; Bellatalla, C; Fusco, M; Panico, M; Perrotta, C; Vassante, B; Crosignani, P; Tagliabue, G; Contiero, P; Fabiano, S; Maghini, A; Tittarelli, A; Codazzi, T; Frassoldi, E; Costa, E; Nobile, S; Vigano, C; Berrino, F; Mangone, L; Pezzarossi, A; Pellegri, C; Caroli, S; Valentini, M; Cavuto, S; De Felice, E; Vercelli, M; Orengo, Ma; Casella, C; Marani, E; Puppo, A; Celesia, Mv; Cogno, R; Grondona, Am; Giachero, G; Manenti, S; Quaglia, A; Garrone, E; Paci, E; Crocetti, E; Buzzoni, C; Caldarella, A; Corbinelli, A; Dainelli, G; Guadagni, M; Intrieri, T; Manneschi, G; Miccinesi, G; Nemcova, L; Sacchettini, C; Giusti, F; La Rosa, F; Stracci, F; Petrinelli, Am; Costarelli, D; Cassetti, T; Scheibel, M; Romagnoli, C; Mastrandrea, V; Zanetti, R; Rosso, S; Patriarca, S; Vicari, P; Sobrato, I; Gilardi, F; Maglietta, G; Gallesio, L; Tumino, R; Cilia, S; La Rosa, Mg; Cascone, G; Cianciolo, G; Frasca, G; Giurdanella, Mc; Martorana, C; Morana, G; Nicita, C; Rollo, P; Ruggeri, Mg; Sigona, A; Spata, E; Vacirca, S; Bisanti, L; Autelitano, M; Ghilardi, S; Bovini, A; Giubelli, C; Tessandori, R; Ardemagni, G; Traina, A; Candela, P; Contrino, Ml; Tisano, F; Madeddu, A; Ponz de Leon, M; di Gregorio, C; Roncucci, Luca; Benfatti, P; Losi, Lorena; Ponti, Giovanni; Pedroni, Monica; Rossi, G; Roncari, B; Maffei, S; Menigatti, M; Rossi, F; Pecone, L; Domati, F; Pastore, G; Magnani, C; Terracini, B; Alessi, D; Dal masso, P; Dama, E; Macerata, V; Maule, M; Mosso, Ml; Nonnato, M; Zuccolo, L; Merletti, F; Pannelli, F; Pascucci, C; Gennaro, V; Benfatto, L; Bianchelli, M; Lazzarotto, A; Viarengo, P.
abstract

In Italy cancer incidence and mortality rates are similar to those in northern European countries and in USA among males, but they are still lower than women.

2009 - Prognostic relevance of MLH1 and MSH2 mutations in hereditary non-polyposis colorectal cancer patients. [Articolo su rivista]
Russo, A; Sala, P; Alberici, P; Gazzoli, I; Radice, P; Montefusco, C; Torrini, M; Mareni, C; Fornasarig, M; Santarosa, M; Viel, A; Benatti, Piero; Pedroni, Monica; PONZ DE LEON, Maurizio; Lucci Cordisco, E; Genuardi, M; Messerini, L; Stigliano, V; Cama, A; Curia, Mc; de Lellis, L; Signoroni, S; Pierotti, Ma; Bertario, L.
abstract

AIMS AND BACKGROUND: Colorectal carcinoma patients from hereditary non-polyposis colorectal cancer families are suggested to have a better prognosis than sporadic colorectal carcinoma cases. Since the majority of hereditary non-polyposis colorectal cancer-related colorectal carcinomas are characterized by microsatellite instability due to germline mutations in DNA mismatch repair genes, this is consistent with the prolonged survival observed in sporadic microsatellite instability-positive colorectal carcinoma compared to microsatellite stable cases. However, a fraction of colorectal carcinoma cases belongs to families that, despite fulfilling the clinical criteria for hereditary non-polyposis colorectal cancer, do not carry mismatch repair gene mutations. Our aim was to verify to what extent the genotypic heterogeneity influences the prognosis of hereditary non-polyposis colorectal cancer patients. METHODS: A survival analysis was performed on 526 colorectal carcinoma cases from 204 Amsterdam Criteria-positive hereditary non-polyposis colorectal cancer families. Enrolled cases were classified as MLH1-positive, MSH2-positive and mutation-negative, according to the results of genetic testing in each family. RESULTS: Five-year survival rates were 0.73 (95% CI, 0.66-0.80), 0.75 (95% CI, 0.66-0.84) and 0.62 (95% CI, 0.55-0.68) for MLH1-positive, MSH2-positive and mutation-negative groups, respectively (logrank test, P = 0.01). Hazard ratio, computed using Cox regression analysis and adjusted for age, sex, tumor site and stage, was 0.71 (95% CI, 0.51-0.98) for the mutation-positive compared to the mutation-negative group. Moreover, in the latter group, patients with microsatellite instability-positive colorectal carcinomas showed a better outcome than microsatellite stable cases (5-year survival rates, 0.81 and 0.60, respectively; logrank test, P = 0.006). CONCLUSIONS: Our results suggest that the prognosis of hereditary non-polyposis colorectal cancer-related colorectal carcinoma patients depends on the associated constitutional mismatch repair genotype.

2009 - Relative role of APC and MUTYH mutations in the pathogenesis of familial adenomatous polyposis. [Articolo su rivista]
Pezzi, A; Roncucci, Luca; Benatti, Piero; Sassatelli, R; Varesco, L; Di Gregorio, C; Venesio, T; Pedroni, Monica; Maffei, S; Reggiani Bonetti, L; Borsi, E; Ferrari, M; Martella, P; Rossi, G; PONZ DE LEON, Maurizio
abstract

OBJECTIVE: Familial adenomatous polyposis (FAP) is an interesting model for the study of colorectal tumour. Two genes contribute to the FAP phenotype - APC and MUTYH - but their relative role is still undefined. The objective of this study was to evaluate the contribution of the two genes to the pathogenesis of FAP by means of a series of FAP families. MATERIAL AND METHODS: Sixty-one unrelated families with a diagnosis of FAP and a total of 187 affected individuals were evaluated. After extracting DNA, APC and MUTYH genes were sequenced. RESULTS: In the whole series of patients, colectomy with ileorectal anastomosis was the most frequent surgery, although the number of patients treated by total proctocolectomy and ileoanal anastomosis was increasing. Duodenal and jejunal-ileal adenomas were present in more than half of the patients. Constitutional mutations were detected in 37 of the 45 families (82.2%); there were 33 families with APC and 4 with MUTYH alterations. Age at onset of polyposis and age at surgery were 10-15 years delayed for carriers of MUTYH mutations; cancer at diagnosis was frequent, and extracolonic manifestations were diagnosed in the majority of MUTYH-positive families. MUTYH-associated polyposis showed the horizontal transmission expected for recessive inheritance (at variance with the dominant pattern seen with APC mutations). CONCLUSIONS: At least two genes are associated with the FAP phenotype. APC mutations account for the majority of cases, while MUTYH mutations can be observed in 10% of patients. There are few but definite differences between APC- and MUTYH-associated FAP, such as age at diagnosis and pattern of transmission.

2008 - Myeloperoxidase-positive cell infiltration in colorectal carcinogenesis as indicator of colorectal cancer risk. [Articolo su rivista]
Roncucci, Luca; Mora, E; Mariani, F; Bursi, S; Pezzi, A; Rossi, G; Pedroni, Monica; Luppi, D; Santoro, L; Monni, Sebastiano Graziano; Manenti, Antonio; Bertani, A; Merighi, A; Benatti, Piero; Di Gregorio, C; PONZ DE LEON, Maurizio
abstract

Colorectal mucosa is targeted by toxic agents, which can initiate or promote colon cancer. The mechanism of damage might be a focal irritation with loss of normal epithelial cell barrier function. Genetic alterations in tumors may also affect host inflammatory response. The aim of this study was to define the extent of inflammation in colorectal mucosa, along colorectal carcinogenesis, and in microsatellite stable and unstable colorectal carcinomas. We collected 103 samples of normal colorectal mucosa from 65 patients (35 with colorectal cancer or adenoma, 8 with inflammatory bowel diseases, and 22 controls with normal colonoscopy). We also examined 24 aberrant crypt foci, 14 hyperplastic polyps, 16 adenomas, and 67 samples of colorectal carcinoma. Immunohistochemistry was used to count myeloperoxidase (MPO)-positive cells (neutrophils and monocytes) in x100 optical fields under a light microscope. Patients with colorectal tumors had a higher mean number of MPO-positive cells in normal mucosa than controls (mean +/- SD, 2.7 +/- 2.0 versus 1.4 +/- 1.4; P = 0.017). MPO-positive cell number was tightly linked to dysplasia in aberrant crypt foci and adenomas, and it was higher in carcinomas microsatellite unstable than those microsatellite stable (21.6 +/- 15.5 versus 11.9 +/- 8.0; P < 0.01). MPO immunohistochemistry is a simple and reliable technique for the quantification of inflammation in colorectal mucosa., and it may be a potential marker of colorectal cancer risk. Microsatellite instability seems to influence host immune responses to colorectal carcinoma. These observations strongly support a key role of inflammation in colorectal carcinogenesis.

2007 - A mononucleotide markers panel to identify hMLH1/hMSH2 germline mutations. [Articolo su rivista]
Pedroni, Monica; Roncari, B; Maffei, S; Losi, Lorena; Scarselli, A; Di Gregorio, C; Marino, M; Roncucci, Luca; Benatti, Piero; Ponti, Giovanni; Rossi, G; Menigatti, M; Viel, A; Genuardi, M; PONZ DE LEON, Maurizio
abstract

Hereditary NonPolyposis Colorectal Cancer (Lynch syndrome) is an autosomal dominant disease caused by germline mutations in a class of genes deputed to maintain genomic integrity during cell replication, mutations result in a generalized genomic instability, particularly evident at microsatellite loci (Microsatellite Instability, MSI). MSI is present in 85-90% of colorectal cancers that occur in Lynch Syndrome. To standardize the molecular diagnosis of MSI, a panel of 5 microsatellite markers was proposed (known as the "Bethesda panel"). Aim of our study is to evaluate if MSI testing with two mononucleotide markers, such as BAT25 and BAT26, was sufficient to identify patients with hMLH1/hMSH2 germline mutations. We tested 105 tumours for MSI using both the Bethesda markers and the two mononucleotide markers BAT25 and BAT26. Moreover, immunohistochemical evaluation of MLH1 and MSH2 proteins was executed on the tumours with at least one unstable microsatellite, whereas germline hMLH1/hMSH2 mutations were searched for all cases showing two or more unstable microsatellites. The Bethesda panel detected more MSI(+) tumors than the mononucleotide panel (49.5% and 28.6%, respectively). However, the mononucleotide panel was more efficient to detect MSI(+) tumours with lack of expression of Mismatch Repair proteins (93% vs 54%). Germline mutations were detected in almost all patients whose tumours showed MSI and no expression of MLH1/MSH2 proteins. No germline mutations were found in patients with MSI(+) tumour defined only through dinucleotide markers. In conclusion, the proposed mononucleotide markers panel seems to have a higher predictive value to identify hMLH1 and hMSH2 mutation-positive patients with Lynch syndrome. Moreover, this panel showed increased specificity, thus improving the cost/effectiveness ratio of the biomolecular analyses.

2007 - BRAF mutations in multiple sebaceous hyperplasias of patients belonging to MYH-asociated polyposis pedigrees [Articolo su rivista]
Ponti, Giovanni; T., Venesio; Losi, Lorena; Pellacani, Giovanni; L., Bertario; P., Sala; Pedroni, Monica; C., Petti; S., Maffei; L., Varesco; E., Lerch; A., Baggio; Bassoli, Sara; Longo, Caterina; Seidenari, Stefania
abstract

The characteristics of sebaceous gland hyperplasia (SGH) consist of yellowish or skin-colored papules and nodules. Chronic sun exposure and immunosuppressed conditions are the main environmental risk factors, whereas chronological aging regulated by hormones and molecular changes are the intrinsic risk factors. We have evaluated the contribution of BRAF, K-Ras, and N-Ras mutations to the pathogenesis of SGHs in four patients belonging to three MYH-associated polyposis (MAP) pedigrees. MAP is an autosomal-recessive disease characterized by multiple colorectal adenomas and cancer. Immunohistochemistry of mismatch repair and APC proteins was performed. DNA isolated from blood lymphocytes and formalin-fixed or paraffin-em bedded SGHs was PCR amplified and sequenced. In the SGH patients, we detected T1796A heterozygous substitution (V600E) in the BRAF gene. Compound biallelic germline MYH mutations (Y165C/G382D, R168H/379delC, and Y90X/ delGGA464) were detected in the MAP patients. In contrast to the majority of melanocytic lesions, activating hotspot mutations in BRAF have not been involved so far in the pathogenesis of SGH. BRAF mutation is not a specific marker of melanocytic cancerogenesis, and it can also be involved in SGHs. In both melanocytic and non-melanocytic skin tumors, BRAF mutation is linked to early tumorigenesis events.

2007 - Epidemiology of colorectal cancer: the 21-year experience of a specialised registry. [Articolo su rivista]
PONZ DE LEON, Maurizio; Rossi, G; di Gregorio, C; De Gaetani, C; Rossi, F; Ponti, Giovanni; Pecone, L; Pedroni, Monica; Roncucci, Luca; Pezzi, A; Benatti, Piero
abstract

Cancer registries can be viewed as one of the main strategies for improving our understanding of cancer, as they may reveal the importance of specific trends in cancer incidence and survival; in addition, the information obtained from the registries can be translated into preventive measures that might lead to a better control of neoplasms. A colorectal cancer registry was instituted in Northern Italy in 1984. The purpose of this study is to provide a description of the main findings observed in a 21-year period of continuous registration. RESULTS: A total of 3951 malignancies of the large bowel were registered in 3817 patients, for a crude incidence rate of 75.1/100 000/year in men and 59.0 in women. Overall incidence (crude and age-adjusted) of colorectal tumours increased remarkably throughout the registration period. This increase was mainly due to early (Stage I and II) tumours and to lesions with lymph nodal involvement (Stage III). There was a tendency over time towards a progressive increase of colonic tumours, whereas the fraction of rectal neoplasms tended to decline. Colorectal cancer-specific survival increased significantly over time in each of the main TNM/Dukes classes (p<0.006 and <0.001 for Stage II and III tumours). Finally, surgery for colorectal tumours showed a tendency towards large operations (colectomy and hemicolectomy), which was parallel to a definite improvement of pathological staging. CONCLUSIONS: Despite the increasing incidence of colorectal cancer, there are several reasons for cautious optimism. Most of the lesions are now diagnosed at an early stage, and this is associated with a significant increase of survival. The disease is undoubtedly cured better than in the past; the main challenge for future years is to achieve a sustained reduction of mortality for colorectal neoplasms.

2007 - Frequency of constitutional MSH6 mutations in a consecutive series of families with clinical suspicion of HNPCC. [Articolo su rivista]
Roncari, Barbara; Pedroni, Monica; Maffei, S; Di Gregorio, C; Ponti, Giovanni; Scarselli, A; Losi, Lorena; Benatti, Piero; Roncucci, Luca; De Gaetani, C; Camellini, L; Lucci Cordisco, E; Tricarico, E; Genuardi, M; PONZ DE LEON, Maurizio
abstract

A large majority of constitutional mutations in hereditary non-polyposis colorectal cancer (HNPCC) are because of the MHL 1 or MSH 2 genes. In a lower fraction of cases, another gene of the mismatch repair (MMR) machinery, MSH6, may be responsible. Families with MSH6 mutations are difficult to recognize, as microsatellite instability (MSI) may not be detectable and immunohistochemistry (IHC) may give ambiguous results. In the present study, we proposed (i) to determine the frequency of MSH6 mutations in a selected population of colorectal cancer patients obtained from a tumor registry, (ii) to assess whether IHC is a suitable tool for selecting and identifying MSH6 mutation carriers. One hundred neoplasms of the large bowel from suspected HNPCC families were analyzed for MSI (BAT 25 and BAT 26 markers) and immunohistochemical expression of the MSH6 protein. We found on 12 tumors (from different families) showing instability or lack of MSH6 expression. Among these, four potentially pathogenic MSH6 mutations were detected (del A at 2984; del TT at 3119; del AGG cod 385; and del CGT cod 1242) by direct gene sequencing. These represented 12.9% of all families with constitutional mutations of the DNA MMR genes. Thus, some 5% of all HNPCC families are featured by constitutional mutation of the MSH6 gene. This appears, however, as a minimum estimate; routine use of IHC and the study of large numbers of individuals and families with little or no evidence of Lynch syndrome might reveal that mutation of this gene account for a large fraction of HNPCC.

2007 - Genotype-phenotype correlations in individuals with a founder mutation in the MLH1 gene and hereditary non-polyposis colorectal cancer [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, P; Di Gregorio, C; Losi, Lorena; Pedroni, Monica; Ponti, Giovanni; Genuardi, M; Lucci Cordisco, E; Roncucci, Luca
abstract

The results of the study underline the difficulty in discriminating between Lynch I and Lynch II syndromes on the basis of specific molecular changes.

2007 - O6-methylguanine-DNA methyltransferase promoter hypermethylation in colorectal carcinogenesis [Articolo su rivista]
Menigatti, Mirco; Pedroni, Monica; Verrone, Am; Borghi, F; Scarselli, A; Benatti, Piero; Losi, L; Di Gregorio, C; Schär, P; Marra, G; PONZ DE LEON, Maurizio; Roncucci, Luca
abstract

Epigenetic alterations have been reported in colorectal neoplasia which can either complement or in some cases be predisposed to genetic alterations such as K-ras mutations. We examined the promoter methylation status of the CDKN2A and O6-methylguanine-DNA methyltransferase (MGMT) genes, after sodium bisulfite conversion and DNA amplification with methylation specific PCR. Moreover, we searched for G to A transitions in codons 12 and 13 of the K-ras oncogene in normal colorectal mucosae, aberrant crypt foci (ACF, early premalignant lesions) and carcinomas. CDKN2A hypermethylation was an infrequent event in ACF (2 of 26, 7.7%). On the contrary, MGMT hypermethylation was found in the normal mucosae (3 of the 12 samples, 25%), in 14 of the 26 ACF (53.8%) and in 7 of the 9 (77.8%) carcinomas examined. K-ras mutations were evident in 6 ACF (23%) and in 3 carcinomas (33.3%), mostly associated with MGMT promoter hypermethylation. These findings strongly support the hypothesis that epigenetic mechanisms play an important role in the early steps of colorectal carcinogenesis.

2006 - Immunohistochemical expression of MYH protein can be used to identify patients with MYH-associated polyposis [Articolo su rivista]
C., Di Gregorio; M., Frattini; S., Maffei; Ponti, Giovanni; Pedroni, Monica; Venesio, T; Bertario, L; Varesco, L; Risio, M; PONZ DE LEON, Maurizio; Losi, Lorena
abstract

BACKGROUND &amp; AIMS: MYH-associated polyposis is a recently described, autosomal-recessive disease characterized by multiple colorectal adenomas and cancer. There are only few immunohistochemical studies of the MYH protein. We investigated the expression pattern of the MYH protein to evaluate whether a immunohistochemical approach could be used in clinical practice to screen patients for germline mutations in the MYH gene.METHODS: The expression of MYH, MSH2, MLH1, and MSH6 proteins was studied by immunohistochemistry in 20 samples (colorectal adenomas or cancer) from 18 patients with biallelic MYH mutation, in 11 samples from patients with germline adenomatous polyposis coli (APC) mutations, in 20 samples from patients with sporadic colorectal cancers, and in 10 samples from patients with normal colonic mucosa without malignancies.RESULTS: In all cases the mismatch repair proteins were expressed normally. Nuclear and cytoplasmic immunoreactivity for the MYH protein were observed in normal colorectal mucosa, in sporadic colorectal carcinomas, and in adenomas and carcinomas from patients carrying APC germline mutations. Adenomas and carcinomas from patients with MYH biallelic mutation showed a different pattern of expression: a strong granular cytoplasmic staining was observed without any nuclear expression. The same immunophenotype was observed in the surrounding normal mucosa.CONCLUSIONS: Patients with biallelic MYH mutations showed disappearance of staining from the nucleus, and segregation of immunoreactivity in the cytoplasm, both in neoplastic and surrounding healthy mucosa. Because this pattern of expression seems to be specific for biallelic mutations, it follows that immunohistochemistry might be used in clinical practice to screen patients at risk for MYH-associated polyposis.

2006 - Prognostic significance of histological features and biological parameters in stage I (pT1 and pT2) colorectal adenocarcinoma [Articolo su rivista]
Losi, Lorena; Ponti, Giovanni; Di Gregorio, Carmela; Marino, M; Rossi, Giuseppina; Pedroni, Monica; Benatti, Piero; Roncucci, Luca; Ponz De Leon, Maurizio
abstract

Patients with stage I colorectal cancer have a good prognosis, however, a small fraction of them die of local or distant recurrence after curative resection. The aggressive behavior reflects some biological properties of these tumors. In this study, we evaluated the prognostic role of some histopathological and biological parameters in stage I colorectal carcinomas. From the Colorectal Cancer Registry of Modena, we selected two series of patients; the first included all patients who had died of disease progression, the second included patients with a favorable outcome. The histopathological parameters assessed were grade of differentiation, growth pattern at the invasive tumor front, peritumoral lymphocytic infiltration, tumor budding and vascular invasion. The biological variables were proliferative activity (using Ki-67 nuclear antigen), overexpression of p53 protein and altered expression of the mismatch repair proteins (MLH1 and MSH2). The results showed that an infiltrating growth pattern, absent or sparse peritumoral lymphocytic infiltration, the presence of tumor budding and vascular invasion are significantly related to the risk of recurrence. Among the biological parameters, p53 overexpression was significantly correlated with a poor clinical outcome. Our study showed that the histopathologial features are relevant prognostic indicators and might be used as markers for an appropriate treatment strategy in patients with stage I carcinomas.

2006 - Value of MLH1 and MSH2 mutations in the appearance of Muir-Torre syndrome phenotype in HNPCC patients presenting sebaceous gland tumors or Keratoacanthomas [Articolo su rivista]
Ponti, Giovanni; Losi, Lorena; Pedroni, Monica; E., Lucci Cordisco; C., Di Gregorio; Pellacani, Giovanni; Seidenari, Stefania
abstract

Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal-dominant disorder characterized by predisposition to colorectal cancer and extracolonic malignancies, frequent multiple primary tumors in the same patient, and early age of cancer onset. A main clinical variant of Lynch syndrome, Muir-Torre syndrome (MTS) is characterized by the association between one or more visceral malignancies, with at least one sebaceous skin tumor or keratoacanthoma. In our study, we have screened a cohort of 538 HNPCC patients, related to 57 HNPCC families, to detect sebaceous skin tumors and keratoacanthomas and the role of mismatch repair (MMR) genes, MLH1, MSH2, and MSH6, in their pathogenesis. Among the 57 HNPCC families, we have identified four MTS families and one suspected MTS family, in which sebaceous carcinoma was found in one HNPCC mutation carrier subject who did not show visceral malignancy. In four of these families, linked to two MLH1 mutations and to two MSH2 mutations, biomolecular characterization showed concordance among immunohistochemistry analysis and gene mutations. The evidences of our investigations show that MLH1 and MSH2 gene mutations have an equivalent etiopathological role both for Lynch syndrome and for MTS; hence, we propose a broadened clinical criteria for definition of Lynch syndrome that will include sebaceous adenoma, carcinoma, and keratoacanthoma.

2005 - Attenuated familial adenomatous polyposis and Muir-Torre syndrome linked to compound biallelic constitutional MYH gene mutations. [Articolo su rivista]
Ponti, Giovanni; PONZ DE LEON, Maurizio; Maffei, S; Pedroni, Monica; Losi, L; Di Gregorio, C; Gismondi, V; Scarselli, A; Benatti, Piero; Roncari, B; Seidenari, S; Pellacani, Giovanni; Varotti, C; Prete, E; Varesco, L; Roncucci, Luca
abstract

Attenuated familial adenomatous polyposis and Muir-Torre syndrome linked to compound biallelic constitutional MYH gene mutations.Peculiar dermatologic manifestations are present in several heritable gastrointestinal disorders. Muir-Torre syndrome (MTS) is a genodermatosis whose peculiar feature is the presence of sebaceous gland tumors associated with visceral malignancies. We describe one patient in whom multiple sebaceous gland tumors were associated with early onset colon and thyroid cancers and attenuated polyposis coli. Her family history was positive for colonic adenomas. She had a daughter presenting with yellow papules in the forehead region developed in the late infancy. Skin and visceral neoplasms were tested for microsatellite instability and immunohistochemical status of mismatch repair (MMR), APC and MYH proteins. The proband colon and skin tumors were microsatellite stable and showed normal expression of MMR proteins. Cytoplasmic expression of MYH protein was revealed in colonic cancer cells. Compound heterozygosity due to biallelic mutations in MYH, R168H and 379delC, was identified in the proband. The 11-year-old daughter was carrier of the monoallelic constitutional mutation 379delC in the MYH gene; in the sister, the R168H MYH gene mutation was detected. This report presents an interesting case of association between MYH-associated polyposis and sebaceous gland tumors. These findings suggest that patients with MTS phenotype that include colonic polyposis should be screened for MYH gene mutations.

2005 - Different phenotypes in Muir-Torre Syndrome: clinical and biomolecular characterization in two italian families [Articolo su rivista]
Ponti, Giovanni; PONZ DE LEON, Maurizio; Losi, Lorena; C., Di Gregorio; Benatti, Piero; Pedroni, Monica; A., Scarselli; G., Riegler; L., Lembo; Pellacani, Giovanni; Seidenari, Stefania; Rossi, Giorgio; Roncucci, Luca
abstract

The Muir-Torre syndrome (MTS) is an autosomal dominant genodermatosis characterized by the presence of sebaceous gland tumours, with or without keratoacanthomas, associated with visceral malignancies. We describe and characterize two families in which the ample phenotypic variability of MTS was evident. After clinical evaluation, the skin and visceral tumours of one member of a family with 'classic' MTS and one member of a family with a 'peculiar' MTS phenotype without sebaceous lesions, but with only multiple keratoacanthomas, were analysed for microsatellite instability (MSI) and by immunohistochemistry. Tumours of both individuals showed MSI, with a concomitant lack of MSH2 immunostaining in all evaluated skin and visceral lesions; moreover, in the proband of family 2 a constitutional mutation (C -> T substitution leading to a stop codon) in the MSH2 gene was identified. We conclude that the diagnosis of MTS, which is mainly clinical, should take into account an ample phenotypic variability, which includes both cases with typical cancer aggregation in families and cases characterized by the association of visceral malignancies with multiple keratoacanthomas (without sebaceous lesions), without an apparent family history of cancer.

2005 - Identification of Muir-Torre Syndrome among patients with sebaceous tumors and keratoacanthomas: role of clinical features, microsatellite instability and immunohistochemistry [Articolo su rivista]
Ponti, Giovanni; Losi, Lorena; Di Gregorio, C; Roncucci, Luca; Pedroni, Monica; Scarselli, A; Benatti, Piero; Seidenari, S; Pellacani, Giovanni; Lembo, L; Rossi, G; Marino, M; Lucci Cordisco, E; PONZ DE LEON, Maurizio
abstract

BACKGROUND: The Muir-Torre syndrome (MTS) is an autosomal-dominant genodermatosis characterized by the presence of sebaceous gland tumors, with or without keratoacanthomas, associated with visceral malignancies. A subset of patients with MTS is considered a variant of the hereditary nonpolyposis colorectal carcinoma, which is caused by mutations in mismatch-repair genes. The objective of the current study was to evaluate whether a combined clinical, immunohistochemical, and biomolecular approach could be useful for the identification of Muir-Torre syndrome among patients with a diagnosis of sebaceous tumors and keratoacanthomas. METHODS: The authors collected sebaceous skin lesions and keratoacanthomas recorded in the files of the Pathology Department of the University of Modena during the period 1986-2000. Through interviews and examination of clinical charts, family trees were drawn for 120 patients who were affected by these skin lesions. RESULTS: Seven patients also were affected by gastrointestinal tumors, thus meeting the clinical criteria for the diagnosis of MTS. In the MTS families, a wide phenotypic variability was evident, both in the spectrum of visceral tumors and in the type of skin lesions. Microsatellite instability was found in five MTS patients: These patients showed concordance with immunohistochemical analysis; moreover, a constitutional mutation in the MSH2 gene was found in 1 patient. Lack of expression of MSH2/MSH6 or MLH1 proteins was evident in the skin lesions and in the associated internal malignancies of 3 patients and 2 patients with MTS, respectively. CONCLUSIONS: The clinical, biomolecular, and immunohistochemical characterization of sebaceous skin lesions and keratoacanthomas may be used as screening for the identification of families at risk of MTS, a disease that is difficult to recognize and diagnose.

2005 - Incidence and survival of patients with Dukes' A (stages T1 and T2) colorectal carcinoma: a 15-year population-based study. [Articolo su rivista]
Benatti, Piero; Roncucci, Luca; Rossi, G; Ponti, Giovanni; Marino, M; Pedroni, Monica; Scarselli, A; Roncari, Barbara; PONZ DE LEON, Maurizio; Di Gregorio, C; Losi, Lorena
abstract

BACKGROUND AND AIMS: Patients with stage I (Dukes' A) colorectal carcinoma tend to show a good prognosis; however, recurrences can be observed in some patients. Through a specialized colorectal cancer Registry, we attempted to investigate the epidemiological and clinical features of individuals with Dukes' A neoplasms.PATIENTS AND METHODS: From 1984 to 1998, 295 individuals were diagnosed with Stage I /Dukes' A tumors; 150 of these had lesions infiltrating the muscular wall (T2), while 145 had neoplasms limited to the submucosa (T1).RESULTS: Dukes' A tumors represented 13.8% of all registered neoplasms; the percentage doubled over the study period (8.1% in the first year vs. 16.8% in the final year). In each year of observation, the preferential locations were the rectum and sigmoid colon (75% of all lesions). Most patients required surgery, but only 21.3% could be managed by endoscopic polypectomy. Overall 5-year survival was 81.0% (82.1% in T1, 80.0% in T2). Recurrences were seen in 6.8% (2.8% in T1, 10.7% in T2), while 36 patients (12.2%) died of causes unrelated to colorectal cancer. In 17 out of 20 patients who died of cancer, the lesions were localized in the rectosigmoid region. Survival analysis showed a significantly better prognosis (P<0.007) for patients with T1 tumors.CONCLUSIONS: The proportion of stage I colorectal tumors tended to increase over time. Although the overall prognosis is good in four-fifths of the cases, approximately one-fifth of these patients die of recurrent disease or of other causes. As expected, the prognosis was significantly more favorable for patients with T1 lesions. For patients with T2 tumors, radical surgery is the most appropriate approach.

2005 - Investigation of APC mutations in a Turkish familial adenomatous polyposis family by heterodublex analysis [Articolo su rivista]
B., Tunca; M., Menigatti; Benatti, Piero; U., Egeli; G., Cecener; Pedroni, Monica; A., Scarselli; F., Borghi; E., Sala; T., Yilmazlar; A., Zorluoglu; O., Yerci; PONZ DE LEON, Maurizio
abstract

PURPOSE: Familial adenomatous polyposis is an autosomal dominant disease characterized by the presence of 100 or more colorectal adenomatous polyps. Mutations in the adenomatous polyposis coli gene are primarily responsible for the development of this disease. This study was designed to investigation of adenomatous polyposis coli (APC) gene mutations in members of familial adenomatous polyposis family to identify individuals at risk of the disease. METHODS: We examined one patient with familial adenomatous polyposis and 21 family members including one affected person from familial adenomatous polyposis and 20 nonsymptomatic persons. We studied E, D, F, and G segments of exon 15 of the adenomatous polyposis coli gene by heteroduplex analysis. RESULTS: We used silver staining method for staining. We found a mutation for five persons at segment F of exon 15 of the adenomatous polyposis coli gene. Two of them were affected by colorectal cancer, one of whom was the proband, and the other three were non-symptomatic family members. The pathogenetic mutation was a T deletion at codon 1172, causing a frameshift in the adenomatous polyposis coli gene, as a result of the sequencing analysis of these cases. CONCLUSIONS: Investigation of adenomatous polyposis coli gene mutations is very important for the identification of genetic susceptibility to colorectal cancer and for the definition of tumor developing at an early stage. Furthermore, the identification of this mutation for the first time in a Turkish family will be useful to foster further studies on familial adenomatous polyposis in Turkey.

2005 - Microsatellite instability and colorectal cancer prognosis. [Articolo su rivista]
Benatti, Piero; Gafà, R; Barana, D; Marino, M; Scarselli, A; Pedroni, Monica; Maestri, I; Guerzoni, L; Roncucci, Luca; Menigatti, M; Roncari, B; Maffei, S; Rossi, G; Ponti, Giovanni; Santini, A; Losi, Lorena; Di Gregorio, C; Oliani, C; PONZ DE LEON, Maurizio; Lanza, G.
abstract

PURPOSE: Many studies have evaluated the role of high levels of microsatellite instability (MSI) as a prognostic marker and predictor of the response to chemotherapy in colorectal cancer (CRC); however, the results are not conclusive. The aim of this study was to analyze the prognostic significance of high levels of MSI (MSI-H) in CRC patients in relation to fluorouracil-based chemotherapy.EXPERIMENTAL DESIGN: In three different institutions, 1,263 patients with CRC were tested for the presence of MSI, and CRC-specific survival was then analyzed in relation to MSI status, chemotherapy, and other clinical and pathologic variables.RESULTS: Two hundred and fifty-six tumors were MSI-H (20.3%): these were more frequently at a less advanced stage, right-sided, poorly differentiated, with mucinous phenotype, and expansive growth pattern than microsatellite stable carcinomas. Univariate and multivariate analyses of 5-year-specific survival revealed stage, tumor location, grade of differentiation, MSI, gender, and age as significant prognostic factors. The prognostic advantage of MSI tumors was particularly evident in stages II and III in which chemotherapy did not significantly affect the survival of MSI-H patients. Finally, we analyzed survival in MSI-H patients in relation to the presence of mismatch repair gene mutations. MSI-H patients with hereditary non-polyposis colorectal cancer showed a better prognosis as compared with sporadic MSI-H; however, in multivariate analysis, this difference disappeared.CONCLUSIONS: The type of genomic instability could influence the prognosis of CRC, in particular in stages II and III. Fluorouracil-based chemotherapy does not seem to improve survival among MSI-H patients. The survival benefit for patients with hereditary non-polyposis colorectal cancer is mainly determined by younger age and less advanced stage as compared with sporadic MSI-H counterpart.

2005 - Microsatellite instability and prognosis of colorectal cancer [Relazione in Atti di Convegno]
Benatti, Piero; Marino, M; Gafa, R; Barana, D; Pedroni, Monica; Scarselli, A; Di Gregorio, C; Roncucci, Luca; Oliani, C; Lanza, G; PONZ DE LEON, Maurizio
abstract

Purpose: Many studies have evaluated the role of high levels of microsatellite instability (MSI) asa prognostic marker and predictor of the response to chemotherapy in colorectal cancer (CRC);however, the results are not conclusive.The aim of this study was to analyze the prognostic significanceof high levels of MSI (MSI-H) in CRC patients in relation to fluorouracil-based chemotherapy.ExperimentalDesign: In three different institutions,1,263 patientswithCRCwere tested for thepresence of MSI, and CRC-specific survival was then analyzed in relation toMSI status, chemotherapy,and other clinical and pathologic variables.Results:Two hundred and fifty-six tumorswereMSI-H(20.3%): theseweremore frequently at aless advanced stage, right-sided, poorly differentiated, withmucinous phenotype, and expansivegrowth pattern than microsatellite stable carcinomas. Univariate and multivariate analyses of5-year ^ specific survival revealed stage, tumor location, grade of differentiation, MSI, gender,and age as significant prognostic factors.The prognostic advantage of MSI tumors was particularlyevident in stages II and III in which chemotherapy did not significantly affect the survival ofMSI-H patients. Finally, we analyzed survival inMSI-H patients in relation to the presence ofmismatchrepair gene mutations. MSI-H patients with hereditary non ^ polyposis colorectal cancershowed a better prognosis as compared with sporadic MSI-H; however, in multivariate analysis,this difference disappeared.Conclusions:The type of genomic instability could influence the prognosis of CRC, in particularin stages II and III. Fluorouracil-based chemotherapy does not seem to improve survival amongMSI-Hpatients.The survival benefit for patientswithhereditary non ^ polyposis colorectal canceris mainly determined by younger age and less advanced stage as comparedwith sporadicMSI-Hcounterpart.

2005 - Molecular genetic alterations and clinical features in early-onset colorectal carcinomas and their role for the recognition of hereditary cancer syndromes. [Articolo su rivista]
Losi, Lorena; Di Gregorio, C; Pedroni, Monica; Ponti, Giovanni; Roncucci, Luca; Scarselli, A; Genuardi, M; Baglioni, S; Marino, M; Rossi, G; Benatti, Piero; Maffei, S; Menigatti, M; Roncari, Barbara; PONZ DE LEON, Maurizio
abstract

OBJECTIVES: Colorectal cancer (CRC) occurs rarely in young individuals (<45 yr) and represents one of the criteria for suspecting hereditary cancer families. In this study we evaluated clinical features and molecular pathways (chromosomal instability [CIN] and microsatellite instability [MSI]) in early-onset CRC of 71 patients.METHODS: Detailed family and personal history were obtained for each patient. Expression of APC, beta-catenin, p53, MLH1, MSH2, and MSH6 genes was evaluated by immunohistochemistry. MSI analysis was performed and constitutional main mutations of the mismatch repair (MMR) genes were searched by gene sequencing.RESULTS: Fourteen (19.7%) out of the 71 cases showed both MSI and altered expression of MMR proteins. In the 57 MSI-negative (MSI-) lesions altered expression of APC, beta-catenin, and p53 genes were found more frequently than in MSI-positive(MSI+) tumors. Seven (50%) out of the 14 patients with MSI+ tumors presented clinical features of Lynch syndrome (hereditary non-polyposis colorectal cancer [HNPCC]) and in all but one, constitutional mutations in MLH1 or MSH2 genes could be detected. The same mutations were also found in other family members.CONCLUSIONS: Our study demonstrates the involvement of CIN in a majority of early-onset colorectal tumors. Furthermore, we identified Lynch syndromes in seven cases (50%) of early-onset colorectal carcinomas with impairment of the MMR system. These results suggest that patients with early-onset CRC should be screened for hereditary cancer syndrome through clinical and molecular characterizations.

2004 - A founder MLH1 mutation in families from the districts of Modena and Reggio-Emilia in northern Italy with hereditary non-polyposis colorectal cancer associated with protein elongation and instability. [Articolo su rivista]
Caluseriu, O; Lucci Cordisco, E; Santarosa, M; Trojan, J; Brieger, A; Benatti, Piero; Pedroni, Monica; Colibazzi, T; Bellacosa, A; Neri, G; PONZ DE LEON, Maurizio; Viel, A; Genuardi, M.; DI GREGORIO, Carmela
abstract

no abstract

2004 - Aetiology of colorectal cancer and relevance of monogenic inheritance. [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Borghi, Francesca; Pedroni, Monica; Scarselli, Alessandra; DI GREGORIO, C; Losi, Lorena; Viel, A; Genuardi, M; Abbati, G; Rossi, Giuseppina; Menigatti, Mirco; Lamberti, Igor; Ponti, Giovanni; Roncucci, Luca
abstract

BACKGROUND AND AIMS: Although diet and lifestyle are associated with the development of colorectal malignancies, the only clearly identified aetiological factors in colorectal cancer are inheritance (hereditary non-polyposis colorectal cancer (HNPCC) and familial polyposis), inflammatory bowel diseases, papillomavirus, and acquired immunodeficiency syndrome (AIDS). Our aim was to determine what proportion of colorectal neoplasms could be attributed to these specific factors.PATIENTS AND METHODS: Data from a colorectal cancer registry were analysed over a 15 year period, during which nearly 2500 cases were recorded. In patients with suspected HNPCC, microsatellite instability and immunohistochemical expression of proteins encoded by the main DNA mismatch repair genes were assessed. In families with unstable neoplasms, constitutional mutations of the mismatch repair genes hMSH2, hMLH1, and hMSH6 were evaluated by single strand conformation polymorphism analysis and sequencing.RESULTS: Inflammatory bowel diseases, familial polyposis, and AIDS were rare causes of colorectal cancer (three, three, and one case, respectively). Anal squamous carcinoma developed in 27 patients (1.0%) and could be attributed to papillomavirus infection. In 58 patients (from 34 families) a clinical diagnosis of HNPCC was established (2.4%). In total, cases with a known aetiology were 92 (3.7% of all patients). Microsatellite instability was detected in 15 cancers from HNPCC families, and germline mutations in six families (12 patients, 0.5% of the total). Families with unstable tumours, with or without mutations, were clinically similar, suggesting the involvement of the mismatch repair system even when mutations were not detected.CONCLUSIONS: The study suggests that the aetiology of colorectal malignancies remains elusive in the large majority of cases. Among specific causes, HNPCC represents the most frequent. However, with a population based approach, constitutional mutations of the main genes involved in HNPCC can be detected in only 20% of cases.

2004 - Diagnosis of hereditary non-polyposis colorectal cancer (HNPCC) - Reply [Articolo su rivista]
Roncucci, Luca; PONZ DE LEON, Maurizio; Benatti, Piero; Borghi, F; Pedroni, Monica; Scarselli, A; di Gregorio, C; Losi, Lorena; Viel, A; Genuardi, M; Abbati, G; Rossi, G; Menigatti, M; Ponti, Giovanni
abstract

NOTHING

2004 - Diagnosis of hereditary non-polyposis colorectal cancer (HNPCC) [2] (multiple letters) [Articolo su rivista]
Jass, J. R; Roncucci, Luca; PONZ DE LEON, Maurizio; Benatti, Piero; Borghi, F.; Pedroni, Monica; Scarselli, A.; DI GREGORIO, Carmela; Losi, Lorena; Viel, A.; Genuardi, M.; Abbati, Gian Luca; Rossi, G.; Menigatti, Mirco; Ponti, Giovanni
abstract

Letter to the Editor

2004 - Genetic testing among high-risk individuals in families with hereditary non polyposis colorectal cancer [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, P.; Di Gregorio, C.; Pedroni, Monica; Losi, L.; Genuardi, M.; Viel, A.; Fornasarig, M.; Lucci Cordisco, E.; Anti, M.; Ponti, Giovanni; Borghi, F.; Lamberti, I.; Roncucci, Luca
abstract

Hereditary nonpolyposis colorectal cancer (HNPCC) is frequently associated with constitutional mutations in a class of genes involved in DNA mismatch repair. We identified 32 kindreds, with germline mutations in one of three genes hMSH2, hMLH1 or hMSH6. In this study, we purposed to evaluate how many high-risk individuals in each family underwent genetic testing: moreover, we assessed how many mutation-positive unaffected individuals accepted colonoscopic surveillance and the main findings of the recommended follow-up. Families were identified through a population-based registry, or referred from other centres. Members of the families were invited for an education session with two members of the staff. When a kindred was consistent with HNPCC, neoplastic tissues were examined for microsatellite instability (MSI) and immunohistochemical expression of MSH2, MLH1 and MSH6 proteins. Moreover, constitutional mutations were searched by SSCP or direct sequencing of the whole genomic region. Of the 164 subjects assessed by genetic testing, 89 were gene carriers (66 affected - that is, with HNPCC-related cancer diagnosis - and 23 unaffected) and 75 tested negative. Among the 23 unaffected gene carriers, 18 (78.3%) underwent colonoscopy and four declined. On a total of 292 first degree at risk of cancer, 194 (66.4%) did not undergo genetic testing. The main reasons for this were: (a) difficulty to reach family members at risk, (b) lack of collaboration, (c) lack of interest in preventive medicine or 'fatalistic' attitude towards cancer occurrence. The number of colorectal lesions detected at endoscopy in gene carriers was significantly (P&lt;0.01) higher than in controls (noncarriers). We conclude that a large fraction of high-risk individuals in mutation-positive HNPCC families does not undergo genetic testing, despite the benefits of molecular screening and endoscopic surveillance. This clearly indicates that there are still barriers to genetic testing in HNPCC, and that we are unable to provide adequate protection against cancer development in these families.

2004 - Identification of Muir-Torre syndrome among patients with sebaceous tumors and keratoacantomas: role of clinical features, microsatellite instability and immunohistochemistry [Abstract in Rivista]
Losi, Lorena; Ponti, Giovanni; Scarselli, A.; Roncucci, Luca; Pedroni, Monica; P., Benatti; PONZ DE LEON, Maurizio; C., Di Gregorio
abstract

The clinical, biomolecular and immunohistochemical characterization of sebaceous skin lesions and keratoakantomas migh be used in population-based screening for the identification of families at risk of Muir-Torre syndrome, a rare disease difficult to recognize and diagnose.

2004 - Relationship between MUC5AC and altered expression of MLH1 protein in mucinous and non-mucinous colorectal carcinomas [Articolo su rivista]
Losi, Lorena; Scarselli, A; Benatti, Piero; PONZ DE LEON, Maurizio; Roncucci, Luca; Pedroni, Monica; Borghi, F; Lamberti, Igor; Rossi, Giorgio; Marino, M; Ponti, Giovanni; Zangardi, G; Menigatti, M; Di Gregorio, C.
abstract

The main purpose of this study was to examine the expression of mucins and mismatch repair proteins in colorectal carcinomas. The immunohistochemical distribution of apomucins MUC2, MUC5AC, and the expression of MLH1 and MSH2 proteins were examined in 76 mucinous and 60 non-mucinous colorectal carcinomas. MUC2 was noted in all mucinous carcinomas, whereas MUC5AC was present in 41 cases only (54%). In non-mucinous carcinomas, MUC2 was expressed in 61.7% of the tumors; by contrast, MUC5AC was present in 20% of the cases. The expression level of apomucins was significantly different in mucinous and non-mucinous lesions (p < 0.001). Twenty-seven (35.5%) of the mucinous carcinomas showed no MLH1 expression, whereas I I (18.3%) of the non-mucinous tumors did. This difference was statistically significant (p < 0.005). Altered expression of MSH2 protein was never observed. The lack of MLH1 expression was considerably more frequent in carcinomas with secretion of MUC5AC (p<0.005). Our study has demonstrated this close relationship by immunohistochemical methods. In summary, our data show: (1) differences in the expression of mucins between mucinous and non-mucinous tumors; (2) a high frequency of altered MLH1 protein expression (35.5%) in mucinous carcinomas; (3) a significant relationship between the presence of MUC5AC and the altered expression of MLH1 protein in colorectal carcinomas.

2004 - Trend of incidence, subsite distribution and staging of colorectal neoplasms in the 15-year experience of a specialised cancer registry. [Articolo su rivista]
PONZ DE LEON, Maurizio; Marino, M; Benatti, P; Rossi, G; Menigatti, M; Pedroni, Monica; Di Gregorio, C; Losi, Lorena; Borghi, F; Scarselli, A; Ponti, Giovanni; Roncari, B; Zangardi, G; Abbati, G; Ascari, E; Roncucci, Luca
abstract

BACKGROUND: Two-thirds of colorectal malignancies are localised in the left colon and rectum. Recent studies suggest a trend towards an increase of right-sided tumours which might have important implications for screening and surveillance. A colorectal cancer registry was set up in Modena, northern Italy, with the purpose of examining incidence, subsite distribution and staging of colorectal malignancies over a 15-year period.PATIENTS AND METHODS: From 1984 to 1998, 2517 tumours in 2462 patients were detected and staged with the tumour node metastasis (TNM) system. The 'right colon' was considered from caecum to splenic flexure; the 'left colon' included descending and sigmoid colon; and the 'rectum' included rectosigmoid junction, ampulla and anus.RESULTS: Cancer incidence showed an overall increase. Considering the various subsites, an increase of 33.7% in all colonic segments was shown whereas rectal tumours tended to decline. TNM staging showed a gradual increase of localised lesions (41.2% in 1984 versus 53.3% in 1998), with a proportional reduction of advanced tumours.CONCLUSIONS: Our study indicates an increase of tumour incidence in all colonic segments more than a shift to the right colon. TNM staging tended to improve with an appreciable increase of localised lesions. These findings could be consequent to a more extensive use of colonoscopy.

2003 - Caratterizzazione immunoistochimica e biomolecolare del carcinoma colorettale giovanile [Abstract in Rivista]
Losi, Lorena; Di Gregorio, C; Pedroni, Monica; Lamberti, I; Roncucci, Luca; Ponti, Giovanni; Rossi, G; PONZ DE LEON, Maurizio; Benatti, Piero
abstract

Rivista della Società Italiana di Anatomia Patologica e Citopatologia Diagnostica

2003 - Comparison of two marker panels for microsatellite instability analysis in the detection of constitutional MLH1 and MSH2 mutations.. [Abstract in Atti di Convegno]
Roncucci, Luca; Pedroni, Monica; Borghi, F; Scarselli, A; Lamberti, I; Menigatti, M; Rossi, G; Ponti, Giovanni
abstract

Abstract

2003 - Different involvement of target genes mutations in hereditary and sporadic colorectal cancer with Microsatellite Instability. [Poster]
Borghi, F; Pedroni, Monica; Lamberti, I; Menigatti, M; Ponti, Giovanni; Rossi, G; Di Gregorio, C; Losi, Lorena; Scarselli, A; Benatti, P; Roncucci, Luca; PONZ DE LEON, Maurizio
abstract

Functional inactivation of Mismatch Repair (MMR) genes by mutations or epigenetic mechanisms favors the acquisition of mutations in 'target genes'. We analysed 94 colorectal cancer (CRC) with microsatellite instability-high (MSI-H) for the presence of mutations in microsatellites located in the coding regiobns (CDRs) of 6 geens: TGF; RII, BAX, hMLH3, hMSH6, MBD4 and BLM.

2003 - Identification and biomolecular characterization of Muir-Torre Syndrome [Abstract in Rivista]
Ponti, Giovanni; Roncucci, Luca; Di Gregorio, C; Pedroni, Monica; Borghi, F; Lamberti, I; Rossi, G; Abbati, G; Scarselli, A; Riegler, G; Seidenari, S; Pellacani, Giovanni; Lembo, L; Benatti, P; PONZ DE LEON, Maurizio
abstract

Abstract

2003 - L'immunoistochimica delle proteine del mismatch repair può essere un utile test per identificare i pazienti HNPCC? [Abstract in Atti di Convegno]
Di Gregorio, C; Scarselli, A; Pedroni, Monica; Borghi, F; Lamberti, I; Ponti, Giovanni; Roncucci, Luca; Losi, Lorena; PONZ DE LEON, Maurizio; Benatti, P.
abstract

abstract

2003 - Mutations of the hMSH6 geen as a possibel cause of Hereditary Nonpolyposis colorectal cancer [Abstract in Rivista]
PONZ DE LEON, Maurizio; Scarselli, A; Benatti, Piero; Roncucci, Luca; Ponti, Giovanni; Losi, L; Pedroni, Monica; Borghi, F; Menigatti, M; Di Gregorio, C.
abstract

Abstract

2003 - Mutations of the hMSH6 gene as a possible cause of hereditary nonpolyposis colorectal cancer [Abstract in Atti di Convegno]
PONZ DE LEON, Maurizio; Scarselli, A; Benatti, Piero; Roncucci, Luca; Ponti, Giovanni; Losi, Lorena; Pedroni, Monica; Borghi, F; Menigatti, M; Di Gregorio, C.
abstract

Not avaivable

2002 - Alternative marker panel for Microsatellite Instability analysis in deection of contitutional MLH1 and MSH2 mutations. [Abstract in Atti di Convegno]
Pedroni, Monica; Borghi, F; Lamberti, I; Scarselli, A; Menigatti, M; Ponti, Giovanni; Benatti, P; Losi, Lorena; Di Gregorio, C; Abbati, G; Rossi, G; Viel, A; Genuardi, M; Roncucci, Luca; PONZ DE LEON, Maurizio
abstract

Atti del convegno

2002 - Alternative marker panel for microsatellite instability analysis in detection of constitutional MLH1 and MSH2 mutations. [Monografia/Trattato scientifico]
Pedroni, Monica; Borghi, F.; Lamberti, I.; Scarselli, A.; Menigatti, M.; Ponti, Giovanni; Benatti, Piero; Losi, Lorena; Di Gregorio, C.; Abbati, G.; Rossi, Giorgio; Viel, A.; Genuardi, M.; Roncucci, Luca; PONZ DE LEON, Maurizio
abstract

Hereditary Nonpolyposis Colorectal Cancer (I-INPCC) is an autosomal dominant syndrome characterized by predisposition to develop a number of neoplasms including colorectal, endometrium, urinary, extracolonic gastrointestinal, brain and ovarian cancers. HNPCC is caused by inherited mutations in DNA Mismatch Repair (MMR) genes. Defective DNA Mismatch Repair results in genetic instability, which can easily be owerved inshort repetitive sequences as microsatellites (Microsatellite Instability, MSI). An international workshop on MSIheld in Bethesda in 1997 proposed a panel of live microsatellite markers to be used in MSI analysis. This panelincludes two mononucleotide repeats (BAT25, BAT26) and three dinucleotide repeats (D2SI23, D5S346 andDI7S250). In our laboratory we are currently using a di&`erent microsatellite panel composed by threemononucleotide repeats (BAT25, BAT26, BAT40) and two dimmleotide repeats (D2SI23 and Dl8S57). 'I`heaim of this study was to evaluate the specificity ofthe Bethesda markers, as compared with our panel, to idmtifyMLHI and MSIE mutation—positive IINPCC. We compared the results of MSI-analysis in cancer from 27HNPCC Iizmilies (according to the Amsterdam criteria II) and ii•om 75 families in which not all the Amsterdamcriteria were met (Suspected IINPCC), using both the Bethesda and the alternative panel. In addition,immunohistochemistry of MLIII and MSH2 proteins was pertbrmed in all tumors in order to study thecorrelation between the two MSI-panels and the expression of MLIII and MSIE proteins.Using the Bethesda markers, 49 (48%) of tumors showed MSI. On the other hand, using the alternative panel, 33(32,3%) of tumors and displayed MSI. Loss of MLHI or MSIE was evident in 25 of 49 (5I%) MSI tumorsaccording to the Bethesda panel, whereas with the alternative panel 25 of 33 (75,7%) MSI tumors showed noprotein expression. In this group, eleven patients were tested for germline mutations of MMR genes, and all ofthem showed constitutional alterations.Our data suggest that the Bethesda panel is more sensitive to define MSI tumors, but the proposed marker panelis more specific than the Bethesda one to identity MSI tumors with no expression of MLHI and MSIE proteins.'I`he proposed marker panel seems to have a higher predictive value in the identification of

2002 - Biological characterization of mucinous carcinoma of the colon and rectum [Abstract in Rivista]
Losi, Lorena; Scarselli, A; Benatti, P; PONZ DE LEON, Maurizio; Roncucci, Luca; Pedroni, Monica; Borghi, F; Rossi, G; Ponti, Giovanni; Zangardi, G; Menigatti, M; Di Gregorio, C.
abstract

Atti del convegno

2002 - Contributo di un registro tumori sede specifico per il cancro colorettale nell'individuazione di Sindromi rare. [Relazione in Atti di Convegno]
Ponti, Giovanni; Di Gregorio, C; Scarselli, A; Rossi, G; Zangardi, G; Losi, Lorena; Roncucci, Luca; Pedroni, Monica; Borghi, F; Lamberti, I; Benatti, P; PONZ DE LEON, Maurizio
abstract

nd

2002 - HMSH6 immunohistochemistry in patients with clinical suspicion of Hereditary Non-Polyposis Colorectal Cancer. [Monografia/Trattato scientifico]
Scarselli, A.; Benatti, Piero; Chichierchia, G.; Ponti, Giovanni; Lucci Cordisco, E.; Losi, Lorena; Menigatti, M.; Pedroni, Monica; Borghi, F.; Roncucci, Luca; Viel, A.; Genuardi, M.; PONZ DE LEON, Maurizio; Di Gregorio, C.
abstract

Colorectal Cancer (HNPCC) occurs in hMLHl or hMSH2. Recent observations have shown that mutations at hMSl·I6 also involved. Aim of our study is to investigate the role of hMSH6 gene in HNPCC by imnohistochernisny.Materials and methods. 28 colorectal cancer patients with clinical diagnosis of HNPCC or suspected HNPCC were inelected. Immunoliistochemical studies of hMSH6, hMLl—Il and hMSH2 were carried out on paraflin-embedded tumoursamples. lmrm1noperoxidase—staining using diarninobenzidinc as chromogen was carried out with the NEX-ES,Automatic Staining System (Ventana). Mouse monoclonal antibodies to 11MLI-I1 and hMSI-12 proteins (6163-15 andGI29-1129, Pharmingen) were used at 1:40 dilution, mouse monoclonal antibody to hMSH6 protein (clone 44,Transduction Laboratories) was used at 1:2000 dilution. All tumour samples were tested for MSI. Results. Lack of bMSH6 expression was detected in 7 out of 28 tumors. 4 of them also showed absence of hMSH2expression All 7 patients (mean age 56.6 yrs) were affected by right-sided colon cancer, most trequently mucinous and MSI+. 3 patients were from HNPCC families fulfilling Amsterdam Criteria I or I1, 2 were diagnosed as having Muir- Torre syndrome (MTS), and 2 had a diagnosis of suspected HNPCC. An excess of extracolonic tumours was observed in ali the pedigrees but one. interestingly, in both MTS patients, colorectal cancers and sebaceous dermatologic lesionsif thowed the same immunohistochemical pattern. At the moment, the complete MMR gene sequencing has been performed for 3 out of the 7 patients. 2 IJMSH6 and l hMSH2 ramcshifl mutations were detected- Conclusions. l1MSH6 mutations could be characteristic of a subset of HNPCC families. Altered hMSH6 immunohistochemical expression (although often associated with lack of hMSH2 protein), MSI positivity, proximal ` lllcalization, later age at diagnosis and association with extracolonic tumours, all seem to be prognostic features for the presence of this genetic alteration.

2002 - Il registro dei tumori colorettali [Monografia/Trattato scientifico]
PONZ DE LEON, Maurizio; Benatti, Piero; Di Gregorio, C.; Rossi, Giorgio; Losi, Lorena; Foroni, M.; Pedroni, Monica; Menigatti, M.; Zangardi, G.; Roncucci, Luca; Borghi, F.; Scarselli, A.; Percesepe, Antonio; Pasquale, C.
abstract

Not available

2002 - Investigation of APC mutations of a patient with FAP and her family members by heterodublex analyses [Abstract in Atti di Convegno]
Tunca, B; Menigatti, M; Benatti, Piero; Cecener, G; Pedroni, Monica; Scarselli, A; Borghi, F; Sala, E; Yylmazlar, T; Zorluoglu, A; Egeli, U; Yerci, O; PONZ DE LEON, Maurizio
abstract

Familial adenomatous polyposis coli (FAP) is an autosomal dominant disease characterised by the presence of 100 or more colorectal adenomatous polyps. Mutations in the adenomatous polyposis gene(APC) gene primarily responsible for the development of this disease. In this study, we examined one patient with FAP and 21 family members including one effected person from FAP and 20 nonsemptomatic persons. Our proband case who have a retinal lesions (congenital hypertrophy of the retinal pigment epithelium,called CHRPE) and hundreds adenomatous polyps on all colon and rectum is a 36 years old woman. We isolated DNA from pheripheral blood samples of proband and her family members by proteinaz K incubation and phenol-chloroform extraction. We studied E,D, F,and G segments of exon 15 of APC gene by heterodublex analyses (HDA). For staining, we used non-radioactive silver staining method. We determined mutation in 5 person from this family in segmentF of exon 15 of APC. Two of them were patients with FAP (one is ourproband case) and another three persons were non semptomatic family members. Result of sequencing analysis of these cases, wedetermined T deletion at position 3554 causing a frameshift mutation in APC gene.

2002 - Management clinico-endoscopico di soggetti appartenti a famiglie con caratteristiche di sospetta HNPCC:la Sindrome di Muir-Torre. [Abstract in Atti di Convegno]
Ponti, Giovanni; Benatti, Piero; Scarselli, A; Rossi, G; Zangardi, G; Abbati, G; Roncucci, Luca; Pedroni, Monica; Borghi, F; Lamberti, I; Riegler, G; Losi, Lorena; PONZ DE LEON, Maurizio
abstract

Atti del convegno

2002 - Two different marker panels for microsatellie instability analysis in detection of constitutional MLH1 and MSH2 mutations. [Monografia/Trattato scientifico]
Pedroni, Monica; Borghi, F.; Lamberti, I.; Scarselli, A.; Menigatti, M.; Ponti, Giovanni; Benatti, Piero; Losi, Lorena; Di Gregorio, C.; Abbati, G.; Rossi, Giorgio; Roncucci, Luca; PONZ DE LEON, Maurizio
abstract

Hereditary Nonpolyposis Colorectal Cancer (HNPCC), an autosomal dominant susceptibility syndroriie, for approximately 5% of all colorectal tumours. Members of HNPCC families have a predisposition to the of colorectal cancer at an early age and an increased incidence of extracolonic cancer, e.g-, endometrium, ovaries, small bowel, biliary tract, renal pelvis and ureter. Gerrnline mutations in DNA mismatch repair genes, in particular MLI-Il and MSH2, are present in individuals with I-INPCC. Defective DNA mismatch results in genetic instability, which can easily be observed in short repetitive sequences such as microsatellites instability, MSI). MSI has been detected in most MMR-deficient tumours and it is considered the of I-INPCC. An international workshop on MSI held in Bethesda in 1997 proposed a panel of live markers to be used in MSI analysis. This panel, lmown as "Bethesda markers", includes two reats (BAT25, BAT26) and three dinucleotide repeats (D2Sl23, D5S346 and Dl7S250). ln our we are currently using a different microsatellite panel composed by three rnononucleotide repeats (BAT25, BAT40) and two dinucleoide repeats (D2Sl23 and Dl8S57). Aim of our study is to evaluate the speciticity ofr Q Btllhcsda markers, as compared with our panel, to identify MLHI and MSI-I2 mutation-positive HNPCC.We compared the results of MSI~analysis in tumours from 17 HNPCC families (according to and from 75 families in which not all the Amsterdam criteria were met (Suspected HNPCC),both the Bethesda panel and our alternative panel. MSI-H tumours were defined as having instability at two or markers (out of 5), while MSI-L tumours had instability at only one marker and MSS tumours were stable at all In addition, imrrninohistochemistry of MLH1 and MSH2 proteins was performed in all MSI-H and MSI-L to study the correlation between thc two MSI—panels and the expression of MLHI and MSH2 proteins. Results. Using the Bethesda markers, 27 (36%) out of the 75 Suspected HNPCC tumours and I2 (70,6%) out of the HNPCC tumours showed MSI. On the other hand, using our alternative panel, 16 (21.3%) Suspected HNPCC and 7 (41,2%) HNPCC tumours displayed MSI. In Suspected HNPCC tumours we found I5 (20%) md 5 (6.7%) MSI.,using the Bethesda panel and our alternative panel respectively, while in I-INPCC tumours no MSI-L was detected us either panel. Loss of MLHI or MSH2 was evident in 15 out of 39 (38,5%) MSI tumours according to the Beth panel, whereas with our alternative panel 15 out of 23 (65.2%) MSI tumours showed no protein expression.Conclusions. Our data suggest that the Bethesda panel is more sensitive to defne MSI tumours, but the propose marker panel is more sensitive to identify MSI tumours lacking expression of MLHI and MSI-I2 proteins. The marker panel seems to have higher predictive value in the identification of patients with ML!-Il and MSH2 mutations.

2002 - Two different marker panels for microsatellite instability analysis in detection of constitutional MLH1 and MSH2 mutations. [Abstract in Rivista]
Pedroni, Monica; Borghi, F; Lamberti, I; Scarselli, A; Menigatti, M; Ponti, Giovanni; Benatti, P; Losi, L; Di Gregorio, C; Abbati, G; Rossi, G; Roncucci, Luca; PONZ DE LEON, Maurizio
abstract

Hereditary Nonpolyposis Colorectal Cancer (HNPCC), an autosomal dominant susceptibility syndroriie, for approximately 5% of all colorectal tumours. Members of HNPCC families have a predisposition to the of colorectal cancer at an early age and an increased incidence of extracolonic cancer, e.g-, endometrium, ovaries, small bowel, biliary tract, renal pelvis and ureter. Gerrnline mutations in DNA mismatch repair genes, in particular MLI-Il and MSH2, are present in individuals with I-INPCC. Defective DNA mismatch results in genetic instability, which can easily be observed in short repetitive sequences such as microsatellites instability, MSI). MSI has been detected in most MMR-deficient tumours and it is considered the of I-INPCC. An international workshop on MSI held in Bethesda in 1997 proposed a panel of live markers to be used in MSI analysis. This panel, lmown as "Bethesda markers", includes two reats (BAT25, BAT26) and three dinucleotide repeats (D2Sl23, D5S346 and Dl7S250). ln our we are currently using a different microsatellite panel composed by three rnononucleotide repeats (BAT25, BAT40) and two dinucleoide repeats (D2Sl23 and Dl8S57). Aim of our study is to evaluate the speciticity of r Q Btllhcsda markers, as compared with our panel, to identify MLHI and MSI-I2 mutation-positive HNPCC.We compared the results of MSI~analysis in tumours from 17 HNPCC families (according to and from 75 families in which not all the Amsterdam criteria were met (Suspected HNPCC), both the Bethesda panel and our alternative panel. MSI-H tumours were defined as having instability at two or markers (out of 5), while MSI-L tumours had instability at only one marker and MSS tumours were stable at all In addition, imrrninohistochemistry of MLH1 and MSH2 proteins was performed in all MSI-H and MSI-L to study the correlation between thc two MSI—panels and the expression of MLHI and MSH2 proteins. Results. Using the Bethesda markers, 27 (36%) out of the 75 Suspected HNPCC tumours and I2 (70,6%) out of the HNPCC tumours showed MSI. On the other hand, using our alternative panel, 16 (21.3%) Suspected HNPCC and 7 (41,2%) HNPCC tumours displayed MSI. In Suspected HNPCC tumours we found I5 (20%) md 5 (6.7%) MSI., using the Bethesda panel and our alternative panel respectively, while in I-INPCC tumours no MSI-L was detected us either panel. Loss of MLHI or MSH2 was evident in 15 out of 39 (38,5%) MSI tumours according to the Beth panel, whereas with our alternative panel 15 out of 23 (65.2%) MSI tumours showed no protein expression.Conclusions. Our data suggest that the Bethesda panel is more sensitive to defne MSI tumours, but the propose marker panel is more sensitive to identify MSI tumours lacking expression of MLHI and MSI-I2 proteins. The marker panel seems to have higher predictive value in the identification of patients with ML!-Il and MSH2 mutations.

2002 - hMSH6 immunohistochemistry in patients with clinical suspicion of Hereditary Non-Polyposis Colorectal Cancer. [Abstract in Atti di Convegno]
Scarselli, A; Benatti, Piero; Chichierchia, G; Ponti, Giovanni; Lucci Cordisco, E; Losi, Lorena; Menigatti, M; Pedroni, Monica; Borghi, F; Roncucci, Luca; Viel, A; Genuardi, M; PONZ DE LEON, Maurizio; Di Gregorio, C.
abstract

Congresso Italiano Patologia e Diagnostica Molecolare

2001 - Clinical and biologic heterogeneity of Hereditary NonPolyposis Colorectal Cancer. [Articolo su rivista]
Benatti, Piero; Roncucci, Luca; Ganazzi, Dorval; Percesepe, Antonio; Di Gregorio, C.; Pedroni, Monica; Borghi, F.; Sala, E.; Scarselli, A.; Menigatti, M.; Rossi, Giuseppina; Genuardi, M.; Viel, A.; PONZ DE LEON, Maurizio
abstract

MMR gene mutations and MSI are not found in all clinically diagnosed HNPCC families. We evaluated whether MMR genotyping and tumor MSI analysis could identify distinct clinical subgroups among HNPCC families. Twenty-nine clinical HNPCC families were divided into 3 groups: A, families with hMLH1 or hMSH2 gene mutations; B, MMR gene mutations not present but MSI present in at least 50% of tumors tested; C, mutational and MSI analyses negative. We evaluated tumor spectrum, age at onset, risk of cancer in the follow-up and survival for CRC in the 3 groups. Tumors of the target organs in HNPCC (colon and rectum, endometrium, ovary, small bowel, stomach, renal pelvis and ureter) were more frequent in the first 2 groups than in the latter. Colon cancer was more frequently located in the proximal colon and showed an earlier age at onset in families with MMR gene mutation or with MSI than in families with stable tumors. Comparing the occurrence of tumors in the follow-up, in the first 2 groups patients younger than 50 years had a higher RR, which was particularly marked for CRC (RR = 18.6 for group A vs. group C, RR = 16.7 for group B vs. group C). CRC patients in the first 2 groups had a better clinical prognosis. The results of molecular analysis could distinguish, within clinically defined HNPCC families, different subgroups to which specific programs of surveillance could be addressed. Copyright 2001 Wiley-Liss, Inc.

2001 - Epidemiologia dei tumori del colon-retto. Incidenza, mortalità, familiarità e sopravvivenza nella ex USL di Modena, 1984-1998. [Monografia/Trattato scientifico]
PONZ DE LEON, Maurizio; Benatti, Piero; Rossi, Giorgio; Di Gregorio, C.; Roncucci, Luca; Losi, Lorena; Foroni, M.; Pedroni, Monica; Menigatti, M.; Zangardi, G.; Scarselli, A.; Percesepe, Antonio; Borghi, F.; Pasquale, C.
abstract

not available

2001 - Genotype-phenotype association in Italian HNPCC families:preliminary results and proposal of a National collaborative study. [Monografia/Trattato scientifico]
Benatti, Piero; Lucci Cordisco, E.; Caluseriu, O.; Pedroni, Monica; Scarselli, A.; Losi, Lorena; Menigatti, M.; Borghi, F.; Roncucci, Luca; Viel, A.; Genuardi, M.; PONZ DE LEON, Maurizio
abstract

not available

2001 - Methylation pattern of different regions of the MLH1 promoter and silencing of gene expression in hereditary and sporadic colorectal cancer. [Articolo su rivista]
Di Gregorio, C; Borghi, F; Sala, E; Scarselli, A; Pedroni, Monica; Foroni, M; Benatti, Piero; Roncucci, Luca; PONZ DE LEON, Maurizio; Percesepe, Antonio; Menigatti, M
abstract

Nonrandom, widespread promoter methylation of tumor suppressor genes is a common mechanism of gene inactivation during tumorigenesis. We examined the methylation status of two distinct regions of the MLH1 promoter (proximal and distal to the transcription start site) and the MLH1 gene expression by methylation-specific PCR and immunohistochemistry. A total of 72 colorectal tumors, both with (n = 51, 22 affected by hereditary nonpolyposis colorectal cancer, HNPCC, defined according to the international clinical criteria and 29 sporadic cases) and without microsatellite instability (MSI) (n = 21) were studied. Methylation was present in at least one of the two promoter regions in 86% of the sporadic MSI cases, in 33% of the cases lacking MSI, and in 23% of the HNPCC tumors. In the HNPCC cases with a known MLH1 mutation (n = 10) none of the two promoter regions was methylated. Hypermethylation in both MLH1 promoter regions was seen in 45% of the MSI sporadic cases vs. 5% of the MSI-negative cases and 0% of the HNPCC cases. The overall concordance between the two promoter regions regarding methylation status was good (P = 0.009), but no significant correlation between methylation and suppression of the MLH1 immunohistochemical expression was found. Our data confirm that mutation and hypermethylation are mutually exclusive mechanisms in inducing mismatch repair deficiency and support the hypothesis of methylation as a process evenly distributed along the different regions of the promoter.

2001 - Microsatellite instability and mismatch-repair protein expression in hereditary and sporadic colorectal carcinogenesis [Articolo su rivista]
Pedroni, Monica; Sala, E; Scarselli, A; Borghi, F; Menigatti, M; Benatti, Piero; Percesepe, Antonio; Rossi, Giorgio; Foroni, M; Losi, Lorena; Di Gregorio, C; DE POL, Anto; Nascimbeni, R; Di Betta, E; Salerni, B; PONZ DE LEON, Maurizio; Roncucci, Luca
abstract

Aberrant crypt foci (ACF) are microscopic clusters of altered colonic crypts considered premalignant lesions in the large bowel. Genomic instability at short tandem repeats in the DNA, referred to as microsatellite instability (MSI) is the hallmark of hereditary nonpolyposis colorectal carcinoma (HNPCC) caused by mutations in DNA mismatch-repair genes, mostly hMLH1 and LMSH2. In this study, we evaluated for MSI ACF (n = 16), adenomas (n = 18), carcinomas (n = 22), and lymph node metastases (n = 3) from 17 patients with colorectal cancer positive for MSI, Ten patients were members of HNPCC families; 7 patients had no family history of cancer. MSI was found in 7 of 7 (100%) ACF and 11 of 12 (91%) adenomas from patients with HNPCC, MSI was not related to histology and size of ACF. A progressive increase in instability as estimated by the number of shifted bands was observed along the ACF-adenoma-carcinoma sequence, In contrast, two of nine (228) ACF and none of six adenomas from patients with MSI sporadic carcinoma were unstable at microsatellite loci. hMLH1 or hMSH2 protein expression was altered only in MSI-positive premalignant lesions (ACF and/or adenomas), but not in all MSI-positive lesions in patients with HNPCC. These observations provide evidence of the premalignant nature of ACF in HNPCC and suggest that MSI is a very early event both in HNPCC and in sporadic colorectal carcinogenesis, although in the latter it seems infrequent.

2001 - Molecular screening for Hereditary Non Polyposis Colorectal Cancer (HNPCC): a prospective, population-based study [Articolo su rivista]
Percesepe, Antonio; F., Borghi; M., Menigatti; Losi, Lorena; M., Foroni; C., Di Gregorio; Rossi, Giuseppina; Pedroni, Monica; E., Sala; F., Vaccina; Roncucci, Luca; Benatti, Piero; A., Viel; M., Genuardi; G., Marra; P., Kristo; P., Peltomäki; PONZ DE LEON, Maurizio
abstract

PURPOSE: Germline mutations in mismatch repair genes predispose to hereditary nonpolyposis colorectal cancer (HNPCC). To address effective screening programs, the true incidence of the disease must be known. Previous clinical investigations reported estimates ranging between 0.5% and 13% of all the colorectal cancer (CRC) cases, whereas biomolecular studies in Finland found an incidence of 2% to 2.7% of mutation carriers for the disease. The aim of the present report is to establish the frequency of the disease in a high-incidence area for colon cancer.PATIENTS AND METHODS: Through the data of the local CRC registry, we prospectively collected all cases of CRC from January 1, 1996, through December 31, 1997 (N = 391). Three hundred thirty-six CRC cases (85.9% of the incident cases) were screened for microsatellite instability (MSI) with six to 12 mono- and dinucleotide markers. MSI cases were subjected to MSH2 and MLH1 germline mutation analysis and immunohistochemistry; the methylation of the promoter region was studied for MLH1.RESULTS: Twenty-eight cases (8.3% of the total) showed MSI. MSI cases differed significantly from microsatellite-stable (MSS) cases for their proximal location (P < .01), high mucinous component (P < .01), and poor differentiation (P = .002). Of MSI cases studied (n = 12), only one with a family history compatible with HNPCC had a germline mutation (in MSH2). Five other patients with a family history of HNPCC (two with MSI and three with MSS tumors) did not show germline mutations.CONCLUSION: We conclude that the incidence of molecularly confirmed HNPCC (one [0.3%] of 336) in a high-incidence area for CRC is lower than in previous biomolecular and clinical estimates.

2001 - Mutations of the minor mismatch repair gene MSH6 in typical and atypical HNPCC. [Articolo su rivista]
Lucci Cordisco, E.; Ravella, V.; Carrara, S.; Percesepe, Antonio; Pedroni, Monica; Bellacosa, A.; Caluseriu, O.; Fornasarig, M.; Neri, G.; PONZ DE LEON, Maurizio; Viel, A.; Genuardi, M.
abstract

Mutations of the mismatch repair (MMR) genes MLH1 and MSH2 are associated with hereditary nonpolyposis colorectal cancer (HNPCC), a highly penetrant autosomal dominant condition characterized by hypermutability of short tandemly repeated sequences in tumor DNA. Mutations of another MMR gene, MSH6, seem to be less common than MLH1 and MSH2 defects, and have been mostly observed in atypical HNPCC families, characterized by a weaker tumor family history, higher age at disease onset, and low degrees of microsatellite instability (MSI), predominantly involving mononucleotide runs. We have investigated the MSH6 gene sequence in the peripheral blood of 4 HNPCC and 20 atypical HNPCC probands. Two frameshift mutations within exon 4 were detected in 2 patients. One mutation was found in a proband from a typical HNPCC family, who had developed a colorectal cancer (CRC), a gastric cancer and a rectal adenoma. The CRC and the adenoma showed mild MSI limited to mononucleotide tracts, while the gastric carcinoma was microsatellite stable. The other mutation was detected in an atypical HNPCC proband, whose CRC showed widespread MSI involving both mono- and dinucleotide repeats. The phenotypic variability associated with MSH6 constitutional mutations represents a complicating factor for the optimization of strategies aimed at identifying candidates to MSH6 genetic testing.

2001 - The role of hPMS1 and hPMS2 in predisposing to colorectal cancer [Articolo su rivista]
Liu, T; Yan, H; Kuismanen, S; Percesepe, Antonio; Bisgaard, Ml; Pedroni, Monica; Benatti, Piero; Kinzler, Kw; Vogelstein, B; PONZ DE LEON, Maurizio; Peltomaki, P; Lindblom, A.
abstract

Hereditary nonpolyposis colorectal cancer (HNPCC) is attributable to a deficiency of mismatch repair. Inactivation of DNA mismatch repair underlies the genesis of microsatellite instability in colorectal cancer. Germline mutations in three DNA mismatch repair genes, hMSH2, hMLH1, and hMSH6, have been found to segregate in HNPCC and HNPCC-like families. The two DNA mismatch repair genes hPMS1 and hPMS2 have also been suggested to predispose to HNPCC. In this study, 84 HNPCC and HNPCC-like kindreds without known mutations in the other three known DNA mismatch repair genes were screened for germline mutations in the hPMS1 or hPMS2 gene. No clear-cut pathogenic mutations were identified. Conversion technology was used to detect a large hMSH2 deletion in two affected members of the kindred in which the hPMS1 mutation was originally reported, whereas the hRWS1 mutation was only present in one of these two individuals. Since the hPMS1 and hPMS2 genes were first reported, germline mutations in hPMS2 have been demonstrated primarily in patients with Turcot's syndrome. However, no mutation in any of the two genes has been found to segregate in HNPCC families. Until there is better evidence for an increased colorectal cancer risk associated with germline mutations in these genes, a conservative interpretation of the role of mutations in these genes is advised.

2001 - hMSH6 immunohistochemistry in families with clinical suspicion of Hereditary Non Polyposis Colorectal Cancer [Monografia/Trattato scientifico]
Scarselli, A.; Lucci Cordisco, E.; Benatti, Piero; Losi, Lorena; Menigatti, M.; Pedroni, Monica; Borghi, F.; Roncucci, Luca; Viel, A.; Genuardi, M.; PONZ DE LEON, Maurizio; Di Gregorio, C.
abstract

Not available

2000 - Aberrant crypt foci in colorectal carcinogenesis. Cell and crypt dynamics [Articolo su rivista]
Roncucci, Luca; Pedroni, Monica; F., Vaccina; Benatti, Piero; Marzona, Laura; DE POL, Anto
abstract

Aberrant crypt foci (ACF) have been identified on the colonic mucosal surface of rodents treated with colon carcinogens and of humans after methylene-blue staining and observation under a light microscope. Several lines of evidence strongly suggest that ACF with certain morphological, histological, cell kinetics, and genetic features are precursor lesions of colon cancer both in rodents and in humans. Thus, ACF represent the earliest step in colorectal carcinogenesis. This paper has the main purpose of reviewing the evidence supporting this view, with particular emphasis on cell and crypt dynamics in ACF. ACF have been used as intermediate biomarkers of cancer development in animal studies aimed at the identification of colon carcinogens and chemopreventive agents. Recently, evidence has also shown that ACF can be effectively employed in chemopreventive studies also in humans.

2000 - Genomic instability and target gene mutations in colon cancers with different degrees of allelic shifts [Articolo su rivista]
Percesepe, Antonio; Pedroni, Monica; Sala, E; Menigatti, M; Borghi, F; Losi, Lorena; Viel, A; Genuardi, M; Benatti, Piero; Roncucci, Luca; Peltomaki, P; PONZ DE LEON, Maurizio
abstract

Two grades (high and low) of microsatellite instability (MSI) are known, depending on the number of mutated markers and the amount of allelic shifts. Forty-two colorectal tumors, previously found to have high-degree MSI at dinucleotidic repeat loci, were revisited with BAT26, a mononucleotide marker, and the number of shifted bases were counted. Seven tumors, all with local stages at diagnosis, had less than or equal to 6-bp deletions and consistently displayed shorter shifts also with other intronic mononucleotide markers. Analysis of mononucleotide tracts in the coding regions of MSH3, MSH6, BAX, and TGF beta R11 in the groups with large (>6 bp) and short (less than or equal to 6 bp) allelic shifts showed specific patterns of involvement for the individual genes: TGF beta R11 displayed a uniformly high rate of mutations, while MSH3, MSH6, and BAX were less frequently altered in tumors with short shifts. Our findings suggest that microsatellite instability arises gradually, evenly involving loci with similar features of length and repetition. However, target genes have a specific timing of mutation in this process: TGF beta R11 is involved in the early phases, while BAX and MSH6 are frequently associated with big size shifts and tumors with more advanced stages.

2000 - Problems in the identification of hereditary nonpolyposis colorectal cancer in two families with late development of full-blown clinical spectrum [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Pedroni, Monica; A., Viel; M., Genuardi; Percesepe, Antonio; Roncucci, Luca
abstract

The recognition of Hereditary Nonpolyposis Colorectal Cancer (HNPCC) remains difficult despite the most recent advancements of molecular biology and technology. We describe two families with early onset of cancer but no suspicion of hereditary tumors; during follow-up, both families developed a tumor spectrum highly suggestive of HNPCC, thus emphasizing the importance of family history For a proper identification of hereditary tumors or cancer aggregation. Microsatellite instability was negative in tumors from both families and, as expected, no germline mutations of the major DNA mismatch repair genes (MSH2 and MLH1) could be detected. Suspicion of the disease at the time of proband´s lesion might have led to prevention, or early diagnosis, of at least three malignant turners. We conclude that a possible genetic origin should always be suspected in individuals with early-onset neoplasms of the large bowel and probably of other organs such as the endometrium, small bowel, and urothelium, even when the initial pedigree does not show marked aggregation of cancers or vertical transmission.

2000 - Staging and survival of colorectal cancer: are we making progress? The 14-year experience of a Specialized cancer Registry [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Di Gregorio, C; Fante, R; Rossi, Giorgio; Losi, Lorena; Pedroni, Monica; Percesepe, Antonio; Roncucci, Luca
abstract

Background and Aims. It is still unclear whether recent advancements in colorectal cancer research have led to an improvement in management and prognosis of the disease. Through the data of a specialized colorectal cancer Registry we aimed at analysing pathological staging and 5-year survival of ail patients with malignancies of large bowel diagnosed between 1984 and 1997. Main objective was to ascertain whether or not we are making progress in the control of this common neoplasm. Patients and Methods. During the 14-year period 1984-97, a total of 2,240 colorectal cancer patients were registered, for a crude incidence rate of 64.5 and 55.2/100,000/year in males and females, respectively Tumours were staged with Tumour Node, Metastasis system, corresponding to Dukes' classification, into four main groups. Survival was assessed with Life Table analysis, and statistical significance - between various subgroups - evaluated with Log-Rank Test. Results. Crude incidence rates of colorectal neoplasms showed minor fluctuations during initial period of registration, increasing sharply after 1990 mainly due to localized (stage I and II) lesions and, to a lesser degree, to stage ill tumours. Number of advanced (stage IV and unstaged) malignancies remained virtually stable. When results were expressed as percent of total cases, the fraction of localized lesions increased from 39% in the biennium 1984-5 to 51.6% in 1986-97, and the proportion of advanced tumours fail from 39% to 21.6% (p for trend <0.001). As expected, 5-year survival was significantly (p<0.002) more favourable for individuals diagnosed in 1990-91 than for patients registered in 1984-89. Conclusions. In Northern Italy, incidence rates of colorectal carcinoma are rising. This trend is associated with a sharp increase of newly detected localized lesions and with a significant improvement of overall 5-year survival The result may be attributed to several concomitant factors, such as: A) wider use of colonoscopy, B) increased education of patients, C) more attention given to symptoms.

1999 - Clinical and molecular diagnosis of hereditary non-polyposis colorectal cancer: problems and pitfalls in an extended pedigree [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Percesepe, Antonio; Rossi, Giorgio; A., Viel; M., Santarosa; Pedroni, Monica; Roncucci, Luca
abstract

Hereditary non-polyposis colorectal cancer (or Lynch syndrome) is an autosomal dominant disease in which early onset colorectal carcinomas aggregate in families together with tumours of other organs. The genetic basis of the syndrome has been clarified with the identification of mutations in several DNA mismatch repair genes (MSH2, MLH1, PMS1, PMS2 and MSH6). We describe the clinical features and molecular characterization of a large hereditary nonpolyposis colorectal cancer family which has been followed for almost 10 years. The kindred showed a striking aggregation of colorectal tumours in 3 successive generations; most of these neoplasms developed before the age of 50 years and were localized in the proximal colon. Molecular tests (carried out in ten individuals) showed specific alterations at the MLH1 gene, consisting in the insertion of a T nucleotide between bases 2,269 and 2,270; the mutation caused frameshift of the open reading frame and synthesis of a polypeptide longer than normal. The only tumour that could be analysed was positive for microsatellite instability. Physicians should become more confident with hereditary tumours and their implications, which are not limited to a single individual but concern all family members at risk of cancer This family approach is different, and requires more expertise than the traditional individual approach. Common problems encountered in Hereditary Non-polyposis Colorectal Cancer families include: A) poor collaboration of subjects at risk (a situation which may cause some conflict between the doctor's duty to inform patients about their risk of disease and the rights of patients to choose and decide about their health); B) definition of the most appropriate surveillance programme for a given family (how many investigations to propose to the patients, and how often); C) possible interaction between genes and environmental factors (for instance, a gene carrier - in this family - developed an endometrial carcinoma after standard tamoxifen adjuvant therapy for breast cancer).

1999 - Epidemiologic and genetic factor in colorectal cancer: development of cancer in dizygotic twins in a family with Lynch syndrome [Articolo su rivista]
PONZ DE LEON, Maurizio; Pedroni, Monica; Benatti, Piero; Percesepe, Antonio; Rossi, Giorgio; M., Genuardi; Roncucci, Luca
abstract

Human tumours usually develop due to a close inter action between environmental and genetic factors. This concept applies also to well defined genetic diseases such as Hereditary Nonpolyposis Colorectal Cancer (HNPCC or Lynch syndrome), which is featured by early onset tumours of the large bowel(and other target organs), striking aggregation of neoplasms in families, and vertical transmission consistent with an autosomal dominant pattern of inheritance. As a further example of gene/environment interaction, we report on a Hereditary Nonpolyposis Colorectal Cancer family in which two dizygotic twins were affected by cancer of the large bowel. One of the twins was slightly overweight and showed many common risk factors for colorectal carcinoma he developed a Dukes' C lesion at the age of 52 The other twin was not overweight and was much less exposed to exogenous risk factors; a Dukes' B carcinoma was diagnosed nt age 60 during a control endoscopy. This anedoctal report suggests that diet and lifestyle are of relevance also in patients with genetically determined tumours of the large bowel. It follows that the control of these environmental factors might be associated with a delay of tumour occurrence and possibly with a less aggressive tumour behaviour.

1999 - Epidemiology of cancer of the large bowel - The 12-year experience of a specialized registry in Northern Italy [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Percesepe, Antonio; C., Di Gregorio; R., Fante; Losi, Lorena; Rossi, Giorgio; Pedroni, Monica; Roncucci, Luca
abstract

Background. In 1984 a specialized colorectal cancer registry was instituted in Modena; aims of the Registry were: the evaluation of incidence and mortality, the study of morphological aspects, staging, staging and familiarity of the registered patients. Aims. Purpose of the research was to provide an updated description of the main findings (in particular incidence, staging, morphology and survival) observed in the 12-year registration period. Patients and methods. Between January 1984 and December 1995, 1,899 malignancies of the large bowel in 1,831 patients were registered. Tumours were classified according to the International Classification of the Diseases for Oncology (ICD-O) and staged with the TNM system Cancer specific srm il nl was assessed with life table analysis and Log-Rank tests. Results. Crude incidence rare showed minor fluctuations between 1984 and 1989, but tended to rise in the following pears. Tumours were mostly located distal to the splenic flexure (73.3% of the total), with a slight tendency over time to a gradual shift to the right colon. Staging became progressively more favourable throughout the registration; in 1984 both stages I, II and stage IV + unstaged lesions represented 40% of the total, but in 1995 the former rose to 50% whereas the latter fell to 21.6% (p<0.001). This more to earlier stages resulted in an improved survival of patients registered in 1990-91 versus 1983-85 (Log-Rank 14.3 p<0.002). Factors associated with a poor survival were the advanced age of patients at diagnosis (>74) and clinical stage. Conclusions. Main aspects of the investigation were the increasing crude incidence rates of colorectal turnovers and the gradual increase of neoplasms diagnosed irt a more favourable staging. It is like likely that the improvement of staging and survival observed in the 12 fears of registration can be attributed to the improved attitude to health care of the population, and possibly to the improvement of surgical techniques.

1999 - Evaluation of the replication error phenotype in relation to molecular and clinicopathological features in hereditary and early onset colorectal cancer. [Articolo su rivista]
Capozzi, E; Della Puppa, L; Fornasarig, M; Pedroni, Monica; Boiocchi, M; Viel, A.
abstract

Mutations affecting human mismatch repair (MMR) genes (MLH1, MSH2, PMS1, PMS2, and MSH6) cause tumour predisposition in hereditary nonpolyposis colorectal cancer (HNPCC) syndrome, and an association has been demonstrated with the replication error (RER) phenotype in most colorectal and some extracolonic neoplasms. A pathogenetic model for RER+ tumours through inactivation of suppressor genes has been hypothesised, and TGF beta RII, BAX and IGFIIR genes have recently been proposed as targets of such inactivating mutations. In this study, a series of 47 tumours developed in patients with known MLH1/MSH2 status and a family history of HNPCC and/or early onset colorectal cancer were characterised for the RER phenotype through microsatellite analysis. The RER phenotype, displayed by 17 tumours, was then correlated with the presence of insertions/deletions at the TGF beta RII, IGFIIR and BAX gene stretches, confirming that the TGF beta RII inactivation may be particularly critical for the RER-associated tumorigenesis. RER+ colorectal cancers (CRCs) developed more frequently in patients from HNPCC families (72.7%) than in those from families not fulfilling the Amsterdam criteria (33.3% in suspected HNPCC and 20.8% in early onset CRC patients). A consistent fraction of either Amsterdam and non-Amsterdam patients developed RER- CRCs, pointing to the involvement of other genes not related to the MMR system. The RER phenotype was associated with younger age at diagnosis in familial cases, and there was a trend for an association with proximal CRC localisation and early Dukes' stages. The RER status was also correlated with the presence and type of MLH1 and MSH2 alteration.

1999 - Hereditary colorectal cancer in the general population: from cancer registration to molecular diagnosis [Articolo su rivista]
PONZ DE LEON, Maurizio; Pedroni, Monica; Benatti, Piero; Percesepe, Antonio; Di Gregorio, C; Foroni, M; Rossi, Giorgio; Genuardi, M; Neri, G; Leonardi, F; Viel, A; Capozzi, E; Boiocchi, M; Roncucci, Luca
abstract

Background-Hereditary non-polyposis colorectal cancer (HNPCC) is one of the most common inherited disorders predisposing to cancer. The genes responsible for the disease have recently been cloned and characterised; their mutations induce a generalised genomic instability which is particularly evident at microsatellite loci (replication error (RER)+ phenotype). Aims-To investigate how to select individuals and families in the general population who should be screened for constitutional mutations predisposing to colorectal cancer. Patients/Methods-Between 1984 and 1995, 1899 colorectal malignancies in 1831 patients were registered, and in 1721 of these (9-1%), family trees could be obtained. Patients and families were classified into five categories according to a more or less likely genetic basis: HNPCC; suspected HNPCC;; juvenile cases; aspecific cancer aggregation; sporadic cases. In 18 families with HNPCC as well as in 18 with suspected HNPCC, microsatellite instability in tumour tissues and constitutional mutations of two DNA mismatch repair genes (MSH2 and MLH1) could be evaluated. RER status was studied with five markers (BAT40, D2S123, D18S57, D17S787, and BAT26) in paraffin embedded tissues. Germline mutations of MSH2 or MLH1 genes were assessed on DNA and RNA extracted from lymphomonocytic cells, using reverse transcription polymerase chain reaction, single strand conformation polymorphism analysis, and direct DNA sequencing. Results-HNPCC represented 2.6% and suspected HNPCC 4.6% of all registered colorectal neoplasms. Eleven out of 18 HNPCC families (61%) showed microsatellite instability as opposed to four (of 18) suspected HNPCC (22%; p<0.02). Three germline mutations (two in MSH2 and one in MLH1 gene) were found in three different large HNPCC families, whereas no mutations were detected in suspected HNPCC. Conclusions-In this study of cancer genetic epidemiology, data from a tumour registry were analysed and this ultimately led to the identification and selection of families that should be tested for mutator gene mutations. With the use of a population based approach, the incidence of mutations was appreciably lower than previously reported and limited to families with full blown HNPCC. It is possible that in most families with a clinical spectrum Of HNPCC (or suspected HNPCC) other DNA mismatch repair genes are involved in the pathogenesis of the disease.

1999 - Microsatellite instability in multiple colorectal tumors [Articolo su rivista]
Pedroni, Monica; Mg, Tamassia; Percesepe, Antonio; Roncucci, Luca; Benatti, Piero; G., Lanza; R., Gafa; C., Di Gregorio; R., Fante; Losi, Lorena; L., Gallinari; PONZ DE LEON, Maurizio; F., Scorcioni; F., Vaccina; Rossi, Giuseppina; A. M., Cesinaro
abstract

Tumor multiplicity is a hallmark of hereditary cancers: in the colon-rectum multiple tumors represent 5-10% of all colorectal cancer cases. A portion of these cases belongs to hereditary non-polyposis colorectal cancer (HNPCC), a genetic cancer syndrome due to mismatch repair (MMR) gene mutations, phenotypically expressed as microsatellite instability (MSI); the majority of multiple tumors, however, is apparently without any family history. We analyzed 78 (38 synchronous and 40 metachronous) neoplasms from 37 patients with multiple tumors of the large bowel, both HNPCC and sporadic, with the aim of identifying a common genetic basis in multiple tumors. DNA was extracted from normal and cancerous formalin-fixed tissue and was analyzed for MSI using 6 markers. Tumors showing MSI in at least 2 of 6 microsatellite loci were defined as MSI(+). The overall number of MSI(+) tumors was 22 (28.2% of the total). A significant difference in the rate of MSI(+) between HNPCC and sporadic tumors was observed (85% vs. 17%). In the same patients, the MSI phenotype of synchronous tumors (both HNPCC and sporadic) tended to be more concordant than that of the metachronous ones. The higher frequency of MSI in HNPCC than in sporadic tumors, even when multiple, suggests that the involvement of MMR genes in the pathogenesis of the sporadic cases may be uncommon, thus confirming that screening for MSI in multiple colorectal tumors could be a useful tool in the identification of HNPCC in the general population.

1998 - Aberrant crypt foci in patients with colorectal cancer [Articolo su rivista]
Roncucci, Luca; S., Modica; Pedroni, Monica; Mg, Tamassia; M., Ghidoni; Losi, Lorena; R., Fante; C., Di Gregorio; Manenti, Antonio; L., Gafa; PONZ DE LEON, Maurizio
abstract

Aberrant crypt foci (ACF) are clusters of abnormally large colonic crypts identified on the mucosal surface of the human colon. They are thought to be preneoplastic lesions. The aim of the present study was to compare density (number of ACF per square cm of mucosal surface), crypt multiplicity (number of crypts per ACF) and histology of ACF in colonic resections of colorectal cancer patients resident in two Italian provinces with a twofold difference in colorectal cancer incidence rates. Thirty-two and 26 colonic resections were collected after operation in Ragusa (Southern Italy) and Modena (Northern Italy), respectively, and fixed in 10% formalin. Mucosal layers were observed under a light microscope at 25x after staining with methylene blue. Density of ACF was significantly higher in Modena (median 0.101 ACF cm(-2)) than in Ragusa (0.049, P= 0.001), whereas there was no difference in crypt multiplicity. ACF were classified into three groups according to histological features. ACF with mild alterations (hypertrophic ACF, 73%), ACF with hyperplasia (hyperplastic ACF, 17%) and ACF with dysplasia (microadenomas, 10%). The proportions of ACF in the three groups were similar in the two provinces, Density of ACF was higher and crypt multiplicity lower proceeding from proximal to distal large bower. Microadenomas were observed only in the colon, whereas hyperplastic ACF were more frequent in the rectum, In conclusion, density of ACF correlates with colorectal cancer rates in two Italian provinces, and shows a positive gradient from proximal to distal large bowel. Histology of ACF suggests that they may be precursors of both hyperplastic and adenomatous polyps. These data provide further evidence of the role of ACF in human colorectal carcinogenesis.

1998 - Characterization of MLH1 and MSH2 alternative splicing and its relevance to molecular testing of colorectal cancer susceptibility. [Articolo su rivista]
Genuardi, M; Viel, A; Bonora, D; Capozzi, E; Bellacosa, A; Leonardi, F; Valle, R; Ventura, A; Pedroni, Monica; Boiocchi, M; Neri, G.
abstract

The phenomenon of alternative splicing in the DNA mismatch repair genes MLH1 and MSH2 was extensively investigated by coupled reverse transcription-polymerase chain reaction in different human tissues, including 42 mononuclear blood cell samples--31 obtained from familial colon cancer patients or their at-risk relatives and 11 from healthy blood donors--7 normal colonic mucosae, 4 established human cancer cell lines, 8 colorectal tumors, and one sample each of ileum, liver, muscle, thymus, breast, and EBV-transformed lymphoblasts. Several isoforms were observed for each gene. Products of MLH1 alternative splicing included mRNAs lacking alternative exons 6/9, 9, 9/10, 9/10/11, 10/11, 12, 16, and 17. For MSH2, products lacking exons 5, 13, 2 through 7, and 2 through 8 were identified. The levels of expression were found to vary among different samples. All isoforms were found in a relevant fraction (43-100%) of the mononuclear blood cell samples, as well as in other tissues. The splicing variants were also detected in normal colonic mucosa, with the exceptions of the MLH1 -6/9 and -10/11 and the MSH2 -13 isoforms. Germline mutations of MLH1 and MSH2 confer constitutional predisposition to the development of colorectal cancer and other neoplasms. A substantial proportion of the mutations identified so far involve alterations of the normal splicing process. Knowledge of the existence of multiple alternative splicing events, not caused by genomic DNA changes, is important for the evaluation of the results of molecular diagnostic tests based on RNA analysis.

1998 - Hereditary nonpolyposis colorectal cancer: an approach to the selection of candidates to genetic testing based on clinical and molecular characteristics [Articolo su rivista]
A., Viel; M., Genuardi; E., LUCCI CORDISCO; E., Capozzi; V., Rovella; M., Fornasarig; PONZ DE LEON, Maurizio; M., Anti; Pedroni, Monica; A., Bellacosa; Percesepe, Antonio; M., Covino
abstract

Objective: Identification of clinical and molecular characteristics associated with constitutional MLH1 and MSH2 mutations and definition of a stepwise strategy for the selection of colorectal cancer (CRC) patients amenable to MLH1 and MSH2 genetic testing. Methods: 90 unrelated CRC patients were initially selected on the basis of either familial or early onset occurrence of CRC. They were screened for the presence of constitutional MLH1 and MSH2 mutations and for microsatellite instability (MSI). Results: 16 pathogenetic mutations (9 MLH1 and 7 MSH2) were identified in 41% of Amsterdam hereditary nonpolyposis colorectal cancer (HNPCC) families, 5% of suspected HNPCC families, and 14% of sporadic early-onset CRC patients. The presence of the mutations correlated with MSI, with early age of onset and proximal location of the tumor, and with the presence of some extracolonic tumors of the HNPCC spectrum and/or multiple tumors in the family. Conclusions: Evaluation of clinical and molecular characteristics is useful for the identification of candidates to MLH1 and MSH2 mutational analysis and allows the application of a rational approach to genetic testing.

1998 - Lack of PMS2 gene-truncating mutations in patients with hereditary colorectal cancer [Articolo su rivista]
Viel, A; Fornasarig, M; Novella, E; Genuardi, M; Capozzi, E; Pedroni, Monica; Santarosa, M; PONZ DE LEON, Maurizio; Della Puppa, L; Anti, M; Boiocchi, M.
abstract

Hereditary non-polyposis colorectal cancer (HNPCC) is a genetically heterogeneous disease for which PMS2 gene, a member of the human PMS gene family, is believed to have a marginal role. To better define the contribution of PMS2 to hereditary colorectal cancer, we investigated this gene in 22 unrelated Italian patients that, despite a positive family history and/or early onset and development of tumors with microsatellite instability (MSI), did not carry constitutional mutations of MLH1 and MSH2 genes. No mutations with clear-cut pathogenetic significance were detected in the coding regions of PMS2 gene, but only 8 polymorphisms (7 common and 1 rare, 3 silent and 5 missense) and 3 unique molecular variants (2 missense substitutions and one 3-nucleotide deletion) were seen. Lack of PMS2 truncating mutations in our study does not disagree with its supposed marginal involvement in hereditary colorectal cancer, but at the same time points out the need to investigate the phenotypic molecular and clinical characteristics more specifically associated with PMS2 mutations.

1998 - Scanning electron microscopy of aberrant crypt foci in human colorectal mucosa [Articolo su rivista]
F., Vaccina; F., Scorcioni; Pedroni, Monica; Mg, Tamassia; PONZ DE LEON, Maurizio; DE POL, Anto; Marzona, Laura; Roncucci, Luca
abstract

Background: Aberrant crypt foci (ACF) are clusters of morphologically altered crypts which can be observed by light or stereomicroscopy on the mucosal surface of the colon after staining with methylene-blue. They probably represent one of the earliest events in human colorectal carcinogenesis. The main purpose of the present study was to observe the surface features of aberrant and normal colonic crypts in humans using scanning electron microscopy (SEM) in order to find and measure differences between aberrant and normal. Materials and Methods: Fifteen mucosal specimens containing ACF and 8 with normal mucosa taken from patients operated on for colon cancer were observed under a scanning electron microscope. Results: By SEM ACF were easily observed on the mucosal surface because they showed a well defined border and were elevated on the mucosal surface. Under higher magnification luminal openings of aberrant crypts had a larger overall average diameter than normal (37.6 mu m +/- 13.5, mean a SD, vs 15.9 mu m +/- 4.9, P=0.001), though when crypt multiplicity of ACF (number of crypts per ACF) was higher, the diameter of luminal openings tended to be smaller and similar to those of normal crypts, with weak negative correlation between crypt multiplicity of ACF and mean diameter of aberrant luminal openings (r=-0.27). Finally, the mucosal surface among aberrant crypts was flattened because of a loss of microvilli. In conclusion, scanning electron microscopy allows a better definition of the topological features of aberrant crypt foci than light or stereomicroscopy.

1998 - Small bowel carcinoma in hereditary nonpolyposis colorectal cancer [Articolo su rivista]
Benatti, Piero; Roncucci, Luca; Percesepe, Antonio; A., Viel; Pedroni, Monica; Mg, Tamassia; F., Vaccina; R., Fante; S., De Pietri; PONZ DE LEON, Maurizio
abstract

A 53-yr-old man, a member of a hereditary nonpolyposis colorectal cancer (HNPCC) family, with previous colonoscopic polypectomies, presented for persisting vomiting and marked signs of dehydration. Previous radiological and endoscopic examinations of the upper digestive tract were negative, with the exception of the presence of a duodenal adenomatous polyp. Enteroclysis led to a diagnosis of obstruction at the Treitz angle due to a moderately differentiated adenocarcinoma, Microsatellite instability was demonstrated in the DNA extracted from the tumor. The patient was the carrier of a mutation in the intron 13 of the hMLH1 gene, one of the four mismatch repair genes known to be responsible for HNPCC.

1997 - Colorectal carcinoma in different age groups: A population based investigation [Articolo su rivista]
R., Fante; Benatti, Piero; S., Depietri; Pedroni, Monica; Mg, Tamassia; Percesepe, Antonio; Losi, Lorena; Roncucci, Luca; PONZ DE LEON, Maurizio; DI GREGORIO, Carmela; Rossi, Giuseppina
abstract

Although colorectal cancer is a disease of the older population, these tumors are not infrequent before the age of 55. Through the data of a population-based registry, we proposed giving a description of the clinical features of three groups of patients in whom the disease occurred at a relatively early age of onset (group I: < 40 yr; group II: 41-50 yr; group III: 51-55 yr). There were only 14 patients under the age of 40 yr (1.1% of total registered patients, n = 1298 in the period 1984-1992). Group II and III represented 5.9% and 6.0%, respectively (n = 76 and 78), with minor fluctuations throughout the 9-yr period of registration. Inherited colorectal tumors [hereditary nonpolyposis colorectal cancer (HNPCC), adenomatosis coli, and suspected HNPCC] accounted for 38.4% of group I patients (5 of 14), 17.1% of group II, 10.2% of group III, and only 3.5% of individuals older than 55 (p, for trend, < 0.001). Thus, hereditary colorectal tumors were detected significantly more often in younger individuals. The majority of colorectal malignancies were localized in the left colon or rectum in all three groups,,vith a tendency (not significant) to a preferential localization in the right colon for tumors developed in group I (37% vs 18% and 14% in groups II and III, respectively). Pathological stage and main histological types did not differ among the three groups. Finally, life-table analysis did not show significant differences in 5-yr survival among the three groups; however, when considered together, early onset cases showed a more favorable prognosis than older individuals (log-rank 11.6; p < 0.001). In conclusion, colorectal cancer is diagnosed very rarely before the age of 40 yr, whereas about 12% of all cases belong to the age group 41 to 55 yr of age. Hereditary tumors were found more frequently in younger patients, with a well-defined inverse relationship between age of onset and frequency of genetically determined tumors. Finally, the clinical outcome was more favorable in the whole series of early onset cases than in older registered patients.

1997 - Histology of aberrant crypt foci in the human colon [Articolo su rivista]
Di Gregorio, C; Losi, Lorena; Fante, R; Modica, S; Ghidoni, M; Pedroni, Monica; Tamassia, Mg; Gafà, R; PONZ DE LEON, Maurizio; Roncucci, Luca
abstract

Aberrant crypt foci (ACF) have been identified in the methylene-blue stained mucosa of the human colon, Some lines of evidence suggest that ACF may be precursors of colon cancer, The objective of the present study was to establish morphological criteria able to define and classify ACF in histological sections, Twenty four colectomy specimens were collected after operation for colorectal cancer and fixed in 10% formalin, Strips of grossly normal mucosa were stained in a 0.2% solution of methylene blue in saline for 5-10min. The strips were measured, put on a glass slide and observed under a light microscope at x25, One hundred and fourteen ACF identified by topology were sectioned parallel to the muscularis mucosae. Eighty-four ACF were evident at histological examination and could be classed into three main groups: group A (61 ACF 72.6%) including foci whose epithelial cells had regular nuclei, with only mild or focal crowding but no stratification, no mucin depletion and no dysplasia; group B (16 ACF 19.1%), in which features of hyperplasia were evident; and group C (seven ACF 8.3%) including foci with enlarged, crowded and stratified nuclei, mucin depletion, frequent mitoses, and evident dysplasia, diffuse or focal (mild in five cases, moderate in two) representing microadenomas, Finally, hyperplastic foci were significantly larger than foci of group A and C, Group B ACF were also more frequent in the rectum than in the colon. In conclusion, selected histological features allow the definition of groups of ACF, which may represent sequential steps in the development of human colorectal tumours.

1997 - K-ras and p53 mutations in hereditary non-polyposis colorectal cancers [Articolo su rivista]
Losi, Lorena; PONZ DE LEON, Maurizio; Jiricny, J; Di Gregorio, C; Benatti, Piero; Percesepe, Antonio; Fante, R; Roncucci, Luca; Pedroni, Monica; Benhattar, J.
abstract

Genetic instability related to defective DNA mismatch repair genes may be involved in the pathogenesis of carcinoma in Hereditary Non-Polyposis Colorectal Cancer (HNPCC). To test that the targets of genetic instability could include critical transforming genes involved in colon tumor progression, we examined 23 colorectal carcinomas in patients with HNPCC in order to detect somatic mutations in K-ros and p53 genes. Using single strand conformation polymorphism followed by direct DNA sequencing, we detected 4 mutations in K-ros gene (17%) and 3 in p53 gene (13%) which change the aminoacid sequence of the protein p53. This is significantly lower than in sporadic cancer. Our data suggest that colon cancer in HNPCC might partly involve a distinct pathogenetic mechanism that involves other genes than those altered in sporadic tumors.

1996 - Clinical features, frequency and prognosis of Dukes' a colorectal carcinoma: A population-based investigation [Articolo su rivista]
C., Digregorio; R., Fante; Roncucci, Luca; Mg, Tamassia; Losi, Lorena; Benatti, Piero; Pedroni, Monica; Percesepe, Antonio; S., Depietri; PONZ DE LEON, Maurizio
abstract

The main aim of this study was, through the data of a population-based Registry, to establish the incidence of Dukes´ A lesions by year of registration and the main clinical features, and to assess cancer-specific survival. One hundred and eighteen Dukes´ A colorectal tumours were diagnosed (in 117 patients) out of 1337 registered between 1984 and 1992 in the Health Care District of Modena, Northern Italy; 94 patients were treated with surgery and 23 with endoscopic polypectomy. The frequency of Dukes´ A tumours ranged between 4.8% and 18% by year of registration. Dukes´ A carcinomas were significantly more frequent in the distal colon. Only 5 patients (4%) died of their cancer, and in all patients the tumour was localised in the rectum. Carcinomas associated with a poor prognosis did not show any of the biological variables usually associated with an unfavourable outcome, but, our data suggest the possibility of incomplete removal of tumours at surgery. Copyright

1996 - Frequency and clinical features of multiple tumors of the large bowel in the general population and in patients with hereditary colorectal carcinoma [Articolo su rivista]
Fante, R; Roncucci, Luca; Di Gregorio, C; Tamassia, Mg; Losi, Lorena; Benatti, Piero; Pedroni, Monica; Percesepe, Antonio; De Pietri, A; PONZ DE LEON, Maurizio
abstract

BACKGROUND. Reports on the frequency of multiple carcinomas of the colon and rectum have varied from 3-4% to more than 10% of all tumors of the large bowel. METHODS. We reviewed the files of a specialized colorectal cancer registry with chronous or metachronous) in the general population; b) to compare these values with those observed in patients with hereditary nonpolyposis colorectal carcinoma the following objectives: a) to determine the frequency of multiple tumors (synchronous or metachronous) in the general population; b) to compare these values with those observed in patients with hereditary nonpolyposis colorectal carcinoma (HNPCC); and c) to evaluate the clinical outcome of patients with multiple tumors and the role of other clinical parameters in the development of these neoplasms. RESULTS. From 1984 to 1992, 53 patients with multiple tumors (of 1298 registered patients, 4%) had large bowel carcinoma; 33 (2.5%) were synchronous and 20 (1.5%) metachronous. The total number of multiple colorectal carcinomas was 95, which was 7% of all registered colorectal carcinomas (1337 carcinomas in 1298 patients). Multiple tumors occurred significantly more often in patients with HNPCC than in those with sporadic carcinomas (P < 0.001); this increased prevalence was more marked for metachronous lesions, which occurred almost 4 times more often in patients with HNPCC (5.8% vs. 1.3%; P < 0.001). The average interval of time between the first and the second malignancy was 8.7 years; there was no significant difference between hereditary and sporadic tumors. Patients with synchronous tumors did not show appreciable differences in survival when compared with individuals who had single neoplasms. In contrast, a poor clinical outcome was observed in patients with metachronous tumors after the development of the second carcinoma. Finally, polypoid adenomas of the large bowel were found significantly more often in patients with multiple primary tumors than in those with a single tumor. CONCLUSIONS. These results emphasize the importance of preoperative pancolonoscopy for the identification of possible synchronous tumors (both benign and malignant) and long-lasting endoscopic follow-up for the detection of recurrent or metachronous lesions. The conclusions are even more pertinent for patients with HNPCC, whose risk of metachronous tumors is significantly higher than that of patients with sporadic carcinoma.

1995 - Argyrophilic nucleolar organizer regions and bromodeoxyuridine and h3-thymidine labelling indices in colorectal cancer [Articolo su rivista]
Losi, Lorena; C., Di Gregorio; R., Fante; Migaldi, Mario; Roncucci, Luca; Pedroni, Monica; PONZ DE LEON, Maurizio; G. P., Trentini
abstract

The count of argyrophilic nucleolar organizer regions (AgNORs) has been proposed as a useful method for evaluating cell replication in human tumours. The current study was undertaken to compare AgNOR values in colorectal cancers with two better established methods for investigating cell proliferation such as bromodeoxyuridine (BrdUrd) and (3)[H]-thymidine ((3)[H]dT) labelling indices (LIs). Because some concern still exists regarding accuracy and reproducibility of AgNOR quantifying methods, we carried out a control study by independently repeating the same measurements (number, area and area per silver-stained NOR particle) in two centres with different operators and computer-assisted image analysers on 40 colorectal carcinomas. AgNOR values recorded in the two centres were strictly correlated (r = 0.75; P &lt; 0.001 for number; r = 0.62, P &lt; 0.01 for area; r = 0.63, P &lt; 0.001 for area per silver-stained NOR particle) and the range of values were almost identical, Then, AgNOR values were compared with BrdUrd and (3)[H]dT LIs, respectively obtained by in vivo incorporation and in vitro incubation in the same series of colorectal carcinomas. No correlation was found between AgNOR values and BrdUrd or (3)[H]dT LIs. BrdUrd and (3)[H]dT LIs were instead reciprocally significantly correlated, No evident correlation was seen between LIs or AgNOR values and clinico-pathological parameters of the tumour. In conclusion, in colorectal neoplasms, AgNOR values did not appear to relate with more direct parameters of cell proliferation. It follows that AgNOR reliability as a biomarker of cell proliferation remains questionable.

1995 - Biologic characterization of Hereditary Non-Polyposis Colorectal Cancer. Nuclear ploidy, AgNOR count, microvessel distribution, oncogene expression, and grade-related parameters. [Articolo su rivista]
Losi, Lorena; R., Fante; C., DI GREGORIO; M. L., Aisoni; G., Lanza; I., Maestri; Roncucci, Luca; Pedroni, Monica; PONZ DE LEON, Maurizio
abstract

The identification of hereditary non-polyposis colorectal cancer (HNPCC) is important not only for the patient, but also for family members who are at increased risk of developing cancer. To determine if measuring various pathobiologic features of the colon carcinomas is useful in separating sporadic from HNPCC tumors, the authors studied tumor tissues from 46 patients with HNPCC and compared them to 70 with sporadic colorectal carcinoma. Parameters investigated included DNA ploidy (flow cytometry), AgNOR count (by silver staining), microvessel density (immunohistochemistry), p53 and K-vas expression, and grade-related parameters. Diploid tumors were more frequent in patients with HNPCC (65% vs 40%, P <.02), thus confirming previous observations concerning such an association. Higher AgNOR counts and greater AgNOR areas were observed in sporadic tumors than in HNPCC (5.2 +/- 1.5 vs 4.5 +/- 1.8, P <.01). Hereditary tumors tended to be less vascularized, whereas oncogene expression and grade-related parameters did not show appreciable differences between the two types of tumors. In conclusion, some of the investigated parameters may contribute to defining the biologic profile of HNPCC. In addition, these findings support the clinical impression of a more favorable outcome that is frequently seen in HNPCC patients.

1995 - RISK OF CANCER REVEALED BY FOLLOW-UP OF FAMILIES WITH HEREDITARY NONPOLYPOSIS COLORECTAL-CANCER - REPLY [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Pedroni, Monica; R., Sassatelli; Roncucci, Luca
abstract

N/A

1993 - Cell kinetic evaluation of human colonic aberrant crypts [Articolo su rivista]
Roncucci, Luca; Pedroni, Monica; Fante, R; DI GREGORIO, C; PONZ DE LEON, Maurizio
abstract

Foci of aberrant crypts (ACF) have been observed on the unsectioned, methylene blue-stained mucosal surface of the human colon. Experimental evidence and the histological features of the lesions suggest that they might be early events in colon cancer development. The main objective of the present study was to evaluate cell kinetic properties of ACF in the human colon. Five samples of colon mucosa were collected immediately after operation following the administration of 500 mg of 5'-bromo-2'-deoxyuridine prior to surgery. ACF were then identified on the fixed, unsectioned, methylene blue-stained mucosal surface under a light microscope. Some specimens containing ACF were serially sectioned perpendicular to the luminal surface of the intestine, along with specimens of normal-appearing mucosa. Several sections were prepared for the immunohistochemical identification of 5'-bromo-2'-deoxyuridine-incorporating cells (in the S phase of the cell cycle). The results of this study demonstrated that aberrant crypts have more cells per crypt than normal glands. Total labeling index and labeling index values in each of the five longitudinal compartments in which each crypt was divided showed an increased total proliferative activity in all ACF examined, although limited to the lower crypt compartments in almost all aberrant crypts evaluated. These findings are in keeping with previous cell kinetic studies and observations in experimental animals and provide evidence of the involvement of human aberrant crypts in the stepwise process leading from normal mucosa to colon cancer.

1993 - Risk of cancer revealed by follow-up of families with hereditary non-polyposis colorectal cancer: a population-based study [Articolo su rivista]
PONZ DE LEON, Maurizio; Benatti, Piero; Pedroni, Monica; R., Sassatelli; Roncucci, Luca
abstract

Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal dominant disorder characterized by susceptibility to large-bowel cancer, frequently with early onset and localized in the right colon. Several tumours of other sites may also occur with increased frequency in these families. During the period 1984-1989, 28 families with HNPCC were identified in our Health Care District through a population-based colorectal cancer registry. Moreover, 61 additional families were selected and classified as having ´´suspected´´ HNPCC. The objective of the present study is to report the occurrence of new cases of cancer in the 5- to 6-year follow-up of individuals at risk of tumour development in either HNPCC or ´´suspected´´ families. There were 374 family members at risk in HNPCC and 468 in ´´suspected´´ families, contributing, respectively, 2,377 and 2,712 persons/year at risk. Thirty-four new tumours developed among HNPCC family members vs. 29.3 expected; however, the occurrence of colorectal cancer in this group was significantly higher than expected, and this excess was particularly evident in the age-groups 41-50 and 51-60. In ´´suspected´´ HNPCC, 38 new tumours developed vs. 24.5 expected; at variance with the other group, besides colorectal neoplasms, lung, liver and brain tumours also occurred significantly in excess. Moreover, the increased risk was uniformly distributed among different age-groups. In conclusion, HNPCC family members are at increased risk of developing colorectal cancer, with an earlier onset than the general population; in contrast, high-risk individuals in ´´suspected´´ HNPCC families seem to be prone to a broader neoplastic spectrum, and risk of tumours does not seem to be limited to any particular age-group.

1993 - Tumour spectrum in hereditary non-polyposis colorectal cancer (HNPCC) and in families with "suspected HNPCC". A population-based study in Northern Italy. Colorectal Cancer Study Group. [Articolo su rivista]
Benatti, Piero; R., Sassatelli; Roncucci, Luca; Pedroni, Monica; R., Fante; C., DI GREGORIO; Losi, Lorena; Gelmini, Roberta; PONZ DE LEON, Maurizio
abstract

Hereditary non-polyposis colorectal cancer (HNPCC or Lynch syndrome) is characterized by the early onset of colorectal neoplasms, frequently localized in the right colon, increased occurrence of multiple primaries, vertical transmission and aggregation of tumours in families in accordance to a Mendelian dominant type of inheritance. The syndrome accounts for approximately 5% of all colorectal cancers. The purpose of the present study was to describe the tumour spectrum and the most relevant clinical features of 28 kindreds with HNPCC, classified according to the guidelines of the international Collaborative Study Group, and of 61 ''suspected'' HNPCC. These families were observed during a 6-year registration of colorectal neoplasms in a health-care district of Northern Italy. Colorectal cancer was by far the most frequent malignancy; gastric cancer was the second. Uterine carcinoma was only slightly more frequent than expected. Lung- and breast-tumour rates were lower than expected. Cancer distribution in the large bowel showed that about two fifths of the tumours developed in the right colon. The occurrence of cancer before the age of SO to 60 was much more frequent in HNPCC. Multiple tumours developed in 25 patients with HNPCC and in 32 with ''suspected'' HNPCC. Pancolonoscopy remains the procedure of choice for surveillance; other examinations, such as gastroscopy, gynaecological investigations, urography and cholangiography, are suggested only to selected families. One of the main features of the study was the inclusion of 61 ''suspected'' HNPCC, a heterogeneous group of families which nonetheless deserves careful follow-up.

1992 - Cell kinetics evaluation of colorectal tumours after in vivo administration of bromodeoxyuridine [Articolo su rivista]
Roncucci, Luca; Pedroni, Monica; A., Scalmati; Ml, Bormioli; R., Sassatelli; R., Fante; Losi, Lorena; C., DI GREGORIO; B., Petocchi; PONZ DE LEON, Maurizio
abstract

Although several biomarkers have been tested, Dukes' (or TNM) stage at diagnosis is still considered the only prognostic factor of clinical relevance in colorectal cancer. Among the various biomarkers, the fraction of cells engaged in DNA synthesis has been extensively investigated as an indicator of tumor aggressiveness. Bromodeoxyuridine (BUdR) is a non-radioactive thymidine analogue which is incorporated into DNA during the S-phase of cycling cells. In order to evaluate the relationships between cell kinetics and morphologic variables, 500 mg of BUdR were given i.v. to patients with colorectal cancer prior to surgery. After operation, a large tumor sample was taken and processed for immunohistochemical detection of BUdR-labeled cells in various regions of the neoplasm and in normal colorectal mucosa. Smaller superficial tumor specimens were also incubated with H-3-thymidine (H-3-TdR) for the autoradiographic identification of labeled cells. In the 43 evaluable tumors, the overall BUdR labeling index (BLI, percent of labeled cells) was significantly higher in carcinoma (20.30 +/- 0.86%, SEM) than in normal colonic mucosa (6.51 +/- 0.49%). BLIs in central and peripheral regions of carcinoma were closely correlated (r = 0.48, p = 0.003). In 21 neoplasms a high correlation between overall BUdR and H-3-TdR labeling index in the same tumor was observed (r = 0.57, p = 0.007). No evident association between overall BU and clinical or morphologic parameters of the tumor was seen, including number of capillaries and ras-p21 protein expression. We conclude that BUdR immunostaining after in vivo administration of BUdR is a simple method for studying cell kinetics in various regions of colorectal cancer. BUdR labeling data are comparable to those obtained with in vitro incorporation of H-3-TdR.

1991 - Biological evaluation of colorectal neoplasia [Articolo su rivista]
PONZ DE LEON, Maurizio; A., Scalmati; Pedroni, Monica; Roncucci, Luca; C., Di Gregorio; R., Fante
abstract

Not applicable

1991 - Cell kinetics parameters in human tumors in vivo or determined using bromodeoxyuridine incorporation and flow cytometry [Articolo su rivista]
E., Geido; W., Giaretti; A., Di Vinci; R., Corvò; Vivitale, ; G., Margarino; Roncucci, Luca; Pedroni, Monica; PONZ DE LEON, Maurizio
abstract

Not applicable