TY - JOUR T1 - Four-miRNA Signature to Identify Asbestos-Related Lung Malignancies. JF - Cancer Epidemiol Biomarkers Prev Y1 - 2019 A1 - Santarelli, Lory A1 - Gaetani, Simona A1 - Monaco, Federica A1 - Bracci, Massimo A1 - Valentino, Matteo A1 - Amati, Monica A1 - Rubini, Corrado A1 - Sabbatini, Armando A1 - Pasquini, Ernesto A1 - Zanotta, Nunzia A1 - Comar, Manola A1 - Neuzil, Jiri A1 - Tomasetti, Marco A1 - Bovenzi, Massimo AB -

BACKGROUND: Altered miRNA expression is an early event upon exposure to occupational/environmental carcinogens; thus, identification of a novel asbestos-related profile of miRNAs able to distinguish asbestos-induced cancer from cancer with different etiology can be useful for diagnosis. We therefore performed a study to identify miRNAs associated with asbestos-induced malignancies.

METHODS: Four groups of patients were included in the study, including patients with asbestos-related (NSCLC) and asbestos-unrelated non-small cell lung cancer (NSCLC) or with malignant pleural mesothelioma (MPM), and disease-free subjects (CTRL). The selected miRNAs were evaluated in asbestos-exposed population.

RESULTS: Four serum miRNAs, that is miR-126, miR-205, miR-222, and miR-520g, were found to be implicated in asbestos-related malignant diseases. Notably, increased expression of miR-126 and miR-222 were found in asbestos-exposed subjects, and both miRNAs are involved in major pathways linked to cancer development. Epigenetic changes and cancer-stroma cross-talk could induce repression of miR-126 to facilitate tumor formation, angiogenesis, and invasion.

CONCLUSIONS: This study indicates that miRNAs are potentially involved in asbestos-related malignancies, and their expression outlines mechanism(s) whereby miRNAs may be involved in an asbestos-induced pathogenesis.

IMPACT: The discovery of a miRNA panel for asbestos-related malignancies would impact on occupational compensation and may be utilized for screening asbestos-exposed populations.

VL - 28 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/30257964?dopt=Abstract ER - TY - JOUR T1 - Combined circulating epigenetic markers to improve mesothelin performance in the diagnosis of malignant mesothelioma. JF - Lung Cancer Y1 - 2015 A1 - Santarelli, Lory A1 - Staffolani, Sara A1 - Strafella, Elisabetta A1 - Nocchi, Linda A1 - Manzella, Nicola A1 - Grossi, Paola A1 - Bracci, Massimo A1 - Pignotti, Elettra A1 - Alleva, Renata A1 - Borghi, Battista A1 - Pompili, Cecilia A1 - Sabbatini, Armando A1 - Rubini, Corrado A1 - Zuccatosta, Lina A1 - Bichisecchi, Elisabetta A1 - Valentino, Matteo A1 - Horwood, Keith A1 - Comar, Manola A1 - Bovenzi, Massimo A1 - Dong, Lan-Feng A1 - Neuzil, Jiri A1 - Amati, Monica A1 - Tomasetti, Marco AB -

OBJECTIVES: Malignant mesothelioma (MM) is a highly aggressive tumor with poor prognosis. A major challenge is the development and application of early and highly reliable diagnostic marker(s). Serum biomarkers, such as 'soluble mesothelin-related proteins' (SMRPs), is the most studied and frequently used in MM. However, the low sensitivity of SMRPs for early MM limits its value; therefore, additional biomarkers are required. In this study, two epigenetically regulated markers in MM (microRNA-126, miR-126, and methylated thrombomodulin promoter, Met-TM) were combined with SMRPs and evaluated as a potential strategy to detect MM at an early stage.

MATERIALS AND METHODS: A total of 188 subjects, including 45 MM patients, 99 asbestos-exposed subjects, and 44 healthy controls were prospectively enrolled, serum samples collected, and serum levels of SMRPs, miR-126 and Met-TM evaluated. Logistic regression analysis was performed to evaluate the diagnostic value of the three biomarkers. Using this approach, the performance of the '3-biomarker classifier' was tested by calculating the overall probability score of the MM and control samples, respectively, and the ROC curve was generated.

RESULTS AND CONCLUSION: The combination of the three biomarkers was the best predictor to differentiate MM patients from asbestos-exposed subjects and healthy controls. The accuracy and cancer specificity was confirmed in a second validation cohort and lung cancer population. We propose that the combination of the two epigenetic biomarkers with SMRPs as a diagnosis for early MM overcomes the limitations of using SMRPs alone.

U1 - http://www.ncbi.nlm.nih.gov/pubmed/26431916?dopt=Abstract ER - TY - JOUR T1 - Immunologic evidence of a strong association between non-Hodgkin lymphoma and simian virus 40. JF - Cancer Y1 - 2015 A1 - Tognon, Mauro A1 - Luppi, Mario A1 - Corallini, Alfredo A1 - Taronna, Angelo A1 - Barozzi, Patrizia A1 - Rotondo, John Charles A1 - Comar, Manola A1 - Casali, Maria Vittoria A1 - Bovenzi, Massimo A1 - D'Agostino, Antonio A1 - Vinante, Fabrizio A1 - Rigo, Antonella A1 - Ferrarini, Isacco A1 - Barbanti-Brodano, Giuseppe A1 - Martini, Fernanda A1 - Mazzoni, Elisa KW - Adult KW - Aged KW - Antibodies, Viral KW - Capsid Proteins KW - Enzyme-Linked Immunosorbent Assay KW - Female KW - Humans KW - Lymphoma, Non-Hodgkin KW - Male KW - Middle Aged KW - Polyomavirus Infections KW - Seroepidemiologic Studies KW - Simian virus 40 KW - Tumor Virus Infections AB -

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

METHODS: In this study, an indirect enzyme-linked immunosorbent assay with 2 synthetic peptides that mimic SV40 antigens of viral capsid proteins 1 to 3 was employed to detect specific antibodies against SV40. Serum samples were taken from 2 distinct cohorts of NHL-affected patients (NHL1 [n = 89] and NHL2 [n = 61]) along with controls represented by oncologic patients affected by breast cancer (BC; n = 78) and undifferentiated nasopharyngeal carcinoma (UNPC; n = 64) and 3 different cohorts of healthy subjects (HSs; HS1 [n = 130], HS2 [n = 83], and HS3 [n = 87]).

RESULTS: Immunologic data indicated that in serum samples from NHL patients, antibodies against SV40 mimotopes were detectable with a prevalence of 40% in NHL1 patients and with a prevalence of 43% in NHL2 patients. In HSs of the same median age as NHL patients, the prevalence was 16% for the HS1 group (57 years) and 14% for the HS2 group (65 years). The difference was statistically significant (P < .0001 and P < .001). Interestingly, the difference between NHL1/NHL2 patients and BC patients (40%/43% vs 15%, P < .001) and between NHL1/NHL2 patients and UNPC patients (40%/43% vs 25%, P < .05) was significant.

CONCLUSIONS: Our data indicate a strong association between NHL and SV40 and thus a need for innovative therapeutic approaches for this hematologic malignancy.

VL - 121 IS - 15 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25877010?dopt=Abstract ER - TY - JOUR T1 - Increased levels of C-C chemokine RANTES in asbestos exposed workers and in malignant mesothelioma patients from an hyperendemic area. JF - PLoS One Y1 - 2014 A1 - Comar, Manola A1 - Zanotta, Nunzia A1 - Bonotti, Alessandra A1 - Tognon, Mauro A1 - Negro, Corrado A1 - Cristaudo, Alfonso A1 - Bovenzi, Massimo KW - Aged KW - Aged, 80 and over KW - Asbestos KW - Biomarkers, Tumor KW - Case-Control Studies KW - Chemokine CCL5 KW - Cytokines KW - Endemic Diseases KW - Gene Expression KW - Gene Expression Profiling KW - Humans KW - Lung Neoplasms KW - Mesothelioma KW - Middle Aged KW - Occupational Exposure KW - Simian virus 40 KW - Viral Proteins AB -

BACKGROUND: Asbestos-induced mesothelial inflammatory processes are thought to be the basic mechanisms underlying Malignant Mesothelioma (MM) development. Detection of MM often occurs at late stage due to the long and unpredictable latent period and the low incidence in asbestos exposed individuals. The aim of this study was to investigate early immunological biomarkers to characterize the prognostic profile of a possible asbestos-induced disease, in subjects from a MM hyperendemic area.

METHODS: The Luminex Multiplex Panel Technology was used for the simultaneous measurement of serum levels of a large panel of 47 analytes, including cytokines and growth factors, from workers previously exposed to asbestos (Asb-workers), asbestos-induced MM patients and healthy subjects. In addition, to explore the influence on serum cytokines profile exerted by SV40 infection, a cofactor in MM development, a quantitative real time PCR was performed for sequences detection in the N-terminal and intronic regions of the SV40 Tag gene. Statistical analysis was done by means of the Mann-Whitney test and the Kruskall-Wallis test for variance analysis.

RESULTS: A variety of 25 cytokines linked to pulmonary inflammation and tumor development were found significantly associated with Asb-workers and MM patients compared with healthy controls. A specific pattern of cytokines were found highly expressed in Asb-workers: IFN-alpha (p<0.05), EOTAXIN (p<0.01), RANTES (p<0.001), and in MM patients: IL-12(p40), IL-3, IL-1 alpha, MCP-3, beta-NGF, TNF-beta, RANTES (p<0.001). Notably, the chemokine RANTES measured the highest serum level showing an increased gradient of concentration from healthy subjects to Asb-workers and MM patients (p<0.001), independently of SV40 infection.

CONCLUSION: This study shows that, in subjects from an hyperendemic area for MM, the C-C chemokine RANTES is associated with the exposure to asbestos fibres. If validated in larger samples, this factor could have the potential to be a critical biomarker for MM prognosis as recently reported for breast tumor.

VL - 9 IS - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25162674?dopt=Abstract ER - TY - JOUR T1 - Asbestos and SV40 in malignant pleural mesothelioma from a hyperendemic area of north-eastern Italy. JF - Tumori Y1 - 2012 A1 - Comar, Manola A1 - Zanotta, Nunzia A1 - Pesel, Giuliano A1 - Visconti, Patrizia A1 - Maestri, Iva A1 - Rinaldi, Rosa A1 - Crovella, Sergio A1 - Cortale, Maurizio A1 - De Zotti, Renata A1 - Bovenzi, Massimo KW - Adult KW - Aged KW - Asbestos KW - Carcinogens KW - Disease Susceptibility KW - DNA, Viral KW - Endemic Diseases KW - Female KW - Humans KW - Italy KW - Male KW - Mesothelioma KW - Middle Aged KW - Pleural Neoplasms KW - Polyomavirus Infections KW - Real-Time Polymerase Chain Reaction KW - Risk Factors KW - Simian virus 40 KW - Tumor Virus Infections KW - Viral Load AB -

AIMS AND BACKGROUND: Malignant mesothelioma is a fatal cancer of increasing incidence in north-eastern Italy. Together with asbestos, the polyomavirus SV40 was hypothesized to contribute to the onset of malignant mesothelioma. To investigate the putative role of SV40 in the individual susceptibility to asbestos-induced malignant mesothelioma, we conducted a molecular epidemiological study on a series of malignant mesothelioma patients from an area in north-eastern Italy hyperendemic for malignant pleural mesothelioma.

METHODS AND STUDY DESIGN: We collected 63 mesothelioma samples from incidence cases of patients diagnosed with malignant pleural mesothelioma in the period 2009-2010. DNA was extracted from patients' tissue biopsies using the BioRobot EZ1 Qiagen workstation. SV40 sequence detection and quantification was performed by specific real time PCR. The 74.6% of the 63 enrolled patients had a history of asbestos exposure. The epithelioid histotype was more prevalent in males (64.0%) and the mixed in females (61.5%) who showed significantly higher cancer co-morbidity (46.1% vs 12%, P = 0.005). SV40 was detected in 22% of MM tumors, with a low viral load. In SV40-positive patients, a threefold increased risk of asbestos exposure was observed, more evident in females (OR 4.32) than in males (OR 1.20).

CONCLUSIONS: Our findings indicate that a high prevalence of SV40 was present in malignant mesothelioma incident cases from an area hyperendemic for malignant mesothelioma in north-eastern Italy. Although asbestos is considered the main risk factor in malignant mesothelioma onset, a role for SV40 could be hypothesized.

VL - 98 IS - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22677986?dopt=Abstract ER - TY - JOUR T1 - High prevalence of serum antibodies reacting with simian virus 40 capsid protein mimotopes in patients affected by malignant pleural mesothelioma. JF - Proc Natl Acad Sci U S A Y1 - 2012 A1 - Mazzoni, Elisa A1 - Corallini, Alfredo A1 - Cristaudo, Alfonso A1 - Taronna, Angelo A1 - Tassi, Gianfranco A1 - Manfrini, Marco A1 - Comar, Manola A1 - Bovenzi, Massimo A1 - Guaschino, Roberto A1 - Vaniglia, Francesca A1 - Magnani, Corrado A1 - Casali, Ferruccio A1 - Rezza, Giovanni A1 - Barbanti-Brodano, Giuseppe A1 - Martini, Fernanda A1 - Tognon, Mauro G KW - Amino Acid Sequence KW - Antibodies, Viral KW - Capsid Proteins KW - Enzyme-Linked Immunosorbent Assay KW - Female KW - Humans KW - Male KW - Mesothelioma KW - Molecular Sequence Data KW - Pleural Neoplasms KW - Pregnancy KW - Simian virus 40 AB -

Human malignant pleural mesothelioma (MPM) is considered a rare tumor, but recent estimations indicate that one-quarter million people will die of this neoplasm in Europe in the next three decades. The mineral asbestos is considered the main causative agent of this neoplasm. MPM is largely unresponsive to conventional chemotherapy/radiotherapy. In addition to asbestos exposure, genetic predisposition to asbestos carcinogenesis and to simian virus (SV)40 infection has also been suggested. SV40 is a DNA tumor virus found in some studies to be associated at high prevalence with MPM. SV40 sequences have also been detected, although at a lower prevalence than in MPM, in blood specimens from healthy donors. However, some studies have failed to reveal SV40 footprints in MPM and its association with this neoplasm. These conflicting results indicate the need for further investigations with new approaches. We report on the presence of antibodies in serum samples from patients affected by MPM that specifically react with two different SV40 mimotopes. The two SV40 peptides used in indirect ELISAs correspond to viral capsid proteins. ELISA with the two SV40 mimotopes gave overlapping results. Our data indicate that in serum samples from MPM-affected patients (n = 97), the prevalence of antibodies against SV40 viral capsid protein antigens is significantly higher (26%, P = 0.043) than in the control group (15%) represented by healthy subjects (n = 168) with the same median age (66 y) and sex. Our results suggest that SV40 is associated with a subset of MPM and circulates in humans.

VL - 109 IS - 44 U1 - http://www.ncbi.nlm.nih.gov/pubmed/23071320?dopt=Abstract ER - TY - JOUR T1 - NLRP1 polymorphisms in patients with asbestos-associated mesothelioma. JF - Infect Agent Cancer Y1 - 2012 A1 - Girardelli, Martina A1 - Maestri, Iva A1 - Rinaldi, Rosa R A1 - Tognon, Mauro A1 - Boldorini, Renzo A1 - Bovenzi, Massimo A1 - Crovella, Sergio A1 - Comar, Manola AB -

UNLABELLED:

BACKGROUND: An increasing incidence of malignant mesothelioma (MM) cases in patients with low levels of asbestos exposure suggests the interference of alternative cofactors. SV40 infection was detected, as co-morbidity factor, only in 22% of asbestos-MM patients from a North-Eastern Italy area. An additional mechanism of injury related to asbestos exposure in MM development has been recently associated to inflammatory responses, principally driven by interleukin (IL)-1 beta (ß) activated within the inflammasome complex.NLRP3 inflammosome has been described as the intracellular sensor for asbestos able to induce inflammasome activation and IL-1ß secretion while NLRP1 is expressed in lung epithelial cells and alveolar macrophages and contributes to the immune response and to survival/apoptosis balance. This study proposes to evaluate the impact of known NLRP3 and NLRP1 polymorphisms in the individual susceptibility to asbestos-induced mesothelioma in subjects from a hyperendemic area for MM.

METHODS: 134 Italian patients with diagnosis of mesothelioma due (MMAE, n=69) or not (MMAF, n=65) to asbestos, 256 healthy Italian blood donors and 101 Italian healthy subjects exposed to asbestos (HCAE) were genotyped for NLRP1 (rs2670660 and rs12150220) and NLRP3 (rs35829419 and rs10754558) polymorphisms.

RESULTS: While NLRP3 SNPs were not associated to mesothelioma, the NLRP1 rs12150220 allele T was significantly more frequent in MMAE (0.55) than in HCAE (0.41) (p=0.011; OR=1.79) suggesting a predisponent effect of this allele on the development of mesothelioma. This effect was amplified when the NLRP1 rs2670660 allele was combined with the NLRP1 rs12150220 allele (p=0.004; OR=0.52).

CONCLUSION: Although NLRP3 SNPs was not involved in mesothelioma predisposition, these data proposed NLRP1 as a novel factor possibly involved in the development of mesothelioma.

VL - 7 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/23031505?dopt=Abstract ER - TY - JOUR T1 - The significance of mannose-binding lectin gene polymorphisms on the risk of BK virus coinfection in women with human papillomavirus-positive cervical lesions. JF - Hum Immunol Y1 - 2011 A1 - Comar, Manola A1 - Segat, Ludovica A1 - Crovella, Sergio A1 - Bovenzi, Massimo A1 - Cortini, Enzo A1 - Tognon, Mauro KW - Adult KW - Aged KW - Alleles KW - BK Virus KW - Cervical Intraepithelial Neoplasia KW - Cervix Uteri KW - DNA Fingerprinting KW - DNA, Viral KW - Female KW - Gene Frequency KW - Genetic Association Studies KW - Genetic Predisposition to Disease KW - Genotype KW - Human papillomavirus 16 KW - Humans KW - Italy KW - Mannose-Binding Lectin KW - Odds Ratio KW - Papillomavirus Infections KW - Polymorphism, Genetic KW - Polyomavirus Infections KW - Risk KW - Viral Load AB -

The simultaneous detection of oncogenic human papillomavirus (HPV) and BK virus (BKV) has been recently reported in cervical cancers, suggesting that these viruses may act together in the process of cell transformation; host genetic polymorphisms may also influence virus persistence/reactivation. To disclose a possible role of the gene encoding for the mannose-binding lectin, MBL2, in susceptibility to BKV infection, we analyzed functional polymorphisms in the first exon of MBL2 in women stratified for the presence/absence of BKV and affected by different grades of HPV-induced cervical precancerous lesions. All BKV-positive samples were also HPV positive (HPV 16), and all presented with high-grade squamous intraepithelial lesions. The MBL2 A allele was significantly more frequent in BKV-negative patients than in BKV-positive patients. These data indicate a possible role for the A allele in conferring protection to BKV infection in high-risk HPV-positive women (odds ratio 0.40, 95% confidence interval 0.20-0.85, p = 0.01).

VL - 72 IS - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21536088?dopt=Abstract ER - TY - JOUR T1 - JCV/BKV and SV40 viral load in lymphoid tissues of young immunocompetent children from an area of north-east Italy. JF - J Med Virol Y1 - 2010 A1 - Comar, Manola A1 - Zanotta, Nunzia A1 - Bovenzi, Massimo A1 - Campello, Cesare KW - Adenoids KW - BK Virus KW - Carrier State KW - Child KW - Child, Preschool KW - Female KW - Herpesvirus 6, Human KW - Humans KW - Immunocompetence KW - Italy KW - JC Virus KW - Male KW - Neutrophils KW - Palatine Tonsil KW - Polyomavirus Infections KW - Simian virus 40 KW - Viral Load KW - Virus Latency AB -

Polyomavirus infection occurring during childhood is followed by a lifelong latency in immunocompetent subjects. The major site of polyomavirus persistence are the uroepithelial cells which leads to oral transmission. It has recently been hypothesized that tonsils could be a possible reservoir. The role of tonsil, adenoid, and peripheral blood mononuclear cells (PBMCs) as possible sites of JCV, BKV, and SV40 latency in young healthy children was assessed. Two hundred fifteen fresh specimens, including 57 tonsil, 80 adenoid, and 78 PBMC samples from 80 immunocompetent children (mean age 4.8 years) were analyzed to determine the viral load by quantitative real-time PCR. The human herpes virus 6 (HHV-6) was tested as a lymphotropic reference virus. Polyomavirus was detected in 5/80 (6.2%) children while HHV-6 infection affected 27/80 children (33.7%) (P < 0.001). SV40 was detected in one adenoid sample, while footprints of BKV were found in one adenoid and three tonsil samples. JCV was never found in all samples. Polyomavirus sequences were not detected in the 78 blood samples. One adenoid and two tonsils from three children (1.4%) were positive for both polyomavirus and HHV-6. Infections were characterized by low replication rates ranging typically from 1 x 10e(2)/5.5 x 10e(4) to 6.8 x 10e(3)/8.5 x 10e(4) viral copies/number of cells. In conclusion, tonsils and adenoids of children could effectively harbor BKV and SV40, although only very few cells proved to be infected. Nevertheless, the low prevalence of polyomavirus, in comparison with the lymphotropic HHV-6, suggests that these tissues are unlikely to be the preferred site of polyomavirus latency, at least in younger children.

VL - 82 IS - 7 U1 - http://www.ncbi.nlm.nih.gov/pubmed/20513090?dopt=Abstract ER -