Occupational or environmental chronic exposure to pesticides is associated with the development of various chronic degenerative diseases, including eye tumors. As is known, some of these pesticides are cataloged as certain carcinogens for humans as they are capable of inducing mutations and direct damage to DNA or cellular structures, thus determining the onset of a pathological state. More recently, several studies have shown how numerous classes of pesticides are able to induce not only direct molecular or cellular damage, but also indirect damage by inducing epigenetic modifications, such as DNA methylation and alteration of the expression levels of specific miRNAs. On these bases, the purpose of this work was to analyze the bioinformatics data present in the literature related to the epigenetic modifications induced by pesticides as well as the alteration of miRNA expression levels observed in uveal melanoma in order to identify a panel of genetic and epigenetic markers to be used to monitor the health status of individuals occupationally exposed to these contaminants and thus predict the risk of uveal melanoma development according to the exposure patterns to specific pesticides. In particular, using different computational analyses, various datasets of gene expression, DNA methylation and expression of miRNAs were analyzed taking into account experiments performed on in vitro experimental studies, animal models and observational studies on individuals exposed to pesticides. Through the analysis and integration of these datasets it was possible to identify genetic markers, miRNAs and methylation hotspots whose alteration is indicative of exposure to pesticides and an increased risk of the onset of chronic degenerative diseases. In parallel, the computational analyses performed on uveal melanoma datasets allowed the identification of miRNA associated with the development and progression of this tumor. Through the integration of the bioinformatics data obtained for pesticide exposure and the development of uveal melanoma it was possible to select a panel of miRNAs associated with uveal melanoma development and associated with pesticide exposure. Such miRNAs were also validated in a cohort of uveal melanoma patients where the expression levels of miRNAs were assessed on liquid biopsy samples and FFPE tissues obtained from uveal melanoma patients and normal controls. Through these analyses it was demonstrated the involvement of miRNA in uveal melanoma development as well as the detrimental role of pesticide exposure able to induce epigenetics alterations predisposing tumor development. The results obtained in this study pave the way to the development of novel diagnostic and prognostic strategies for the prediction of uveal melanoma development in individuals professionally exposed to pesticides through the analysis of the expression levels of miRNAs as indicators of pesticide exposure and epigenetic damages.
L'esposizione cronica professionale o ambientale ai pesticidi è associata allo sviluppo di varie malattie degenerative croniche, compresi i tumori oculari. Come è noto, alcuni di questi pesticidi sono catalogati come cancerogeni per l'uomo in quanto sono in grado di indurre mutazioni e danni diretti al DNA o alle strutture cellulari, determinando così l'insorgere di uno stato patologico. Più recentemente, diversi studi hanno mostrato come numerose classi di pesticidi siano in grado di indurre non solo danni molecolari o cellulari diretti, ma anche danni indiretti inducendo modificazioni epigenetiche, come la metilazione del DNA e l'alterazione dei livelli di espressione di specifici miRNA. Su queste basi, lo scopo di questo lavoro è stato quello di analizzare i dati bioinformatici presenti in letteratura relativi alle modificazioni epigenetiche indotte dai pesticidi nonché l'alterazione dei livelli di espressione dei miRNA osservati nel melanoma uveale al fine di identificare un pannello di fattori genetici ed epigenetici marcatori da utilizzare per monitorare lo stato di salute degli individui professionalmente esposti a questi contaminanti e quindi prevedere il rischio di sviluppo di melanoma uveale in base ai modelli di esposizione a specifici pesticidi. In particolare, utilizzando diverse analisi computazionali, sono stati analizzati vari dataset di espressione genica, metilazione del DNA ed espressione di miRNA tenendo conto di esperimenti condotti su studi sperimentali in vitro, modelli animali e studi osservazionali su individui esposti a pesticidi. Attraverso l'analisi e l'integrazione di questi dataset è stato possibile identificare marcatori genetici, miRNA e hotspot di metilazione la cui alterazione è indicativa dell'esposizione ai pesticidi e di un aumentato rischio di insorgenza di malattie croniche degenerative. Parallelamente, le analisi computazionali eseguite sui dataset del melanoma uveale hanno permesso di identificare i miRNA associati allo sviluppo e alla progressione di questo tumore. Attraverso l'integrazione dei dati bioinformatici ottenuti per l'esposizione ai pesticidi e lo sviluppo del melanoma uveale è stato possibile selezionare un pannello di miRNA associati allo sviluppo del melanoma uveale e associati all'esposizione ai pesticidi. Tali miRNA sono stati anche convalidati in una coorte di pazienti con melanoma uveale in cui i livelli di espressione di miRNA sono stati valutati su campioni di biopsia liquida e tessuti FFPE ottenuti da pazienti con melanoma uveale e controlli normali. Attraverso queste analisi è stato dimostrato il coinvolgimento dei miRNA nello sviluppo del melanoma uveale nonché il ruolo dannoso dell'esposizione a pesticidi in grado di indurre alterazioni epigenetiche che predispongono allo sviluppo del tumore. I risultati ottenuti in questo studio aprono la strada allo sviluppo di nuove strategie diagnostiche e prognostiche per la previsione dello sviluppo del melanoma uveale in individui professionalmente esposti ai pesticidi attraverso l'analisi dei livelli di espressione dei miRNA come indicatori di esposizione ai pesticidi e danni epigenetici.
Identificazione delle alterazioni epigenetiche indotte dall'esposizione ai pesticidi e validazione del potenziale diagnostico e prognostico dei microRNA nel melanoma uveale / D'Aquino, Manuela. - (2021 Nov 25).
Identificazione delle alterazioni epigenetiche indotte dall'esposizione ai pesticidi e validazione del potenziale diagnostico e prognostico dei microRNA nel melanoma uveale
D'AQUINO, MANUELA
2021-11-25
Abstract
Occupational or environmental chronic exposure to pesticides is associated with the development of various chronic degenerative diseases, including eye tumors. As is known, some of these pesticides are cataloged as certain carcinogens for humans as they are capable of inducing mutations and direct damage to DNA or cellular structures, thus determining the onset of a pathological state. More recently, several studies have shown how numerous classes of pesticides are able to induce not only direct molecular or cellular damage, but also indirect damage by inducing epigenetic modifications, such as DNA methylation and alteration of the expression levels of specific miRNAs. On these bases, the purpose of this work was to analyze the bioinformatics data present in the literature related to the epigenetic modifications induced by pesticides as well as the alteration of miRNA expression levels observed in uveal melanoma in order to identify a panel of genetic and epigenetic markers to be used to monitor the health status of individuals occupationally exposed to these contaminants and thus predict the risk of uveal melanoma development according to the exposure patterns to specific pesticides. In particular, using different computational analyses, various datasets of gene expression, DNA methylation and expression of miRNAs were analyzed taking into account experiments performed on in vitro experimental studies, animal models and observational studies on individuals exposed to pesticides. Through the analysis and integration of these datasets it was possible to identify genetic markers, miRNAs and methylation hotspots whose alteration is indicative of exposure to pesticides and an increased risk of the onset of chronic degenerative diseases. In parallel, the computational analyses performed on uveal melanoma datasets allowed the identification of miRNA associated with the development and progression of this tumor. Through the integration of the bioinformatics data obtained for pesticide exposure and the development of uveal melanoma it was possible to select a panel of miRNAs associated with uveal melanoma development and associated with pesticide exposure. Such miRNAs were also validated in a cohort of uveal melanoma patients where the expression levels of miRNAs were assessed on liquid biopsy samples and FFPE tissues obtained from uveal melanoma patients and normal controls. Through these analyses it was demonstrated the involvement of miRNA in uveal melanoma development as well as the detrimental role of pesticide exposure able to induce epigenetics alterations predisposing tumor development. The results obtained in this study pave the way to the development of novel diagnostic and prognostic strategies for the prediction of uveal melanoma development in individuals professionally exposed to pesticides through the analysis of the expression levels of miRNAs as indicators of pesticide exposure and epigenetic damages.File | Dimensione | Formato | |
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