Taking an insight into the cytogenic and epigenetic techniques, the current study aims at characterizing Rionero in Vulture IRCCS CROB patients, diagnosed with Chronic Lymphocytic Leukemia (CLL), an extremely heterogenous and hematological neoplasm. The patients under study, analyzed over a 5 year range (2016-2021), were first characterized clinically (complete blood count), through the analysis of the immunophenotype and then, genetically with the mutational analysis of the TP53 tumor suppressor gene. Interphase FISH analysis was performed to identify cytogenetic alterations and stratify patients into different risk ranges. Our study, then, followed up by determining the methylation patterns of patients under study to investigate the epigenetic-genetic cross talk and investigate a possible trend of differentially methylated genes (DMGs) among patients with different risk stratification. Through a gene set enrichment analysis, the alteration of some pathways emerged, already extensively analyzed in previous studies, and which appeared to be involved in the pathogenesis of CLL. In addition, chromosomal positions with differentially methylated genes have been identified among patients with different prognoses. In particular, our study only shows the results related to the dataset of hypermethylated genes. The limit of the current study consists of the small number of cases and the lack of information on the patients’ conditions (ex. expression of lipid lipoprotein (LPL)), as well as different kinds of analyses, such as the mutational status of NOTCH1, which, currently, are not carried out at our facility. The results obtained have shown the need to investigate further the influence from events that can affect the gene expression of the chr17p13 region, TP53 locus, commonly deleted in CLL and responsible for an adverse prognosis.
Il presente studio è nato con l’intento di caratterizzare da un punto di vista citogenetico ed epigenetico pazienti dell’IRCCS CROB di Rionero in Vulture con diagnosi di leucemia linfatica cronica (LLC), una neoplasia ematologica estremamente eterogenea. I pazienti selezionati nell’arco di 5 anni (range 2016-2021) sono stati inizialmente caratterizzati da un punto di vista clinico (emocromo), attraverso l’analisi dell’immunofenotipo e da un punto di vista genetico con l’analisi mutazionale del gene oncosoppressore TP53. È stata eseguita l’analisi FISH in interfase per individuare le alterazioni citogenetiche e stratificare i pazienti in diverse fasce di rischio. Abbiamo proseguito determinando i profili di metilazione dei soggetti in studio per approfondire il cross talk epigenetico-genetico ed indagare su una possibile tendenza di geni differenzialmente metilati (DMGs) tra pazienti con diversa stratificazione di rischio. Con un’analisi dell'arricchimento del set di geni differenzialmente metilati è emersa l’alterazione di alcuni pathway, abbondantemente descritti in letteratura per essere coinvolti nella patogenesi della LLC; inoltre, sono state identificate posizioni cromosomiche con geni differenzialmente metilati tra pazienti con diversa prognosi. In particolare, in questo lavoro di tesi sono riportati solo i risultati relativi al dataset di geni ipermetilati. Il limite del seguente studio è dato dalla casistica poco numerosa e dalla mancanza di informazioni sullo status dei pazienti (es. espressione della lipoproteina lipidica (LPL)), nonché di differenti analisi, quali ad esempio lo stato mutazionale di NOTCH1, che al momento non vengono effettuate presso l’istituto. Dai risultati ottenuti emerge la necessità di approfondire la suscettibilità ad eventi che possono influenzare l’espressione genica della regione chr17p13, locus di TP53, comunemente deleto nella LLC e responsabile di prognosi sfavorevole.
CARATTERIZZAZIONE CITOGENETICA E DEL PROFILO DI METILAZIONE DI PAZIENTI CON LEUCEMIA LINFATICA CRONICA
OUAZRI, SOFIA
2020/2021
Abstract
Taking an insight into the cytogenic and epigenetic techniques, the current study aims at characterizing Rionero in Vulture IRCCS CROB patients, diagnosed with Chronic Lymphocytic Leukemia (CLL), an extremely heterogenous and hematological neoplasm. The patients under study, analyzed over a 5 year range (2016-2021), were first characterized clinically (complete blood count), through the analysis of the immunophenotype and then, genetically with the mutational analysis of the TP53 tumor suppressor gene. Interphase FISH analysis was performed to identify cytogenetic alterations and stratify patients into different risk ranges. Our study, then, followed up by determining the methylation patterns of patients under study to investigate the epigenetic-genetic cross talk and investigate a possible trend of differentially methylated genes (DMGs) among patients with different risk stratification. Through a gene set enrichment analysis, the alteration of some pathways emerged, already extensively analyzed in previous studies, and which appeared to be involved in the pathogenesis of CLL. In addition, chromosomal positions with differentially methylated genes have been identified among patients with different prognoses. In particular, our study only shows the results related to the dataset of hypermethylated genes. The limit of the current study consists of the small number of cases and the lack of information on the patients’ conditions (ex. expression of lipid lipoprotein (LPL)), as well as different kinds of analyses, such as the mutational status of NOTCH1, which, currently, are not carried out at our facility. The results obtained have shown the need to investigate further the influence from events that can affect the gene expression of the chr17p13 region, TP53 locus, commonly deleted in CLL and responsible for an adverse prognosis.È consentito all'utente scaricare e condividere i documenti disponibili a testo pieno in UNITESI UNIPV nel rispetto della licenza Creative Commons del tipo CC BY NC ND.
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https://hdl.handle.net/20.500.14239/14706