Supervisors info:
Μαριάννα Αντωνέλου, Επίκουρη Καθηγήτρια, Τμήμα Βιολογίας, ΕΚΠΑ
Summary:
Objective: Μyelodysplastic/myeloproliferative neoplasms are myeloid disorders characterized by both dysplastic and proliferative features. Molecular lesions in genes involved in cell signaling, epigenetic regulation (TET2 and IDH1 / 2), and RNA splicing (SRSF2, SF3B1, U2AF1) are often detected in a large proportion of patients. The latter are not exclusive, and are usually detected with mutation in other genes, i.e. DNMT3, JAK2, ASXL1 and TET2. The purpose of this thesis was to estimate the gene methylation by analyzing the LINE-1 retrotransposon elements that are dispersed in the genome utilizing α post-Real-Time PCR HRMA technique in samples of patients with MDS/MPN.
Methods: In this study, 60 patients were included based on WHO 2016 criteria for MDS/MPN and according to their molecular profile, 15 patients were selected for methylation analysis. To determine the methylation status of the LINE-1, a standard curve was prepared using 6 DNA samples from healthy donors. For this purpose, the MS-HRMA method was used and the MSP method was used to verify the data.
Results: The LINE-1 retrotransposon elements of MDS/MPN patients showed statistically significant (p-value = 0.0071), increased methylation levels (88.47%) in samples from patients with MDS/MPN, compared to the methylation levels iDNA samples from healthy donors (85.81%). Additionally, in MDS/MPN patients with a mutation in the DNMT3A gene, the LINE-1 transposons methylation status significantly reduced (86.77%;p-value = 0.0322), in comparison to methylation levels in patients with mutations in other genes (88.73%). The results were verified by the MSP method.
Conclusions: In this study, we showed a statistically significant difference in the methylation levels of LINE-1 retrotransposon elements (p-value = 0.0036) in patients with MDS/MPN compared to the normal population. Further analysis should be performed in larger cohorts of patients to confirm these results.
