Testicular germ cell tumor (TGCT) patients are frequently exposed to cytotoxic therapies that may induce epigenetic alterations in sperm DNA, potentially affecting reproductive outcomes and embryonic development [1,2]. This study evaluated the methylation status of four genes involved in genomic imprinting and embryogenesis, PEG1/MEST, IGF2, H19, and BCL2, in sperm DNA from TGCT patients before and after chemotherapy [3]. Twenty-four TGCT patients were included in a longitudinal cohort providing semen samples before treatment (T0) and two years after therapy completion (T2). An additional cross-sectional cohort included 31 post-treatment patients and 10 fertile normozoospermic controls. Sperm DNA underwent bisulfite conversion followed by pyrosequencing analysis of CpG site within the promoter regions of the selected genes. In the longitudinal analysis, no statistically significant differences in methylation levels were observed for any of the analyzed genes. Similarly, stratification according to chemotherapy regimen and number of treatment cycles did not reveal significant variations, indicating an overall stability of sperm DNA methylation profiles over time. In the cross-sectional analysis, comparisons between patients and healthy controls identified significant differences for specific CpG sites of IGF2 and H19. Pre-treatment patients (T0) exhibited increased methylation at one CpG site of IGF2 and reduced methylation at multiple CpG sites of H19 compared with controls. At T2, the altered methylation pattern of H19 persisted, whereas the IGF2 difference was no longer significant. No significant differences were detected for PEG1/MEST and BCL2. Overall, the results suggest that chemotherapy does not induce biologically relevant alterations in the methylation status of the analyzed genes in sperm DNA from TGCT patients. These findings support previous evidence indicating limited epigenetic effects of cytotoxic treatments on imprinted genes. Ongoing analyses are investigating the SCAP gene, involved in embryonic development and lipid metabolism regulation, as a potential marker of chemotherapy-related epigenetic instability.
Assessment of Sperm DNA Methylation in Testicular Germ Cell Tumor Patients Before and After Chemotherapy
Francesca D’Angelo;Fani Konstantinidou;Liborio Stuppia;Barbara Barboni;
2026-01-01
Abstract
Testicular germ cell tumor (TGCT) patients are frequently exposed to cytotoxic therapies that may induce epigenetic alterations in sperm DNA, potentially affecting reproductive outcomes and embryonic development [1,2]. This study evaluated the methylation status of four genes involved in genomic imprinting and embryogenesis, PEG1/MEST, IGF2, H19, and BCL2, in sperm DNA from TGCT patients before and after chemotherapy [3]. Twenty-four TGCT patients were included in a longitudinal cohort providing semen samples before treatment (T0) and two years after therapy completion (T2). An additional cross-sectional cohort included 31 post-treatment patients and 10 fertile normozoospermic controls. Sperm DNA underwent bisulfite conversion followed by pyrosequencing analysis of CpG site within the promoter regions of the selected genes. In the longitudinal analysis, no statistically significant differences in methylation levels were observed for any of the analyzed genes. Similarly, stratification according to chemotherapy regimen and number of treatment cycles did not reveal significant variations, indicating an overall stability of sperm DNA methylation profiles over time. In the cross-sectional analysis, comparisons between patients and healthy controls identified significant differences for specific CpG sites of IGF2 and H19. Pre-treatment patients (T0) exhibited increased methylation at one CpG site of IGF2 and reduced methylation at multiple CpG sites of H19 compared with controls. At T2, the altered methylation pattern of H19 persisted, whereas the IGF2 difference was no longer significant. No significant differences were detected for PEG1/MEST and BCL2. Overall, the results suggest that chemotherapy does not induce biologically relevant alterations in the methylation status of the analyzed genes in sperm DNA from TGCT patients. These findings support previous evidence indicating limited epigenetic effects of cytotoxic treatments on imprinted genes. Ongoing analyses are investigating the SCAP gene, involved in embryonic development and lipid metabolism regulation, as a potential marker of chemotherapy-related epigenetic instability.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
