Ricerca Progetti nazionali, regionali, internazionaliNAZIONALIPRIN 2022Chemotherapy exposure of testicular cancer

Chemotherapy exposure of testicular cancer survivors: study of genetic and epigenetic alterations in spermatozoa

Testicular germ cell tumor (TGCT) is the most common cancer in men of reproductive age with a 5-year survival rate above 90% of patients. Since in over 95% of cases spermatogenesis will be recovered after 1-2 years, patients are prone to search for natural conception instead of undergoing to in vitro fertilization with pre-treatment cryopreserved spermatozoa. This implies that the identification of possible long-term effects of these treatments on fertility potential and reproductive safety are of paramount importance in the counselling of TGCT survivors. Up to date, only sperm DNA fragmentation and sperm aneuploidy rate have been evaluated as biomarkers of the genotoxic effect of chemotherapy in human. However, as anticipated by experiments carried out in rodents, these treatments are likely to induce a broader genotoxic effect such as de novo mutations and epigenetic alterations (e.g. DNA methylation and miRNAs) in spermatozoa. Data in the literature on de novo mutation rate is limited to 4 TGCT patients, whereas there are no data on DNA methylation and miRNA profiling in human. Given the paucity of data on the above issues, our collaborative study aims to get novel insights into the long-term effect of chemotherapy on sperm genome and epigenome. This objective will be achieved through the assessment of the following specific aims: i) the mutagenic effect of chemotherapy through the determination of the level of selfish mutations in the ejaculate via a novel droplet digital PCR (ddPCR) assay approach; ii) epigenetic modifications of chemotherapy in terms of A) sperm DNA methylation by methylome and target NGS analysis; B) sperm miRNA profiling by microfluidic miRNA cards and ddPCR assay. Semen analysis, clinical evaluation and sample collection for DNA extraction will be performed in the context of a longitudinal setting: i) for the mutation load analysis on 15 TGCT patients at baseline before chemotherapy (T0) and after 5 years (T5) from the end of therapy; ii) for the epigenetic studies on 15 TGCT at baseline before chemotherapy (T0), 2 years (T2) and 3 years (T3) after chemotherapy. Since this National Project requires a two-year involvement we will use previously collected baseline samples (T0). Three units (Rome, Florence and Chieti) with specific expertise in different molecular analyses will collaborate. The Rome and Florence units, two large andrology centers, will recruit patients. Sperm DNA from patients will be transferred to Chieti for methylation analysis. This project has remarkable potential implications in the clinical practice. The ultimate goal of our study is to provide personalized pre-conception counselling of TGCT survivors based on reliable biomarkers in order to help couples in their decision making i.e. whether to attempt conception with cryopreserved versus post-chemotherapy spermatozoa and what would be the most appropriate post-therapy time-interval for natural conception.

 

Data di avvio 28 Settembre 2023

Data di completamento 28 Settembre 2025

Total cost  € 88461,00

 

Progetto 2022EHN49M finanziato all’interno del Bando PRIN 2022 di cui al Decreto Direttoriale n. 104 del 02/02/2022 nell’ambito del Piano Nazionale di Ripresa e Resilienza, Missione 4 – Componente 2. Dalla Ricerca all’Impresa - Investimento 1.1 Fondo per il Programma Nazionale della Ricerca (PNR) e Progetti di Ricerca di Rilevante Interesse Nazionale (PRIN), finanziato dall’Unione europea – NextGenerationEU – CUP B53D23008040006