Evaluantion of small non-coding RNA levels in spermatozoa from patients with negative outcomes in assisted reporductive technologies

GROSSO, J; CLAUS, C; CALVO, K; BRIGNARDELLO, C; MORENTE, C; SPINELLI, S.V.

Mar del Plata

Congreso; Reunión SAI-SAIC-SAFIS 2022; 2022

Fertility problems affect 15% of couples worldwide and the causes can be attributed equally to the male or female partner. Since conventional semen analysis provides a limited prediction of male fertility, in our laboratory we evaluated the potential use of sRNAs as markers of semen quality in assisted reproduction treatments (ART). Based on previous research, we focused on tRNA-derived fragments produced by the ribonuclease angiogenine, called tiRNAs. These molecules are key regulators of physiological and pathological processes with a great impact on reproductive function. In this work, we analyze sperm cells (SPZ) to assess their quality and possible associations with seminal tiRNAs and ART results. We conducted a prospective study with couples who underwent ART treatments with donated oocytes at the PROAR Medical Center between 2018-2019. Samples were collected from normozoospermic men and different transcripts were quantified by RT-qPCR (N=17, 10 ART+ and 7 ART-). Data was statistically evaluated using the Mann Whitney test.As endogenous controls we employed U6 for sRNA and PPIA for mRNA. Even though reactions were prepared using the same amount of total RNA, levels of both transcripts showed strong dispersion, with differences in Ct of more than 10 cycles for U6. These results indicate that RNAs stability in mature SPZ is highly variable between samples. More interestingly, levels of U6 in SPZ tend to be lower in patients with failed ART cycles, showing a discrete group with the lowest levels that could potentially account to patients with unexplained infertility. Levels of tiRNAGlu, tiRNAGly and tiRNALys showed no differences between groups, but their absolute levels correlate with U6 expression only in samples form successful ART (. We also tried to measure the angiogenin transcript but it was not detected in the SPZ,suggesting that sperm tiRNAs may be incorporated from seminla plasma . The sperm paturation marker SLFNL1, GAPDHS and ZBPB1 were also quantified > No significant differences were observed between the groups but GAPDHS and ZBPB1showed higher detection in samples from patients whose couples reach pregnancy. These results provides preliminary data showing the potential role of RNA stability and tiRNAs in male infertility.