Alteration of RNA modification signature in human sperm correlates with sperm motility

Abstract

RNA modifications, which are introduced post-transcriptionally, have recently been assigned pivotal roles in the regulation of spermatogenesis and embryonic development. However, the RNA modification landscape in human sperm is poorly characterized, hampering our understanding about the potential role played by RNA modification in sperm. Through our recently developed high-throughput RNA modification detection platform based on liquid chromatography with tandem mass spectroscopy, we are the first to have characterized the RNA modification signature in human sperm. The RNA modification signature was generated on the basis of 49 samples from participants, including 13 healthy controls, 21 patients with asthenozoospermia (AZS) and 15 patients with teratozoospermia (TZS). In total, we identified 13 types of RNA modification marks on the total RNA in sperm, and 16 types of RNA modification marks on sperm RNA fragments of different sizes. The levels of these RNA modifications on the RNA of patients with AZS or TZS were altered, compared to controls, especially on sperm RNA fragments > 80 nt. A few types of RNA modifications, such as m1G, m5C, m2G and m1A, showed clear co-expression patterns as well as high linear correlations with clinical sperm motility. In conclusion, we characterized the RNA modification signature of human sperm and identified its correlation with sperm motility, providing promising candidates for use in clinical sperm quality assessment and new research insights for exploring the underlying pathological mechanisms in human male infertility syndromes.

Sperm RNA-mediated epigenetic inheritance in mammals: challenges and opportunities

“…Notably, a combined RNA modification signature (e.g. m 1 G, m 5 C, m 2 G and m 1 A) in sperm RNAs have been shown recently to be correlated with sperm motility, providing the potential for diagnostic value in IVF clinics (Guo et al 2022); other tissue-or blood-based RNA modification signatures are also implicated as biomarker under different disease conditions (Zhang et al , 2022.…”

METTL1 mediated tRNA m 7 G modification promotes leukaemogenesis of AML via tRNA regulated translational control

“…Interestingly, we found that the level of Gm, m 5 C, and Ψ modi cation on tRNA or mRNA were altered with m 7 G abundance declines in AML cells after METTL1 knockdown. We previous report that the abundances of different types of RNA modi cations exist extensive correlation in human sperm and mammalian tissue RNAs, such as m 5 C vs m 2 G and m 5 C vs m 7 G [32,54]. Although the underlying mechanisms have not been completely elucidated, the observation of non-METTL1-mediated RNA modi cations abundance alteration in this study provide new research insight of different RNA modi cation cooperation in regulating leukeamogenesis of AML.…”

“…Liquid chromatography-coupled mass spectrometry for the quantitative analysis of tRNA and mRNA modi cations Liquid chromatography-coupled mass spectrometry (LC-MS) was conducted as previously described [32]. Brie y, the RNAs concentration were measured, and 100 ng RNAs were digested in a 30 µl enzymolysis system including 3 µl 10 x buffer (250 mM Tirs-HCl, pH 8.0; 5 mM MgCl2 and 0.5 mg/mL BSA), 1 IU Benzonase (Sigma-Aldrich, catalog number: E8263, USA), 0.2 IU alkaline phosphatase (Sigma-Aldrich, catalog number: P5521, USA) and 0.05 IU phosphodiesterase I (USBiological, catalog number: P4072, USA) at 37 o C for 3 hours.…”

METTL1 mediated tRNA m7G modification promotes leukaemogenesis of AML via tRNA regulated translational control

“…Interestingly, we found that the level of Gm, m 5 C, I, and Ψ modification on tRNA or mRNA were altered with m 7 G abundance declines in AML cells after METTL1 knockdown. We previously reported that the abundances of different types of RNA modifications exist in extensive correlation in human sperm and mammalian tissue RNAs, such as m 5 C vs m 2 G and m 5 C vs m 7 G [ 38 , 62 ]. Although the underlying mechanisms have not been completely elucidated, the observation of non-METTL1-mediated RNA modifications abundance alteration in this study provides new research insights into different RNA modification cooperation in regulating leukaemogenesis of AML.…”

“…Liquid chromatography-coupled mass spectrometry (LC–MS/MS) was conducted as previously described [ 38 ]. Briefly, the RNAs concentration was measured, and 100 ng RNAs were digested in a 30 μl enzymolysis system including 3 μl 10 × buffer (250 mM Tirs-HCl, pH 8.0; 5 mM MgCl2 and 0.5 mg/ml BSA), 1 IU Benzonase (Sigma-Aldrich, catalog number: E8263, USA), 0.2 IU alkaline phosphatase (Sigma-Aldrich, catalog number: P5521, USA) and 0.05 IU phosphodiesterase I (USBiological, catalog number: P4072, USA) at 37 °C for 3 h. The enzymolysis samples then were centrifuged to obtain the mononucleotides using Nanosep® 3K spin filter (Pall Corporation, catalog number: 0D003C34, USA).…”