Identification of a Methylation Pattern in the SNRPN Gene Promoter and its Association with Semen Abnormality Among Iraqi Males
Keywords:Methylation, SNRPN gene, semen abnormality, male infertility
Infertility considered as a multifactorial condition; the small nuclear ribonucleoprotein polypeptide N (SNRPN) gene is an imprinted gene. However, abnormal imprinting of this gene due to the methylation may result in abnormal function or silencing of the gene. Main aim of this study is to investigate the methylation present at the promoter of (SNRPN) gene and its role as a risk factor for male infertility. Sixty- three infertile males with age mean (32.28 ± 6.88 years) and 13 fertile males as a control age mean (34.07 ± 6.52 years) were investigated. Whole genomic DNA was extracted, DNA integrity was checked using β-globin gene as an internal control. The targeted region was amplified by polymerase chain reaction (PCR) technique. In addition, the SNRPN gene's promoter methylation was qualitatively detected using Real time polymerase chain reaction (qPCR) utilizing two sets of primers: methylated and un-methylated. Results reveled that all of the 63 infertile males were experiencing decrease in sperm concentration 9.42 ± 8.70 million/ml, reduced progressive motility 2.89 ± 5.45% as well as strange sperm morphology 27.06 ± 16.50%, while the values in the control group are normal. The results of the current investigation showed that the promoter of SNRPN was hypermethylated in some samples 22.7%, somewhat methylated in others 20.4%, and unmethylated in other samples 56.8% from infertile samples, while none of the 13 control samples had any methylation. These findings suggest that SNRPN gene may be associated with the negative changes in semen parameters, which could lead to male infertility.
D. Santi. S. De Vincentis, E. Magnani,. and G. Spaggiari, "Impairment of Sperm DNA Methylation in Male Infertility: A Meta-Analytic Study''. Journal of Andrology .5 (4): 695-703. 2017.
N. Garrido, and I. Hervás "Personalized medicine in infertile men. Urol". Clin. North Am., 47, 245-255. 2020.
C. Krausz,; A.R. Escamilla,; and C. Chianese, "Genetics of male infertility: from research to clinic" Reproduction. 150 (5):R159- 174, 2015.
H. Dong,; Y. Wang,; Z. Zou,; L. Chen,.; C. Shen,; S. Xu,; J. Zhang,; F. Zhao,.; S. Ge,; Q. Gao,; H. Hu,; M. Song, and W. Wang," Abnormal Methylation of Imprinted Genes and Cigarette Smoking: Assessment of Their Association with the Risk of Male Infertility" .Reproductive Sciences, 2016.
O. Okutman, M.B. Rhouma, M. Benkhalifa et al. "Genetic evaluation of patients with non-syndromic male infertility" J. Assist. Reprod. Genet., 35, 1939-1951, 2018. doi: doi: 10.1007/s10815-018-1301-7.
J. Stomper, J. C. Rotondo, G. Greve, and M. Lübbert, "Hypomethylating agents (HMA) for the treatment of acute myeloid leukemia and myelodysplastic syndromes: mechanisms of resistance and novel HMA-based therapies" Leukemia, 2021. doi: 10.1038/s41375-021-01218-0
S.S. Hammoud, J. Purwar, C. Pflueger, B.R. Cairns, and D.T. Carrell, "Alterations in sperm DNA methylation patterns at imprinted loci in two classes of infertility" Fertility and sterility .94 (5): 1728-1733, 2010.
A. Botezatu,; S. Razvan,; S. Demetra,; V. I. Iulia, and A. Gabriela, "Methylation Pattern of Methylene Tetrahydrofolate Reductase and Small Nuclear Ribonucleoprotein Polypeptide N Promoters in Oligoasthenospermia: A Case-Control Study: Reproductive BioMedicine Online. 28 (2): 225-31, 2014.
C. Nevin, and M. Carroll, "Sperm DNA methylation, infertility and transgenerational epigenetics" Journal of Human Genetics and Clinical Embryology.1, (4): 9-10, 2015.
World Health Organization "Examination and Processing of Human Semen" World Health Edition, (10): 286, 2010.
M. I. Mezaal "Molecular Detection and genotyping of human Papilloma virus and cervical cancer prediction among women with abnormal Pap smears in Baghdad" PhD. Dissertation. University of Baghdad, 2012.
CA. Eads, K. D. Danenberg, K. Kawakami, L. B. Saltz, C. Blake, D. Shibata, P.V, Danenberg, , P.W. Laird, "MethyLight: a high-throughput assay to measure DNA methylation" Nucleic Acids Res 28(8):E32, 2000.
Peng, H., Zhao, P., Liu, J., Zhang, J., Zhang, J., Wang, Y., et al. (2018). Novel epigenomic biomarkers of male infertility identified by methylation patterns of CpG sites within imprinting control regions of H19 and SNRPN genes. OMICS J. Integr. Biol. 22, 354-364. doi: 10.1089/omi.2018.0019
S. Gunes, M. A. Arslan, G. N. T. Hekim, and R. Asci, "The role of epigenetic in idiopathic male infertility" Journal of Assisted Reproduction and Genetics . 33 (5):553-569, 2016.
H. Lou, F. Le, M. Hu, X. Yang, L. Li, L. Wang, et al. "Aberrant DNA methylation of IGF2-H19 locus in human fetus and in spermatozoa from assisted reproductive technologies. Reprod. Sci. 26, 997-1004. doi: 10.1177/1933719118802052
J. C. Rotondo, C. Lanzillotti, C. Mazziotta, M. Tognon, & F. Martini, "Epigenetics of male infertility: The role of DNA methylation" Frontiers in Cell and Developmental Biology, 1822., 2021.
M. Lorgen-Ritchie, A.D. Murray, A. C. Ferguson-Smith, M. Richards, G.W. Horgan, L. H. Phillips, & P. Haggarty, "Imprinting methylation in SNRPN and MEST1 in adult blood predicts cognitive ability" Plos one, 14(2), e0211799, 2019.
A. Minor, C. Victor, and M. Sai, "Aberrant DNA methylation at imprinted genes in testicular sperm retrieved from men with obstructive azoospermia and post vasectomy reversal" Reproduction. 141(6):749-757, 2011.
How to Cite
Copyright (c) 2022 Mays H. Ali
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Articles accepted for publication in Al-Mustansiriyah Journal of Science (MJS) are protected under the Creative Commons Attribution 4.0 International License (CC-BY-NC). Authors of accepted articles are requested to sign a copyright release form prior to their article being published. All authors must agree to the submission, sign copyright release forms, and agree to be included in any correspondence between MJS and the authors before submitting a work to MJS. For personal or educational use, permission is given without charge to print or create digital copies of all or portions of a MJS article. However, copies must not be produced or distributed for monetary gain. It is necessary to respect the copyright of any parts of this work that are not owned by MJS.