Overexpression of hsa-miR-30a-5p and nonobstructive azoospermia: A case-control study

Abstract

Background: Some previous human and animal studies have supported the idea that KDM3A down-regulation might be the main cause of male infertility, especially in nonobstructive azoospermia (NOA). The regulatory role of micro-RNAs (miRNA) has been investigated in the development of male infertility.


Objective: The expression level of hsa-miR-30a-5p in azoospermia was evaluated to reveal its possible association with the etiology of male infertility.


Materials and Methods: In this case-control study, 30 men with azoospermia (19 of whom had NOA) were selected as the case individuals, and 11 men with obstructive azoospermia (OA) were selected as control individuals. The best miRNA with the strongest ability to target the KDM3A gene was detected via comprehensive bioinformatics analysis. Reverse transcriptase quantitative polymerase chain reaction was used to assess the expression level of hsa-miR-30a-5p. After analyzing the data, the expression level of hsa-miR-30a-5p was compared between men with NOA and men with OA.


Results: The findings supported the idea that hsa-miR-30a-5p is the miRNA with the best ability to target the KDM3A transcript. The expression analysis of hsa-miR-30a-5p indicated a significant overexpression (p = 0.04) in men with NOA compared to in men with OA.


Conclusion: Hsa-miR-30a-5p was overexpressed in men with NOA compared to in control individuals. Hsa-miR-30a-5p could target the KDM3A transcript and may suppress its expression.


Key words: Hsa-miR-30a-5p, Male infertility, KDM3A, Azoospermia, miRNA.

References
[1] Fernández Pelegrina R, Kessler AG, Rawlins RG. Modern approaches to the treatment of human infertility through assisted reproduction. P R Health Sci J 1991; 10: 75–81.

[2] Irvine DS. Epidemiology and aetiology of male infertility. Hum Reprod 1998; 13 (Suppl.): 33–44.

[3] Aziz N. The importance of semen analysis in the context of azoospermia. Clinics 2013; 68 (Suppl.): 35–38.

[4] Jarow JP, Espeland MA, Lipshultz LI. Evaluation of the azoospermic patient. J Urol 1989; 142: 62–65.

[5] Javadirad SM, Hojati Z, Ghaedi K, Nasr-Esfahani MH. Expression ratio of histone demethylase KDM 3A to protamine-1 mRNA is predictive of successful testicular sperm extraction in men with obstructive and nonobstructive azoospermia. Andrology 2016; 4: 492–499.

[6] Okada Y, Scott G, Ray MK, Mishina Y, Zhang Y. Histone demethylase JHDM2A is critical for Tnp1 and Prm1 transcription and spermatogenesis. Nature 2007; 450: 119–123.

[7] Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 2010; 11: 597–610.

[8] Momeni A, Najafipour R, Hamta A, Jahani S, Moghbelinejad S. Expression and methylation pattern of hsa-miR-34 family in sperm samples of infertile men. Reprod Sci 2020; 27: 301–308.

[9] Wang Ch, Yang C, Chen X, Yao B, Yang Ch, Zhu Ch, et al. Altered profile of seminal plasma microRNAs in the molecular diagnosis of male infertility. Clin Chem 2011; 57: 1722–1731.

[10] Liu T, Huang Y, Liu J, Zhao Y, Jiang L, Huang Q, et al. MicroRNA-122 influences the development of sperm abnormalities from human induced pluripotent stem cells by regulating TNP2 expression. Stem Cells Dev 2013; 22: 1839–1850.

[11] Yan N, Lu Y, Sun H, Qiu W, Tao D, Liu Y, et al. Microarray profiling of microRNAs expressed in testis tissues of developing primates. J Assist Reprod Genet 2009; 26: 179–186.

[12] Kotaja N. MicroRNAs and spermatogenesis. Fertil Steril 2014; 101: 1552–1562.

[13] Walker WH. Regulation of mammalian spermatogenesis by miRNAs. Semin Cell Dev Biol 2021; S1084–9521(21)00118- X.

[14] Charan J, Biswas T. How to calculate sample size for different study designs in medical research? Indian J Psychol Med 2013; 35: 121–126.

[15] Norioun H, Motovali-Bashi M, Javadirad SM. Hsa-miR- 27a-3p overexpression in men with nonobstructive azoospermia: A case-control study. Int J Reprod BioMed 2020; 18: 961–968.

[16] Bonaparte E, Moretti M, Colpi GM, Nerva F, Contalbi G, Vaccalluzzo L, et al. ESX1 gene expression as a robust marker of residual spermatogenesis in azoospermic men. Hum Reprod 2010; 25: 1398–1403.

[17] Pansa A, Sirchia SM, Melis S, Giacchetta D, Castiglioni M, Colapietro P, et al. ESX1 mRNA expression in seminal fluid is an indicator of residual spermatogenesis in nonobstructive azoospermic men. Hum Reprod 2014; 29: 2620–2627.

[18] Lian J, Zhang X, Tian H, Liang N, Wang Y, Liang C, et al. Altered microRNA expression in patients with nonobstructive azoospermia. Reprod Biol Endocrinol 2009; 7: 13.

[19] Abu-Halima M, Hammadeh M, Schmitt J, Leidinger P, Keller A, Meese E, et al. Altered microRNA expression profiles of human spermatozoa in patients with different spermatogenic impairments. Fertil Steril 2013; 99: 1249– 1255.

[20] Abu-Halima M, Galata V, Backes C, Keller A, Hammadeh M, Meese E. MicroRNA signature in spermatozoa and seminal plasma of proven fertile men and in testicular tissue of men with obstructive azoospermia. Andrologia 2020; 52: e13503.

[21] Barad O, Meiri E, Avniel A, Aharonov R, Barzilai A, Bentwich I, et al. MicroRNA expression detected by oligonucleotide microarrays: System establishment and expression profiling in human tissues. Genome Res 2004; 14: 2486–2494.

[22] Yu Z, Raabe T, Hecht NB. MicroRNA Mirn122a reduces expression of the posttranscriptionally regulated germ cell transition protein 2 (Tnp2) messenger RNA (mRNA) by mRNA cleavage. Biol Reprod 2005; 73: 427–433.

[23] Lian J, Zhang X, Tian H, Liang N, Wang Y, Liang C, et al. Altered microRNA expression in patients with nonobstructive azoospermia. Reprod Biol Endocrinol 2009; 7: 1–10.