Identification of biomarkers for non-obstructive azoospermia based on microRNA and bioinformatics screening
Received date: 2025-07-02
Revised date: 2025-10-24
Online published: 2025-10-30
Supported by
Guangdong Basic and Applied Basic Research Foundation(2025A1515012629);Guangdong Basic and Applied Basic Research Foundation(2023A1515220129);Guangdong Basic and Applied Basic Research Foundation(2019A1515010249);Scientific Research Project of Guangdong Province Bureau of Traditional Chinese Medicine(20241387);Open Research Funds from the Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University(202301-306);Open Research Funds from the Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University(202301-104);Plan on Enhancing Innovation Capacity in Guangzhou Medical University(240603131131);Plan on Enhancing Innovation Capacity in Guangzhou Medical University(240603131129);Innovation training Program for university students in Guangdong province(S202410570053)
MicroRNAs (miRNAs) play an important role in the occurrence of non-obstructive azoospermia (NOA). Nevertheless, there is still a lack of research on the molecular mechanisms by which miRNAs regulate target genes to mediate NOA at present. In this study, we obtained NOA-related miRNA datasets from the GEO database and applied differential expression matrices combined with weighted correlation network analysis (WGCNA) and LASSO regression to identify four key miRNAs. Based on the miRDB database, the target genes of these miRNAs were predicted and intersected with the differentially expressed genes (DEGs) in the NOA transcriptome datasets. This intersection resulted in the identification of 18 DEGs. The spermatogenesis score model revealed a significant positive correlation between the overall expression level of these 18 DEGs and spermatogenesis scores, suggesting their potential involvement in NOA development. These 18 DEGs were subsequently incorporated into machine learning, leading to the identification of four hub genes with high diagnostic value: MGARP, FER1L5, SNX2, and PAPOLB. In the NOA mouse model, MGARP and SNX2 were upregulated, whereas FER1L5 and PAPOLB were downregulated, consistent with the expression trends observed in the NOA datasets. These findings indicate that MGARP, FER1L5, SNX2, and PAPOLB may serve as novel biomarkers for NOA, providing a theoretical and experimental foundations for elucidating its mechanisms and improving clinical diagnosis.
Zhihong Li, Miaoqi Chen, Xiaojun Yuan, Huajun Huang, Wanting Huang, Piaoyan Zhou, Chen Zeng, Xunuo Feng, Luoyao Yang, Shuqiang Huang, Cuiyu Tan, Cairong Chen, Qiuxia Yan . Identification of biomarkers for non-obstructive azoospermia based on microRNA and bioinformatics screening[J]. Hereditas(Beijing), 2026 , 48(3) : 301 -312 . DOI: 10.16288/j.yczz.25-089
| [1] | Chen YD, Liu XX, Zhang L, Zhu FY, Yan LY, Tang WH, Zhang Z, Liu Q, Jiang H, Qiao J. Deciphering the molecular characteristics of human Idiopathic nonobstructive azoospermia from the perspective of germ cells. Adv Sci (Weinh), 2023, 10(17): e2206852. |
| [2] | Zhang XY, Zhu TY, Zhang QR, Guo XJ, Wang C, Jin GF, Hu ZB. Progress in the genetic studies of spermatogenesis abnormalities. Hereditas(Beijing), 2021, 43(5): 473-486. |
| 张星雨, 祝天喻, 张清荣, 郭雪江, 王铖, 靳光付, 胡志斌. 精子发生障碍的遗传学研究进展. 遗传, 2021, 43(5): 473-486. | |
| [3] | Piechka A, Sparanese S, Witherspoon L, Hach F, Flannigan R. Molecular mechanisms of cellular dysfunction in testes from men with non-obstructive azoospermia. Nat Rev Urol, 2024, 21(2): 67-90. |
| [4] | Ruvkun G. Molecular biology. Glimpses of a tiny RNA world. Science, 2001, 294(5543): 797-799. |
| [5] | Ran ML, Chen B, Yin J, Yang AQ, Jiang M. Advances in miRNA research related to testis development and spermatogenesis. Hereditas (Beijing), 2014, 36(7): 646-654. |
| 冉茂良, 陈斌, 尹杰, 杨岸奇, 蒋明. 睾丸发育和精子生成相关miRNA研究进展. 遗传, 2014, 36(7): 646-654. | |
| [6] | Shi ZY, Yu M, Guo TC, Sui Y, Tian ZY, Ni X, Chen XR, Jiang M, Jiang JY, Lu YP, Lin MN. MicroRNAs in spermatogenesis dysfunction and male infertility: clinical phenotypes, mechanisms and potential diagnostic biomarkers. Front Endocrinol (Lausanne), 2024, 15: 1293368. |
| [7] | Sabetian S, Zarei M, Jahromi BN, Morowvat MH, Tabei SMB, Cava C. Exploring the dysregulated mRNAs- miRNAs-lncRNAs interactions associated to idiopathic non-obstructive azoospermia. J Biomol Struct Dyn, 2022, 40(13): 5956-5964. |
| [8] | Yu M, Mu HL, Niu ZW, Chu ZL, Zhu HJ, Hua JL. MiR-34c enhances mouse spermatogonial stem cells differentiation by targeting Nanos2. J Cell Biochem, 2014, 115(2): 232-242. |
| [9] | Wang L, Sun J, Han J, Ma ZW, Pan ML, Du ZJ. MiR-181a promotes spermatogenesis by targeting the S6K1 pathway. Int J Stem Cells, 2021, 14(3): 341-350. |
| [10] | Lian J, Zhang XS, Tian H, Liang N, Wang Y, Liang CZ, Li X, Sun F. Altered microRNA expression in patients with non-obstructive azoospermia. Reprod Biol Endocrinol, 2009, 7: 13. |
| [11] | Lin WW, Lamb DJ, Lipshultz LI, Kim ED. Demonstration of testicular apoptosis in human male infertility states using a DNA laddering technique. Int Urol Nephrol, 1999, 31(3): 361-370. |
| [12] | Finocchi F, Pelloni M, Balercia G, Pallotti F, Radicioni AF, Lenzi A, Lombardo F, Paoli D. Seminal plasma miRNAs in Klinefelter syndrome and in obstructive and non- obstructive azoospermia. Mol Biol Rep, 2020, 47(6): 4373-4382. |
| [13] | Tao Y. Endocrine aberrations of human nonobstructive azoospermia. Asian J Androl, 2022, 24(3): 274-286. |
| [14] | Zhang W, Zhang YN, Zhao MJ, Ding N, Yan L, Chen J, Gao LN, Zhang GZ, Sun XZ, Gu YQ, Liu ML. MicroRNA expression profiles in the seminal plasma of nonobstructive azoospermia patients with different histopathologic patterns. Fertil Steril, 2021, 115(5): 1197-1211. |
| [15] | Chen YH, Wang XW. MiRDB: an online database for prediction of functional microRNA targets. Nucleic Acids Res, 2020, 48(D1): D127-D131. |
| [16] | Omolaoye TS, Hachim MY, du Plessis SS. Using publicly available transcriptomic data to identify mechanistic and diagnostic biomarkers in azoospermia and overall male infertility. Sci Rep, 2022, 12(1): 2584. |
| [17] | Omolaoye TS, Omolaoye VA, Kandasamy RK, Hachim MY, Du Plessis SS. Omics and male Infertility: highlighting the application of transcriptomic data. Life (Basel), 2022, 12(2): 280. |
| [18] | Song WY, Meng H, Wang XG, Jin HX, Yao GD, Shi SL, Wu L, Zhang XY, Sun YP. Reduced microRNA-188-3p expression contributes to apoptosis of spermatogenic cells in patients with azoospermia. Cell Prolif, 2017, 50(1): e12297. |
| [19] | Yang C, Yao CC, Tian RH, Zhu ZJ, Zhao LY, Li P, Chen HX, Huang YH, Zhi EL, Gong YH, Xue YJ, Wang H, Yuan QQ, He ZP, Li Z. miR-202-3p regulates Sertoli cell proliferation, synthesis function, and apoptosis by targeting LRP6 and Cyclin D1 of Wnt/β-catenin signaling. Mol Ther Nucleic Acids, 2019, 14: 1-19. |
| [20] | Zhao LY, Yao CC, Xing XY, Jing T, Li P, Zhu ZJ, Yang C, Zhai J, Tian RH, Chen HX, Luo JQ, Liu NC, Deng ZW, Lin XH, Li N, Fang J, Sun J, Wang CC, Zhou Z, Li Z. Single-cell analysis of developing and azoospermia human testicles reveals central role of Sertoli cells. Nat Commun, 2020, 11(1): 5683. |
| [21] | Meinhardt A, Wilhelm B, Seitz J. Expression of mitochondrial marker proteins during spermatogenesis. Hum Reprod Update, 1999, 5(2): 108-119. |
| [22] | Qi SL, Wang YF, Zhou MX, Ge YX, Yan YB, Wang J, Zhang SSM, Zhang SP. A mitochondria-localized glutamic acid-rich protein (MGARP/OSAP) is highly expressed in retina that exhibits a large area of intrinsic disorder. Mol Biol Rep, 2011, 38(5): 2869-2877. |
| [23] | Ran LX, Gao ZX, Chen Q, Cui FM, Liu XL, Xue BX. Identification and validation of diagnostic signature genes in non-obstructive azoospermia by machine learning. Aging (Albany NY), 2023, 15(10): 4465-4480. |
| [24] | Zhong Q, Lazar CS, Tronchère H, Sato T, Meerloo T, Yeo M, Songyang Z, Emr SD, Gill GN. Endosomal localization and function of sorting nexin 1. Proc Natl Acad Sci USA, 2002, 99(10): 6767-6772. |
| [25] | Abdul-Ghani M, Hartman KL, Ngsee JK. Abstrakt interacts with and regulates the expression of sorting nexin-2. J Cell Physiol, 2005, 204(1): 210-218. |
| [26] | Kashiwabara SI, Tsuruta S, Okada K, Yamaoka Y, Baba T. Adenylation by testis-specific cytoplasmic poly(A) polymerase, PAPOLB/TPAP, is essential for spermatogenesis. J Reprod Dev, 2016, 62(6): 607-614. |
| [27] | Kashiwabara SI, Tsuruta S, Yamaoka Y, Oyama K, Iwazaki C, Baba T. PAPOLB/TPAP regulates spermiogenesis independently of chromatoid body-associated factors. J Reprod Dev, 2018, 64(1): 25-31. |
| [28] | Li JY, Ying YY, Qian YL, Chen JP, Huang Y, Liu J, Lv PP, Liu YF, Hu XL, Schilit SLP, Sheng JZ, Huang HF, Zhang D. BNC1 promotes spermatogenesis by regulating transcription of Ybx2 and Papolb via direct binding to their promotor elements. Reprod Sci, 2021, 28(3): 785-793. |
| [29] | Zohni K, Zhang X, Tan SL, Chan P, Nagano MC. The efficiency of male fertility restoration is dependent on the recovery kinetics of spermatogonial stem cells after cytotoxic treatment with busulfan in mice. Hum Reprod, 2012, 27(1): 44-53. |
| [30] | Hong YG, Yuan QC, Wang Y, Wang DQ, Guan XJ, Chen CD. Analysis of toxicity and mechanisms of busulfan in non-obstructive azoospermia: a genomic and toxicological approach integrating molecular docking, single-cell sequencing, and experimentation in vivo. Ecotoxicol Environ Saf, 2025, 291: 117878. |
| [31] | Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 2004, 116(2): 281-297. |
| [32] | Xia MM, Shen XY, Niu CM, Xia J, Sun HY, Zheng Y. MicroRNA regulates Sertoli cell proliferation and adhesion. Hereditas(Beijing), 2018, 40(9): 724-732. |
| 夏蒙蒙, 申雪沂, 牛长敏, 夏静, 孙红亚, 郑英. MicroRNA参与调控睾丸支持细胞的增殖与粘附功能. 遗传, 2018, 40(9): 724-732. | |
| [33] | Ehses J, Schlegel M, Schröger L, Schieweck R, Derdak S, Bilban M, Bauer K, Harner M, Kiebler MA. The dsRBP Staufen2 governs RNP assembly of neuronal Argonaute proteins. Nucleic Acids Res, 2022, 50(12): 7034-7047. |
| [34] | Yoda M, Kawamata T, Paroo Z, Ye XC, Iwasaki S, Liu QH, Tomari Y. ATP-dependent human RISC assembly pathways. Nat Struct Mol Biol, 2010, 17(1): 17-23. |
| [35] | Mills IG, Praefcke GJK, Vallis Y, Peter BJ, Olesen LE, Gallop JL, Butler PJG, Evans PR, McMahon HT. EpsinR: an AP1/clathrin interacting protein involved in vesicle trafficking. J Cell Biol, 2003, 160(2): 213-222. |
| [36] | Wu GF, Rouvière JO, Schmid M, Jensen TH. RNA 3′end tailing safeguards cells against products of pervasive transcription termination. Nat Commun, 2024, 15(1): 10446. |
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