Identification of biomarkers of male infertility through the circRNA expression profiling of seminal plasma

Zhaode Liu , Xinrui Li , Xiaoyu Yang , Bohang Zhang , Dingdong Chen , Yan Yuan , Yiqiang Cui

Journal of Biomedical Research ›› 2025, Vol. 39 ›› Issue (4) : 367 -381.

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Journal of Biomedical Research ›› 2025, Vol. 39 ›› Issue (4) :367 -381. DOI: 10.7555/JBR.38.20240192
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Identification of biomarkers of male infertility through the circRNA expression profiling of seminal plasma
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Abstract

Circular RNAs (circRNAs) are key regulators of reproductive biology. However, limited information is available regarding circRNA expression profiles in seminal plasma samples from individuals with male infertility. The present study aimed to identify circRNAs associated with infertility in seminal plasma samples and to clarify their potential as biomarkers, as well as the possible molecular mechanisms underlying their functions. Next-generation RNA sequencing was conducted to analyze circRNA profiles in seminal plasma from healthy controls, oligoasthenospermia (OAZ) patients, and non-obstructive azoospermia (NOA) patients. Bioinformatics analysis revealed that 637 circRNAs were differentially expressed between OAZ and control subjects, and 272 circRNAs were differentially expressed between NOA and control subjects. The expression of key circRNAs (hsa-SAP130_0002, hsa-TRPC1_0001, hsa-FBRS_0001, hsa-ACACA_0025, hsa-UTRN_0042, and hsa-ZNF532_0023) was then validated by qPCR, and their diagnostic accuracy for infertility was confirmed through receiver operating characteristic curve analysis. Additionally, a possible circRNA-miRNA-mRNA regulatory network was constructed for these candidate biomarkers. Collectively, this study identifies a novel set of circRNAs with potential as diagnostic biomarkers for male infertility and provides molecular insights that may facilitate both diagnostic and therapeutic efforts.

Keywords

seminal plasma / circular RNA / male infertility / RNA-seq

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Zhaode Liu, Xinrui Li, Xiaoyu Yang, Bohang Zhang, Dingdong Chen, Yan Yuan, Yiqiang Cui. Identification of biomarkers of male infertility through the circRNA expression profiling of seminal plasma. Journal of Biomedical Research, 2025, 39(4): 367-381 DOI:10.7555/JBR.38.20240192

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Fundings

This work was funded by the National Key R&D Program of China (Grant No. 2022YFC2702800 to Y. C.), the National Natural Science Foundation of China (Grant No. 82122025 to Y. Y.), and the National Key R&D Program of China (Grant No. 2021YFC2700200 to Y. C.).

Acknowledgments

We would like to thank all the research subjects for providing valuable samples.

References

[1]

Inhorn MC, Patrizio P. Infertility around the globe: New thinking on gender, reproductive technologies and global movements in the 21st century[J]. Hum Reprod Update, 2015, 21(4): 411-426. doi: 10.1093/humupd/dmv016

[2]

Kimmins S, Anderson RA, Barratt CLR, et al. Frequency, morbidity and equity—the case for increased research on male fertility[J]. Nat Rev Urol, 2024, 21(2): 102-124. doi: 10.1038/s41585-023-00820-4

[3]

Eisenberg ML, Esteves SC, Lamb DJ, et al. Male infertility[J]. Nat Rev Dis Primers, 2023, 9(1): 49. doi: 10.1038/s41572-023-00459-w

[4]

Ma L, Chu H, Wang M, et al. Biological functions and potential implications of circular RNAs[J]. J Biomed Res, 2022, 37(2): 89-99. doi: 10.7555/JBR.36.20220095.

[5]

Wang F, Yang W, Ouyang S, et al. The vehicle determines the destination: The significance of seminal plasma factors for male fertility[J]. Int J Mol Sci, 2020, 21(22): 8499. doi: 10.3390/ijms21228499

[6]

Dong W, Li H, Qing X, et al. Identification and characterization of human testis derived circular RNAs and their existence in seminal plasma[J]. Sci Rep, 2016, 6: 39080. doi: 10.1038/srep39080

[7]

Cheng L, Jin H, Xiao T, et al. Human circBOULE RNAs as potential biomarkers for sperm quality and male infertility[J]. J Biomed Res, 2024, 38(5): 473-484. doi: 10.7555/JBR.37.20230296

[8]

Yue D, Yang R, Xiong C, et al. Functional prediction and profiling of exosomal circRNAs derived from seminal plasma for the diagnosis and treatment of oligoasthenospermia[J]. Exp Ther Med, 2022, 24(5): 649. doi: 10.3892/etm.2022.11586

[9]

Ji C, Wang Y, Wei X, et al. Potential of testis-derived circular RNAs in seminal plasma to predict the outcome of microdissection testicular sperm extraction in patients with idiopathic non-obstructive azoospermia[J]. Hum Reprod, 2021, 36(10): 2649-2660. doi: 10.1093/humrep/deab196

[10]

Huang S, Li H, Ding X, et al. Presence and characterization of cell-free seminal RNA in healthy individuals: Implications for noninvasive disease diagnosis and gene expression studies of the male reproductive system[J]. Clin Chem, 2009, 55(11): 1967-1976. doi: 10.1373/clinchem.2009.131128

[11]

Di L, Fu Y, Sun Y, et al. RNA sequencing by direct tagmentation of RNA/DNA hybrids[J]. Proc Natl Acad Sci U S A, 2020, 117(6): 2886-2893. doi: 10.1073/pnas.1919800117

[12]

Gao Y, Wang J, Zhao F. CIRI: An efficient and unbiased algorithm for de novo circular RNA identification[J]. Genome Biol, 2015, 16(1): 4. doi: 10.1186/s13059-014-0571-3

[13]

Memczak S, Jens M, Elefsinioti A, et al. Circular RNAs are a large class of animal RNAs with regulatory potency[J]. Nature, 2013, 495(7441): 333-338. doi: 10.1038/nature11928

[14]

Zhang X, Dong R, Zhang Y, et al. Diverse alternative back-splicing and alternative splicing landscape of circular RNAs[J]. Genome Res, 2016, 26(9): 1277-1287. doi: 10.1101/gr.202895.115

[15]

Zhang S, Wang C, Wang Y, et al. A novel protein encoded by circRsrc1 regulates mitochondrial ribosome assembly and translation during spermatogenesis[J]. BMC Biol, 2023, 21(1): 94. doi: 10.1186/s12915-023-01597-z

[16]

Liu C, Chen L. Circular RNAs: Characterization, cellular roles, and applications[J]. Cell, 2022, 185(12): 2016-2034. doi: 10.1016/j.cell.2022.04.021

[17]

Jeck WR, Sorrentino JA, Wang K, et al. Circular RNAs are abundant, conserved, and associated with ALU repeats[J]. RNA, 2013, 19(2): 141-157. doi: 10.1261/rna.035667.112

[18]

Zhang J, Hou L, Zuo Z, et al. Comprehensive profiling of circular RNAs with nanopore sequencing and CIRI-long[J]. Nat Biotechnol, 2021, 39(7): 836-845. doi: 10.1038/s41587-021-00842-6

[19]

Danan M, Schwartz S, Edelheit S, et al. Transcriptome-wide discovery of circular RNAs in Archaea[J]. Nucleic Acids Res, 2012, 40(7): 3131-3142. doi: 10.1093/nar/gkr1009

[20]

Bose R, Ain R. Regulation of transcription by circular RNAs[J]. Adv Exp Med Biol, 2018, 1087: 81-94. https://pubmed.ncbi.nlm.nih.gov/30259359/

[21]

Tang Z, Li X, Zhao J, et al. TRCirc: A resource for transcriptional regulation information of circRNAs[J]. Brief Bioinform, 2019, 20(6): 2327-2333. doi: 10.1093/bib/bby083

[22]

Paiardi C, Pasini ME, Gioria M, et al. Failure of acrosome formation and globozoospermia in the wobbler mouse, a Vps54 spontaneous recessive mutant[J]. Spermatogenesis, 2011, 1(1): 52-62. doi: 10.4161/spmg.1.1.14698

[23]

Tang EI, Cheng CY. MARK2 and MARK4 regulate sertoli cell BTB dynamics through microtubule and actin cytoskeletons[J]. Endocrinology, 2022, 163(11): bqac130. doi: 10.1210/endocr/bqac130

[24]

Bell EL, Nagamori I, Williams EO, et al. SirT1 is required in the male germ cell for differentiation and fecundity in mice[J]. Development, 2014, 141(18): 3495-3504. doi: 10.1242/dev.110627

[25]

Bose R, Sheng K, Moawad AR, et al. Ubiquitin ligase Huwe1 modulates spermatogenesis by regulating spermatogonial differentiation and entry into meiosis[J]. Sci Rep, 2017, 7(1): 17759. doi: 10.1038/s41598-017-17902-0

[26]

Dwyer JL, Richburg JH. Age-dependent alterations in spermatogenesis in itchy mice[J]. Spermatogenesis, 2012, 2(2): 104-116. doi: 10.4161/spmg.20596

[27]

Sinha N, Whelan EC, Tobias JW, et al. Roles of Stra8 and Tcerg1l in retinoic acid induced spermatogonial differentiation in mouse[J]. Biol Reprod, 2021, 105(2): 503-518. doi: 10.1093/biolre/ioab093

[28]

Li J, Wang X, Shi L, et al. A mammalian conserved circular RNA circLARP1B regulates hepatocellular carcinoma metastasis and lipid metabolism[J]. Adv Sci (Weinh), 2024, 11(2): e2305902. doi: 10.1002/advs.202305902

[29]

Heumüller AW, Jones AN, Mourão A, et al. Locus-conserved circular RNA cZNF292 controls endothelial cell flow responses[J]. Circ Res, 2022, 130(1): 67-79. doi: 10.1161/CIRCRESAHA.121.320029

[30]

Hansen EB, Fredsøe J, Okholm TLH, et al. The transcriptional landscape and biomarker potential of circular RNAs in prostate cancer[J]. Genome Med, 2022, 14(1): 8. doi: 10.1186/s13073-021-01009-3

[31]

Chen HC, Chin YF, Lundy DJ, et al. Utrophin compensates dystrophin loss during mouse spermatogenesis[J]. Sci Rep, 2017, 7(1): 7372. doi: 10.1038/s41598-017-05993-8

[32]

Sun Q, Liang R, Li M, et al. Circ_UTRN ameliorates caerulein-induced acute pancreatitis in vitro via reducing inflammation and promoting apoptosis through miR-320-3p/PTK2 axis[J]. J Pharm Pharmacol, 2022, 74(6): 861-868. doi: 10.1093/jpp/rgab161

[33]

Hansen TB, Jensen TI, Clausen BH, et al. Natural RNA circles function as efficient microRNA sponges[J]. Nature, 2013, 495(7441): 384-388. doi: 10.1038/nature11993

[34]

de Gendt K, Swinnen JV, Saunders PTK, et al. A Sertoli cell-selective knockout of the androgen receptor causes spermatogenic arrest in meiosis[J]. Proc Natl Acad Sci U S A, 2004, 101(5): 1327-1332. doi: 10.1073/pnas.0308114100

[35]

Mozdarani H, Ghoraeian P, Mozdarani S, et al. High frequency of de novo DAZ microdeletion in sperm nuclei of subfertile men: Possible involvement of genome instability in idiopathic male infertility[J]. Hum Fertil (Camb), 2018, 21(2): 137-145. doi: 10.1080/14647273.2017.1322718

[36]

Dowdle JA, Mehta M, Kass EM, et al. Mouse BAZ1A (ACF1) is dispensable for double-strand break repair but is essential for averting improper gene expression during spermatogenesis[J]. PLoS Genet, 2013, 9(11): e1003945. doi: 10.1371/journal.pgen.1003945

[37]

Chen K, Wei B, Hao S, et al. The PI3K/AKT signaling pathway: How does it regulate development of Sertoli cells and spermatogenic cells?[J]. Histol Histopathol, 2022, 37(7): 621-636. https://pubmed.ncbi.nlm.nih.gov/35388905/

[38]

Xue R, Lin W, Sun J, et al. The role of Wnt signaling in male reproductive physiology and pathology[J]. Mol Hum Reprod, 2021, 27(1): gaaa085. doi: 10.1093/molehr/gaaa085

[39]

Zhu F, Luo Y, Bo H, et al. Trace the profile and function of circular RNAs in Sertoli cell only syndrome[J]. Genomics, 2021, 113(4): 1845-1854. doi: 10.1016/j.ygeno.2021.04.022

[40]

Bo H, Liu Z, Tang R, et al. Testicular biopsies microarray analysis reveals circRNAs are involved in the pathogenesis of non-obstructive azoospermia[J]. Aging, 2020, 12(3): 2610-2625. doi: 10.18632/aging.102765

[41]

Kohansal M, Alghanimi YK, Banoon SR, et al. CircRNA-associated ceRNA regulatory networks as emerging mechanisms governing the development and biophysiopathology of epilepsy[J]. CNS Neurosci Ther, 2024, 30(4): e14735. doi: 10.1111/cns.14735

[42]

Zeng Y, Zou Y, Gao G, et al. The biogenesis, function and clinical significance of circular RNAs in breast cancer[J]. Cancer Biol Med, 2021, 19(1): 14-29. doi: 10.20892/j.issn.2095-3941.2020.0485

[43]

Tan C, Xie G, Wu S, et al. Simultaneous detection of breast cancer biomarkers circROBO1 and BRCA1 based on a CRISPR-Cas13a/Cas12a system[J]. Biosens Bioelectron, 2024, 258: 116373. doi: 10.1016/j.bios.2024.116373

[44]

Sun Z, Yang C, Huang L, et al. circRNADisease v2.0: An updated resource for high-quality experimentally supported circRNA-disease associations[J]. Nucleic Acids Res, 2024, 52(D1): D1193-D1200. doi: 10.1093/nar/gkad949

[45]

Ou N, Wang Y, Xu S, et al. Primate-specific DAZ regulates translation of cell proliferation-related mRNAs and is essential for maintenance of spermatogonia[J]. Adv Sci (Weinh), 2024, 11(29): e2400692. doi: 10.1002/advs.202400692

[46]

Zhou F, Yuan Q, Zhang W, et al. MiR-663a stimulates proliferation and suppresses early apoptosis of human spermatogonial stem cells by targeting NFIX and regulating cell cycle[J]. Mol Ther Nucleic Acids, 2018, 12: 319-336. doi: 10.1016/j.omtn.2018.05.015

[47]

Wang Y, Li X, Gong X, et al. MicroRNA-322 regulates self-renewal of mouse spermatogonial stem cells through Rassf8[J]. Int J Biol Sci, 2019, 15(4): 857-869. doi: 10.7150/ijbs.30611

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