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Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia

  • Authors:
    • Ling He
    • Xingwu Wu
    • Rongye Wu
    • Ping Guo
    • Wenfeng He
    • Wanlei Sun
    • He Chen
  • View Affiliations / Copyright

    Affiliations: Department of Geratology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Assisted Reproductive Center, Jiangxi Maternal and Child Health Hospital, Nanchang, Jiangxi 330006, P.R. China, Department of Clinical Medicine, Jiangxi Health Vocational College, Nanchang, Jiangxi 330052, P.R. China, Obstetrics and Gynecology, Yichun Maternal and Child Health Hospital, Jiangxi 336000, P.R. China, Jiangxi Key Laboratory of Molecular Medicine, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Clinical Laboratory, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
    Copyright: © He et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 347
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    Published online on: March 23, 2022
       https://doi.org/10.3892/etm.2022.11275
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Abstract

Asthenozoospermia (AZS) is characterized by reduced sperm motility and its pathogenesis remains poorly understood. Piwi‑interacting RNAs (piRNAs) have been indicated to serve important roles in spermatogenesis. However, little is known about the correlation of piRNA expression with AZS. In the present study, small RNA sequencing (small RNA‑seq) was performed on sperm samples from AZS patients and fertile controls. Reverse transcription‑quantitative (RT‑q) PCR was used to validate the small RNA‑seq results. Bioinformatics analyses were performed to predict the functions of differentially expressed piRNAs (DEpiRNAs). Logistic regression models were constructed and receiver operating characteristic curve (ROC) analysis was used to evaluate their diagnostic performance. A total of 114 upregulated and 169 downregulated piRNAs were detected in AZS patients. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that the DEpiRNAs were mainly associated with transcription, signal transduction, cell differentiation, metal ion binding and focal adhesion. These results were verified by RT‑qPCR analysis of eight selected piRNAs. The PCR results were consistent with the sequencing results in patients with AZS compared with controls in the first cohort. The expression of piR‑hsa‑32694, piR‑hsa‑26591, piR‑hsa‑18725 and piR‑hsa‑18586 was significantly upregulated in patients with AZS. The diagnostic power of the four piRNAs was further analyzed using ROC analysis; piR‑hsa‑26591 exhibited an area under the ROC curve (AUC) of 0.913 (95% CI: 0.795‑0.994). Logistic regression modelling and subsequent ROC analysis indicated that the combination of the 4 piRNAs achieved good diagnostic efficacy (AUC: 0.935).
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1 

Vander Borght M and Wyns C: Fertility and infertility: Definition and epidemiology. Clin Biochem. 62:2–10. 2018.PubMed/NCBI View Article : Google Scholar

2 

Wu X, Pan Y, Fang Y, Zhang J, Xie M, Yang F, Yu T, Ma P, Li W and Shu Y: The biogenesis and functions of piRNAs in human diseases. Mol Ther Nucleic Acids. 21:108–20. 2020.PubMed/NCBI View Article : Google Scholar

3 

Ozata DM, Gainetdinov I, Zoch A, O'Carroll D and Zamore PD: PIWI-interacting RNAs: Small RNAs with big functions. Nat Rev Genet. 20:89–108. 2019.PubMed/NCBI View Article : Google Scholar

4 

Charlesworth AG, Nitschko V, Renaud MS and Claycomb JM: PIWI puts spermatogenesis in its place. Dev Cell. 57:149–151. 2022.PubMed/NCBI View Article : Google Scholar

5 

Heyn H, Ferreira HJ, Bassas L, Bonache S, Sayols S, Sandoval J, Esteller M and Larriba S: Epigenetic disruption of the PIWI pathway in human spermatogenic disorders. PLoS One. 7(e47892)2012.PubMed/NCBI View Article : Google Scholar

6 

Kamaliyan Z, Pouriamanesh S, Soosanabadi M, Gholami M and Mirfakhraie R: Investigation of piwi-interacting RNA pathway genes role in idiopathic non-obstructive azoospermia. Sci Rep. 8(142)2018.PubMed/NCBI View Article : Google Scholar

7 

Hong Y, Wang C, Fu Z, Liang H, Zhang S, Lu M, Sun W, Ye C, Zhang CY, Zen K, et al: Systematic characterization of seminal plasma piRNAs as molecular biomarkers for male infertility. Sci Rep. 6(24229)2016.PubMed/NCBI View Article : Google Scholar

8 

Lu JC, Huang YF and Lü N: WHO laboratory manual for the examination and processing of human semen: Its applicability to andrology laboratories in China. Zhonghua Nan Ke Xue. 16:867–871. 2010.PubMed/NCBI(In Chinese).

9 

None. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. J Androl. 17(442)1996.

10 

Satoru K, Shigeru O, Kiyoshi K, Toshifumi K, Hideo M and Rihachi I: Purification of human sperm by a discontinuous Percoll density gradient with an innercolumn. Biol Reprod. 35:1059–1063. 1986.PubMed/NCBI View Article : Google Scholar

11 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar

12 

Martin M: Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J. 17:10–12. 2011.

13 

Gordon A and Hannon GJ: Fastx-toolkit. FASTQ/A short-reads pre-processing tools, 2010. http://hannonlabcshl.edu/fastx_toolkit.

14 

Patel RK and Jain M: NGS QC toolkit: A toolkit for quality control of next generation sequencing data. PLoS One. 7(e30619)2012.PubMed/NCBI View Article : Google Scholar

15 

Fu Y, He W, Zhou C, Fu X, Wan Q, He L and Wei B: Bioinformatics analysis of circRNA expression and construction of ‘circRNA-miRNA-mRNA’ competing endogenous RNAs networks in bipolar disorder patients. Front Genet. 12(718976)2021.PubMed/NCBI View Article : Google Scholar

16 

Wang J, Zhang P, Lu Y, Li Y, Zheng Y, Kan Y, Chen R and He S: piRBase: A comprehensive database of piRNA sequences. Nucleic Acids Res. 47:D175–D180. 2019.PubMed/NCBI View Article : Google Scholar

17 

Griffiths-Jones S, Saini HK, van Dongen S and Enright AJ: miRBase: Tools for microRNA genomics. Nucleic Acids Res. 36:D154–D158. 2008.PubMed/NCBI View Article : Google Scholar

18 

Griffiths-Jones S, Grocock RJ, van Dongen S, Bateman A and Enright AJ: miRBase: microRNA sequences, targets and gene nomenclature. Nucleic Acids Res. 34 (Database Issue):D140–D144. 2006.PubMed/NCBI View Article : Google Scholar

19 

Griffiths-Jones S, Bateman A, Marshall M, Khanna A and Eddy SR: Rfam: An RNA family database. Nucleic Acids Res. 31:439–441. 2003.PubMed/NCBI View Article : Google Scholar

20 

Love MI, Huber W and Anders S: Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15(550)2014.PubMed/NCBI View Article : Google Scholar

21 

Tino P: Basic properties and information theory of Audic-Claverie statistic for analyzing cDNA arrays. BMC Bioinformatics. 10(310)2009.PubMed/NCBI View Article : Google Scholar

22 

John B, Enright AJ, Aravin A, Tuschl T, Sander C and Marks DS: Human MicroRNA targets. PLoS Biol. 2(e363)2004.PubMed/NCBI View Article : Google Scholar

23 

Enright AJ, John B, Gaul U, Tuschl T, Sander C and Marks DS: MicroRNA targets in Drosophila. Genome Biol. 5(R1)2003.PubMed/NCBI View Article : Google Scholar

24 

Alexa A and Rahnenfuhrer J: topGO: Enrichment analysis for gene ontology. R package version. 28:2013.

25 

Manfrevola F, Chioccarelli T, Cobellis G, Fasano S, Ferraro B, Sellitto C, Marella G, Pierantoni R and Chianese R: CircRNA role and circRNA-dependent network (ceRNET) in asthenozoospermia. Front Endocrinol (Lausanne). 11(395)2020.PubMed/NCBI View Article : Google Scholar

26 

Lu H, Xu D, Wang P, Sun W, Xue X, Hu Y, Xie C and Ma Y: RNA-sequencing and bioinformatics analysis of long noncoding RNAs and mRNAs in the asthenozoospermia. Biosci Rep. 40(BSR20194041)2020.PubMed/NCBI View Article : Google Scholar

27 

Heidary Z, Zaki-Dizaji M, Saliminejad K and Khorram Khorshid HR: MicroRNA profiling in spermatozoa of men with unexplained asthenozoospermia. Andrologia. 51(e13284)2019.PubMed/NCBI View Article : Google Scholar

28 

Larriba E and Del Mazo J: An integrative piRNA analysis of mouse gametes and zygotes reveals new potential origins and gene regulatory roles. Sci Rep. 8(12832)2018.PubMed/NCBI View Article : Google Scholar

29 

Dai P, Wang X and Liu MF: A dual role of the PIWI/piRNA machinery in regulating mRNAs during mouse spermiogenesis. Sci China Life Sci. 63:447–449. 2020.PubMed/NCBI View Article : Google Scholar

30 

Gou LT, Kang JY, Dai P, Wang X, Li F, Zhao S, Zhang M, Hua MM, Lu Y, Zhu Y, et al: Ubiquitination-deficient mutations in human piwi cause male infertility by impairing histone-to-protamine exchange during spermiogenesis. Cell. 169:1090–1104.e13. 2017.PubMed/NCBI View Article : Google Scholar

31 

Hasuwa H, Ishino K and Siomi H: Human PIWI (HIWI) is an azoospermia factor. Sci China Life Sci. 61:348–350. 2018.PubMed/NCBI View Article : Google Scholar

32 

Xiao X, Mruk DD, Wong CK and Cheng CY: Germ cell transport across the seminiferous epithelium during spermatogenesis. Physiology (Bethesda). 29:286–298. 2014.PubMed/NCBI View Article : Google Scholar

33 

Yang CC, Lin YS, Hsu CC, Tsai MH, Wu SC and Cheng WT: Seasonal effect on sperm messenger RNA profile of domestic swine (Sus Scrofa). Anim Reprod Sci. 119:76–84. 2010.PubMed/NCBI View Article : Google Scholar

34 

Percipalle P: New insights into co-transcriptional sorting of mRNA for cytoplasmic transport during development. Semin Cell Dev Biol. 32:55–62. 2014.PubMed/NCBI View Article : Google Scholar

35 

López-Lemus UA, Garza-Guajardo R, Barboza-Quintana O, Rodríguez-Hernandez A, García-Rivera A, Madrigal-Pérez VM, Guzmán-Esquivel J, García-Labastida LE, Soriano-Hernández AD, Martínez-Fierro ML, et al: Association between nonalcoholic fatty liver disease and severe male reproductive organ impairment (germinal epithelial loss): Study on a mouse model and on human patients. Am J Mens Health. 12:639–648. 2018.PubMed/NCBI View Article : Google Scholar

36 

Li Y, Liu L, Wang B, Xiong J, Li Q, Wang J and Chen D: Impairment of reproductive function in a male rat model of non-alcoholic fatty liver disease and beneficial effect of N-3 fatty acid supplementation. Toxicol Lett. 222:224–232. 2013.PubMed/NCBI View Article : Google Scholar

37 

Baeza R, Mazzeo I, Vílchez MC, Gallego V, Peñaranda DS, Pérez L and Asturiano JF: Relationship between sperm quality parameters and the fatty acid composition of the muscle, liver and testis of European eel. Comp Biochem Physiol A Mol Integr Physiol. 181:79–86. 2015.PubMed/NCBI View Article : Google Scholar

38 

Dougherty GW, Mizuno K, Nöthe-Menchen T, Ikawa Y, Boldt K, Ta-Shma A, Aprea I, Minegishi K, Pang YP, Pennekamp P, et al: CFAP45 deficiency causes situs abnormalities and asthenospermia by disrupting an axonemal adenine nucleotide homeostasis module. Nat Commun. 11(5520)2020.PubMed/NCBI View Article : Google Scholar

39 

Jodar M, Kalko S, Castillo J, Ballescà JL and Oliva R: Differential RNAs in the sperm cells of asthenozoospermic patients. Hum Reprod. 27:1431–1438. 2012.PubMed/NCBI View Article : Google Scholar

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Copy and paste a formatted citation
Spandidos Publications style
He L, Wu X, Wu R, Guo P, He W, Sun W and Chen H: Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia. Exp Ther Med 23: 347, 2022.
APA
He, L., Wu, X., Wu, R., Guo, P., He, W., Sun, W., & Chen, H. (2022). Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia. Experimental and Therapeutic Medicine, 23, 347. https://doi.org/10.3892/etm.2022.11275
MLA
He, L., Wu, X., Wu, R., Guo, P., He, W., Sun, W., Chen, H."Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia". Experimental and Therapeutic Medicine 23.5 (2022): 347.
Chicago
He, L., Wu, X., Wu, R., Guo, P., He, W., Sun, W., Chen, H."Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia". Experimental and Therapeutic Medicine 23, no. 5 (2022): 347. https://doi.org/10.3892/etm.2022.11275
Copy and paste a formatted citation
x
Spandidos Publications style
He L, Wu X, Wu R, Guo P, He W, Sun W and Chen H: Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia. Exp Ther Med 23: 347, 2022.
APA
He, L., Wu, X., Wu, R., Guo, P., He, W., Sun, W., & Chen, H. (2022). Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia. Experimental and Therapeutic Medicine, 23, 347. https://doi.org/10.3892/etm.2022.11275
MLA
He, L., Wu, X., Wu, R., Guo, P., He, W., Sun, W., Chen, H."Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia". Experimental and Therapeutic Medicine 23.5 (2022): 347.
Chicago
He, L., Wu, X., Wu, R., Guo, P., He, W., Sun, W., Chen, H."Seminal plasma piRNA array analysis and identification of possible biomarker piRNAs for the diagnosis of asthenozoospermia". Experimental and Therapeutic Medicine 23, no. 5 (2022): 347. https://doi.org/10.3892/etm.2022.11275
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