title: The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia.
abstract: Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia. Initially, through analyzing CRISP2 expression and methylation on CRISP2 promoter activity in sperm, we observed a decreased expression of CRISP2 protein rather than its mRNA in the ejaculated spermatozoa from asthenozoospermic patients and no methylation in the CRISP2 promoter, suggesting CRISP2 expression may be regulated in the sperm at the posttranscriptional level. Subsequently, we found that microRNA 27b (miR-27b), predicted as a candidate regulator of CRISP2 using bioinformatics, was highly expressed in the ejaculated spermatozoa from asthenozoospermic patients. Luciferase reporter assay and transfection experiments disclosed that this microRNA could target CRISP2 by specifically binding its 3' untranslated region, suppressing CRISP2 expression. Extended clinical observation further confirmed a highly expressed miR-27b and its obviously negative correlation with CRISP2 protein expression in ejaculated spermatozoa samples from asthenozoospermic patients. Finally, we conducted a retrospective follow-up study to support that either high miR-27b expression or low CRISP2 protein expression was significantly associated with low sperm progressive motility, abnormal morphology, and infertility. Thus, this study provides the first preliminary insight into the mechanism leading to the reduced CRISP2 expression in asthenozoospermia, offering a potential therapeutic target for treating male infertility or for male contraception.
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Jun-Hao
authors.initials: JH
authors.lastname: Zhou
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Qi-Zhao
authors.initials: QZ
authors.lastname: Zhou
authors.affiliation: Medical Laboratory Centre, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China Cancer Research Institute, Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Lyu
authors.affiliation: Cancer Research Institute, Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhu
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Zi-Jian
authors.initials: ZJ
authors.lastname: Chen
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Ming-Kun
authors.initials: MK
authors.lastname: Chen
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Xia
authors.affiliation: Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Qi
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China Cancer Research Institute, Southern Medical University, Guangzhou, Guangdong, China xinli268@gmail.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong, China cundongliu@163.com.
authors.forename: Cun-Dong
authors.initials: CD
authors.lastname: Liu
mesh_terms: D000328:Adult; D053627:Asthenozoospermia / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D016022:Case-Control Studies; D015815:Cell Adhesion Molecules; D002478:Cells, Cultured; D019175:DNA Methylation; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012189:Retrospective Studies; D013094:Spermatozoa / Q000378:metabolism*; D055815:Young Adult
chemical_list: C062621:CRISP2 protein, human; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; C531388:MIRN27 microRNA, human; D035683:MicroRNAs
keywords: asthenozoospermia; cysteine-rich secretory protein 2 (CRISP2); male infertility; miR-27b