{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T12:20:55Z","timestamp":1754482855805,"version":"3.40.5"},"reference-count":32,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2023,6,19]],"date-time":"2023-06-19T00:00:00Z","timestamp":1687132800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2018YFC1003603"],"award-info":[{"award-number":["2018YFC1003603"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Andrology"],"published-print":{"date-parts":[[2024,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>The ubiquitin ligase HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 is essential for the establishment and maintenance of spermatogonia. However, the role of HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 in regulating germ cell differentiation remains unclear, and clinical evidence linking HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 to male infertility pathogenesis is lacking.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>This study aims to investigate the role of <jats:italic>HUWE1<\/jats:italic> in germ cell differentiation and the mechanism by which a <jats:italic>HUWE1<\/jats:italic> single nucleotide polymorphism increases male infertility risk.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>We analyzed <jats:italic>HUWE1<\/jats:italic> single nucleotide polymorphisms in 190 non\u2010obstructive azoospermia patients of Han Chinese descent. We evaluated HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 regulation by retinoic acid receptor alpha using chromatin immunoprecipitation assays, electrophoretic mobility shift assays, and siRNA\u2010mediated <jats:italic>RAR\u03b1<\/jats:italic> knockdown. Using C18\u20104 spermatogonial cells, we determined whether HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 participated in retinoic acid\u2010mediated retinoic acid receptor alpha signaling. We performed luciferase assays, cell counting kit\u20108 assays, immunofluorescence, quantitative real\u2010time polymerase chain reaction, and western blotting. We quantified <jats:italic>HUWE1<\/jats:italic> and retinoic acid receptor alpha in testicular biopsies from non\u2010obstructive azoospermia and obstructive azoospermia patients using quantitative real\u2010time polymerase chain reaction and immunofluorescence.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Three <jats:italic>HUWE1<\/jats:italic> single nucleotide polymorphisms were significantly associated with spermatogenic failure in 190 non\u2010obstructive azoospermia patients; one (rs34492591) was in the <jats:italic>HUWE1<\/jats:italic> promoter. Retinoic acid receptor alpha regulates <jats:italic>HUWE1<\/jats:italic> gene expression by binding to its promoter. HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 participates in retinoic acid\/retinoic acid receptor alpha signaling pathway and regulates the expression of germ cell differentiation genes <jats:italic>STRA8<\/jats:italic> and <jats:italic>SCP3<\/jats:italic> to inhibit cell proliferation and reduce \u03b3H2AX accumulation. Notably, significantly lower levels of <jats:italic>HUWE1<\/jats:italic> and <jats:italic>RAR\u03b1<\/jats:italic> were detected in testicular biopsy samples from non\u2010obstructive azoospermia patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>An <jats:italic>HUWE1<\/jats:italic> promoter single nucleotide polymorphism significantly downregulates its expression in non\u2010obstructive azoospermia patients. Mechanistically, HECT, UBA, and WWE domain\u2010containing E3 ubiquitin protein ligase 1 regulates germ cell differentiation during meiotic prophase through its participation in retinoic acid\/retinoic acid receptor alpha signaling and subsequent modulation of \u03b3H2AX. Taken together, these results strongly suggest that the genetic polymorphisms of <jats:italic>HUWE1<\/jats:italic> are closely related to spermatogenesis and non\u2010obstructive azoospermia pathogenesis.<\/jats:p><\/jats:sec>","DOI":"10.1111\/andr.13474","type":"journal-article","created":{"date-parts":[[2023,6,8]],"date-time":"2023-06-08T20:39:20Z","timestamp":1686256760000},"page":"338-348","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["A novel SNP in <i>HUWE1<\/i> promoter confers increased risk of NOA by affecting the RA\/RAR\u03b1 pathway in Chinese individuals"],"prefix":"10.1111","volume":"12","author":[{"given":"Xudong","family":"Shan","sequence":"first","affiliation":[{"name":"West China School of Basic Medical Sciences &amp; Forensic Medicine Sichuan University Chengdu China"},{"name":"Reproductive &amp; Women\u2010Children Hospital Chengdu University of Traditional Chinese Medicine Chengdu China"},{"name":"School of Medical and Life Sciences Chengdu University of Traditional Chinese Medicine Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3330-373X","authenticated-orcid":false,"given":"Xueguang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics\/Gynecology Key Laboratory of Obstetric Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education West China Second University Hospital Sichuan University Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gelin","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics\/Gynecology Key Laboratory of Obstetric Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education West China Second University Hospital Sichuan University Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1315-1807","authenticated-orcid":false,"given":"Jiao","family":"Lv","sequence":"additional","affiliation":[{"name":"Department of Obstetrics\/Gynecology Key Laboratory of Obstetric Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education West China Second University Hospital Sichuan University Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zixia","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Obstetrics\/Gynecology Key Laboratory of Obstetric Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education West China Second University Hospital Sichuan University Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7049-4569","authenticated-orcid":false,"given":"Chuan","family":"Jiang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics\/Gynecology Key Laboratory of Obstetric Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education West China Second University Hospital Sichuan University Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaohui","family":"Jiang","sequence":"additional","affiliation":[{"name":"Human Sperm Bank West China Second University Hospital Sichuan University Chengdu China"},{"name":"Key Laboratory of Birth Defects and Related Disease of Women and Children (Sichuan University) Ministry of Education Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianxing","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Urology Peking University Third Hospital Peking University Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haocheng","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Urology Peking University Third Hospital Peking University Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6256-2733","authenticated-orcid":false,"given":"Hui","family":"Jiang","sequence":"additional","affiliation":[{"name":"Department of Urology Peking University Third Hospital Peking University Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huanxun","family":"Yue","sequence":"additional","affiliation":[{"name":"Human Sperm Bank West China Second University Hospital Sichuan University Chengdu China"},{"name":"Key Laboratory of Birth Defects and Related Disease of Women and Children (Sichuan University) Ministry of Education Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhengrong","family":"Wang","sequence":"additional","affiliation":[{"name":"West China School of Basic Medical Sciences &amp; 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