{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T13:32:34Z","timestamp":1786627954107,"version":"build-2736575974"},"reference-count":21,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science Centre in Poland","award":["2020\/37\/B\/NZ5\/00549"],"award-info":[{"award-number":["2020\/37\/B\/NZ5\/00549"]}]},{"award":["2020\/37\/B\/NZ5\/00549"],"award-info":[{"award-number":["2020\/37\/B\/NZ5\/00549"]}],"id":[{"id":"https:\/\/ror.org\/03ha2q922","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>More than 70% of non-obstructive azoospermia (NOA) cases remain idiopathic, and the underlying genetic causes need to be investigated. In this study, we aimed to identify genes and variants associated with the etiology of NOA. Two NOA patients from consanguineous families, with normal karyotypes and no Y-chromosome microdeletions, were selected for whole-genome sequencing (WGS). Candidate variants were validated by Sanger sequencing. Structural protein modeling and assessment of mutation-induced changes in molecular interactions were performed using SWISS-MODEL and AlphaFold. We identified a candidate homozygous missense variant (c.899C&gt;T, p.(Ser300Phe); rs375931805) in SOX30 in one patient; this transcription factor is highly expressed in the testis. In the second patient, a homozygous missense variant (c.1472C&gt;T, p.(Thr491Met); rs746902232) in RNF220 and a hemizygous missense variant (c.2354A&gt;G, p.(Gln785Arg) in the X-linked AKAP4 gene were simultaneously detected. Previous studies have shown that SOX30 knockout in mice leads to meiosis I arrest and impaired spermiogenesis. AKAP4 is exclusively expressed in the testis, and missense variants have been reported in different forms of male infertility. The co-occurrence of variants in AKAP4 and RNF220 may suggest an oligogenic etiology of NOA and contribute to the phenotypic variability associated with AKAP4 variants.<\/jats:p>","DOI":"10.3390\/ijms27167230","type":"journal-article","created":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T12:47:11Z","timestamp":1786625231000},"page":"7230","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Whole-Genome Sequencing Identifies Potentially Causative Variants in SOX30, AKAP4, and RNF220 in Non-Obstructive Azoospermia"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-8474-0601","authenticated-orcid":false,"given":"Razieh","family":"Ebrahimi Askari","sequence":"first","affiliation":[{"name":"Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd 8916978477, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3433-6990","authenticated-orcid":false,"given":"Agnieszka","family":"Malcher","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Polish Academy of Sciences, 60-479 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0867-9305","authenticated-orcid":false,"given":"Fateme","family":"Sefid","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd 8916978477, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9173-5114","authenticated-orcid":false,"given":"Sara","family":"Sadeghzadeh","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd 8916978477, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2284-2264","authenticated-orcid":false,"given":"Rim","family":"Ibrahim","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Polish Academy of Sciences, 60-479 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5562-0114","authenticated-orcid":false,"given":"Krzysztof","family":"Pastuszak","sequence":"additional","affiliation":[{"name":"Department of Algorithms and System Modelling, Gdansk University of Technology, 80-233 Gdansk, Poland"},{"name":"Department of Translational Oncology, Medical University of Gdansk, 80-210 Gdansk, Poland"},{"name":"Centre of Biostatistics and Bioinformatics, Medical University of Gdansk, 80-210 Gdansk, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aashish","family":"Srivastava","sequence":"additional","affiliation":[{"name":"Genome Core Facility, Department of Clinical Science, Haukeland University Hospital, 5021 Bergen, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyed","family":"Mehdi Kalantar","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd 8916978477, Iran"},{"name":"Abortion Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd 8916877391, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Serajoddin","family":"Vahidi","sequence":"additional","affiliation":[{"name":"Andrology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd 8916877391, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nasrin","family":"Ghasemi","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd 8916978477, Iran"},{"name":"Abortion Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd 8916877391, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3275-3245","authenticated-orcid":false,"given":"Maciej","family":"Kurpisz","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics, Polish Academy of Sciences, 60-479 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"15","DOI":"10.6061\/clinics\/2013(Sup01)03","article-title":"The epidemiology and etiology of azoospermia","volume":"68","author":"Cocuzza","year":"2013","journal-title":"Clinics"},{"key":"ref_2","first-page":"17","article-title":"Genetic evaluation of male infertility","volume":"3","author":"Wosnitzer","year":"2014","journal-title":"Transl. Androl. Urol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Cioppi, F., Rosta, V., and Krausz, C. (2021). Genetics of azoospermia. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22063264"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1111\/andr.13269","article-title":"Whole-genome sequencing identifies new candidate genes for nonobstructive azoospermia","volume":"10","author":"Malcher","year":"2022","journal-title":"Andrology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"101048","DOI":"10.1016\/j.repbio.2025.101048","article-title":"Summarizing the human genes and their variants causative of non-obstructive azoospermia uncovered using whole genome\/exome sequencing","volume":"25","author":"Ibrahim","year":"2025","journal-title":"Reprod. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zhao, Y., Lockhart, S., Liu, J., Li, X., Cortes, A., Hua, X., Gardner, E., Kentistou, A., Lo, Y., and Davitte, J. (2024). Population scale whole genome sequencing provides novel insights into cardiometabolic health. medRxiv.","DOI":"10.1101\/2024.05.27.24307970"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"dev164723","DOI":"10.1242\/dev.164723","article-title":"The transcription factor SOX30 is a key regulator of mouse spermiogenesis","volume":"145","author":"Zhang","year":"2018","journal-title":"Development"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"dev164855","DOI":"10.1242\/dev.164855","article-title":"Sox30 initiates transcription of haploid genes during late meiosis and spermiogenesis in mouse testes","volume":"145","author":"Bai","year":"2018","journal-title":"Development"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1038\/s41419-025-07442-1","article-title":"Single-cell RNA-seq and pathological phenotype reveal the functional atlas and precise roles of Sox30 in testicular cell development and differentiation","volume":"16","author":"Xie","year":"2025","journal-title":"Cell Death Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.omtn.2019.10.038","article-title":"Epigenetic inactivation of SOX30 is associated with male infertility and offers a therapy target for non-obstructive azoospermia","volume":"19","author":"Han","year":"2020","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1007\/s10528-025-11050-1","article-title":"Exploring miR-34a, miR-449, and ADAM2\/ADAM7 expressions as potential biomarkers in male infertility: A combined in silico and experimental approach","volume":"64","author":"Ghodrati","year":"2025","journal-title":"Biochem. Genet."},{"key":"ref_12","first-page":"1000406","article-title":"SOX30 might not be associated with Sertoli cell-only syndrome in azoospermic Japanese men","volume":"8","author":"Miyamoto","year":"2020","journal-title":"Integr. Mol. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1093\/biolre\/ioaf258","article-title":"Mutations of SOX30 are identified in azoospermic human patients and reduce sperm in mice","volume":"114","author":"Ren","year":"2026","journal-title":"Biol. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.stem.2018.08.007","article-title":"Single-cell RNA sequencing analysis reveals sequential cell fate transition during human spermatogenesis","volume":"23","author":"Wang","year":"2018","journal-title":"Cell Stem Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1002\/jemt.10320","article-title":"Fibrous sheath of mammalian spermatozoa","volume":"61","author":"Eddy","year":"2003","journal-title":"Microsc. Res. Tech."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1002\/mrd.23529","article-title":"A new hemizygous missense mutation, c.454T>C (p.S152P), in AKAP4 gene is associated with asthenozoospermia","volume":"88","author":"Gu","year":"2021","journal-title":"Mol. Reprod. Dev."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e1463","DOI":"10.1002\/ctm2.1463","article-title":"A pathogenic AKAP4 variant, p.R429H, causes male in\/subfertility in humans and mice","volume":"13","author":"Wei","year":"2023","journal-title":"Clin. Transl. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"189","DOI":"10.4103\/jhrs.JHRS_22_19","article-title":"The effect of sperm DNA fragmentation index on the outcome of intrauterine insemination and intracytoplasmic sperm injection","volume":"12","author":"Siddhartha","year":"2019","journal-title":"J. Hum. Reprod. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"25","DOI":"10.4103\/aja202329","article-title":"Decreased AKAP4\/PKA signaling pathway in high DFI sperm affects sperm capacitation","volume":"26","author":"Zhang","year":"2024","journal-title":"Asian J. Androl."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.gene.2018.11.074","article-title":"A homozygous RNF220 mutation leads to male infertility with small-headed sperm","volume":"688","author":"Jiang","year":"2019","journal-title":"Gene"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1111\/andr.12378","article-title":"Next-generation sequencing for patients with non-obstructive azoospermia: Implications for significant roles of monogenic\/oligogenic mutations","volume":"5","author":"Nakamura","year":"2017","journal-title":"Andrology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/16\/7230\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T12:47:33Z","timestamp":1786625253000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/16\/7230"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,13]]},"references-count":21,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27167230"],"URL":"https:\/\/doi.org\/10.3390\/ijms27167230","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,13]]}}}