{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T03:52:06Z","timestamp":1791345126766,"version":"4.3.1"},"reference-count":0,"publisher":"The Endocrine Society","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Cytochrome b5, encoded by CYB5A, is an essential co-factor for CYP17A1-mediated 17,20-lyase activity. CYB5A deficiency is a rare cause of 46,XY DSD. We report two siblings with 46,XY DSD from a Vietnamese family, presenting with phenotypic female.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Clinical evaluation and hormone profiling were conducted. Whole genome sequencing (WGS) was performed on extracted DNA from both affected siblings and identified variants were classified according to the ACMG\/AMP 2015 guidelines. A minigene RNA splicing assay in HEK293T cells was used to evaluate the functional impact of the CYB5A intronic variant.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The two patients\u2019 hormone profile showed low testosterone and estradiol. WGS identified novel compound heterozygous CYB5A variants: a paternally inherited missense variant (p.Val34Glu) and a maternally inherited deep intronic deletion (c.129\u2009+\u2009862_129\u2009+\u2009863del) which predicted to create the cryptic splice sites by SpliceAI. Minigene assays confirmed the activation of the cryptic splice sites, resulting in an inclusion of a pseudo-exon between exon 1 and 2. This aberrant insertion introduced a premature stop codon, leading to predicted nonsense-mediated decay of the transcript. Both variants were classified as likely pathogenic according to the ACMG\/AMP guideline.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Two novel CYB5A variants established the genetic diagnosis in both siblings, expanding the spectrum of pathogenic CYB5A-related DSD. Our findings show that a deep intronic variant can lead to null protein due to disrupted splicing. These results highlight the value of WGS and functional assays for detecting and validating clinically relevant non-coding variants in DSD.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1210\/clinem\/dgag395","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T11:35:24Z","timestamp":1791286524000},"source":"Crossref","is-referenced-by-count":0,"title":["Biallelic\n                    <i>CYB5A<\/i>\n                    disruptions in 46, XY DSD: Novel missense and deep intronic variants"],"prefix":"10.1210","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-0778-2998","authenticated-orcid":false,"given":"Shirin","family":"Moradifard","sequence":"first","affiliation":[{"name":"Hudson Institute of Medical Research , Melbourne ,","place":["Australia"]},{"name":"Department of Molecular and Translational Science, Monash University , Melbourne ,","place":["Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1838-6100","authenticated-orcid":false,"given":"Thanh Nha Uyen","family":"Le","sequence":"additional","affiliation":[{"name":"Hudson Institute of Medical Research , Melbourne ,","place":["Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nguyen Thu","family":"Ha","sequence":"additional","affiliation":[{"name":"National Children\u2019s Hospital ,","place":["Vietnam"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vu Chi","family":"Dung","sequence":"additional","affiliation":[{"name":"National Children\u2019s Hospital ,","place":["Vietnam"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bui Phuong","family":"Thao","sequence":"additional","affiliation":[{"name":"National Children\u2019s Hospital ,","place":["Vietnam"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2405-1262","authenticated-orcid":false,"given":"Vincent","family":"Harley","sequence":"additional","affiliation":[{"name":"Hudson Institute of Medical Research , Melbourne ,","place":["Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"80","published-online":{"date-parts":[[2026,10,7]]},"container-title":["The Journal of Clinical Endocrinology &amp; Metabolism"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jcem\/advance-article-pdf\/doi\/10.1210\/clinem\/dgag395\/71636760\/dgag395.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jcem\/advance-article-pdf\/doi\/10.1210\/clinem\/dgag395\/71636760\/dgag395.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T03:42:40Z","timestamp":1791344560000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jcem\/advance-article\/doi\/10.1210\/clinem\/dgag395\/8870113"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,7]]},"references-count":0,"URL":"https:\/\/doi.org\/10.1210\/clinem\/dgag395","relation":{},"ISSN":["0021-972X","1945-7197"],"issn-type":[{"value":"0021-972X","type":"print"},{"value":"1945-7197","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,7]]},"article-number":"dgag395"}}