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Known disease genes explain about 20% of all cases often correlated with primary ciliary dyskinesia (PCD). We aimed to identify disease genes beyond PCD-related aetiologies using exome sequencing in 16 case-parent trios followed by exome survey in 2,109 individuals with situs inversus totalis, heterotaxy, or isolated CHD. We identified six different variants in\n                    <jats:italic>FAM222B<\/jats:italic>\n                    , previously discussed as a candidate gene for cerebral cavernous malformations\n                    <jats:italic>.<\/jats:italic>\n                    The variant c.899G\u2009&gt;\u2009A (p.300Arg\u2009&gt;\u2009His) was found de novo in two unrelated families. FAM222B has been described as a substrate of Nemo-like kinase (NLK), associated with left\u2013right body axis determination. Structural modelling suggests a function of FAM222B Arg300 in NLK recognition. We investigated whole-mount in situ hybridization (WISH) expression pattern and genomic context of the zebrafish (zf)\n                    <jats:italic>FAM222B<\/jats:italic>\n                    homologues,\n                    <jats:italic>fam222ba\/bb,<\/jats:italic>\n                    and\n                    <jats:italic>fam222aa<\/jats:italic>\n                    . Using a double-knockout (dd-KO)\n                    <jats:italic>fam222ba\/bb<\/jats:italic>\n                    zf line, we identified aberrant cardiac looping in these larvae and enlarged atrium and ventricle in adult zf. We further tested the c.899G\u2009&gt;\u2009A variant using human mRNA injections in\n                    <jats:italic>Tg(kdrl:EGFP)<\/jats:italic>\n                    wildtype (wt) reporter-zf, which led to perturbed cardiogenesis. Together, we propose\n                    <jats:italic>FAM222B<\/jats:italic>\n                    as a novel candidate gene for cardiovascular laterality defects.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-64853-6","type":"journal-article","created":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T05:05:41Z","timestamp":1787375141000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Molecular characterization of FAM222B as a novel disease gene for dominant cardiovascular laterality defects"],"prefix":"10.1038","volume":"16","author":[{"given":"Nina","family":"Reitz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Lambertz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00d6znur","family":"Yilmaz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobias T.","family":"Lindenberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khadija","family":"Channab","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birgit","family":"Rau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefanie","family":"Ramrath","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrico","family":"Mingardo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanna","family":"Sch\u00f6pper","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bettina C.","family":"Kirchmaier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias","family":"Geyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel C.","family":"Dworschak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes","family":"Breuer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"M\u00fcller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birthe","family":"Schaidinger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alina C.","family":"Hilger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Hoefele","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Korbinian M.","family":"Riedhammer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc-Phillip","family":"Hitz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregor","family":"Dombrowsky","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hashim","family":"Abdul-Khaliq","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrike M. 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DNA isolation and exome sequencing of affected case-parent trios (HET15 and HET17) was carried out consenting to the IRB ethical approval and was authorized by the local ethics committee of the medical faculty of the University of Bonn (Lfd Nr. 141\/15), as previously published for other laterality defects associated disease genes by Breuer et al. in 2022\n                      11\n                      . In addition to that, written informed consent from all participating families were obtained. Given the potential for subclinical phenotypic expression of laterality defects among relatives of the index case, all included family members underwent comprehensive clinical assessments and targeted ultrasound evaluations performed by two paediatric cardiologists to ascertain their affection status\n                      11\n                      . Zf experiments were performed in larval stages from 0 to 5 days post fertilization before independent feeding and are therefore exempted from ethical approval. All experiments were conducted in accordance with the ARRIVE guidelines.","order":2,"name":"Ethics","label":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"26359"}}