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Recent advances in genetic medicine have led to a better understanding of the genotype\u2013phenotype correlation in <jats:italic>WT1<\/jats:italic>-related diseases, particularly missense variants in exons 8 or 9 that lead to a wide range of severities. Exonic variants can lead to splicing abnormalities in rare diseases. No reports have investigated exonic variants in <jats:italic>WT1<\/jats:italic> that cause aberrant splicing. We examined whether exonic variants in <jats:italic>WT1<\/jats:italic> exon 8 or 9 cause splicing abnormalities and affect disease severity.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>We selected nine rare missense variants in exon 8 or 9 in <jats:italic>WT1<\/jats:italic> outside the DNA binding domain and C2H2 sites from the Human Gene Variant Database Professional that unexpectedly present severe phenotypes. We conducted functional splicing assays using hybrid minigenes for the nine variants containing exon 8 and 9 and surrounding sequences. Minigene vectors were transfected into cultured cells, and mRNA was analyzed. In silico analysis was performed.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Splicing assays revealed that one of the nine variants caused aberrant splicing, with exon 8 skipping. One previously reported case with this variant showed particularly severe phenotype, progressing to kidney failure within 3\u00a0months.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>One <jats:italic>WT1<\/jats:italic> variant in exon 8 or 9 disrupted the splice site, leading to aberrant splicing in vitro and potentially contributing to an unexpectedly severe phenotype for a missense variant outside the DNA binding domain and C2H2 sites. In vitro splicing assays may help clarify the genotype\u2013phenotype correlation in <jats:italic>WT1<\/jats:italic>-related disorders, especially for variants outside canonical functional domains.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s10157-025-02715-7","type":"journal-article","created":{"date-parts":[[2025,6,10]],"date-time":"2025-06-10T04:49:28Z","timestamp":1749530968000},"page":"1489-1495","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Potential involvement of abnormal splicing in severe WT1-related disorders"],"prefix":"10.1007","volume":"29","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4946-1345","authenticated-orcid":false,"given":"China","family":"Nagano","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masafumi","family":"Matuso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuta","family":"Inoki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuta","family":"Ichikawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chika","family":"Ueda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideaki","family":"Kitakado","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nana","family":"Sakakibara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomoko","family":"Horinouchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohiko","family":"Yamamura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shingo","family":"Ishimori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kandai","family":"Nozu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,6,10]]},"reference":[{"issue":"9","key":"2715_CR1","doi-asserted-by":"publisher","first-page":"2601","DOI":"10.1007\/s00467-024-06302-y","volume":"39","author":"M Lopez-Gonzalez","year":"2024","unstructured":"Lopez-Gonzalez M, Ariceta G. 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Ltd., Toa Eiyo Ltd., Zenyaku Kogyo Co. Ltd., and Taisho Pharmaceutical Co. Ltd. Kandai Nozu received lecture fees from Ono Pharma, Astellas Pharma, Novo Nordisk Pharma, Alexion Pharma, Sumitomo Pharma, Sanofi, Otsuka Pharma, Daiichi Sankyo, and Miyarisan. Kandai Nozu received a grant from Zenyaku Kogyo and Torii Co. Ltd. Tomoko Horinouchi received research funding from Otsuka Pharmaceutical Co., Ltd., Sysmex Corporation, and Zenyaku Kogyo Co., Ltd. The other authors have nothing to disclose.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"All procedures performed in studies involving human participants were in accordance with the ethical standards of the Institutional Review Board of Kobe University Graduate School of Medicine (IRB approval number 301) and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all participants included in the study.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}},{"value":"Informed consent was obtained from all individual participants included in the study.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Informed consent"}}]}}