{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T18:56:33Z","timestamp":1757616993957,"version":"3.44.0"},"reference-count":22,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,10,2]],"date-time":"2024-10-02T00:00:00Z","timestamp":1727827200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,10,2]],"date-time":"2024-10-02T00:00:00Z","timestamp":1727827200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100008131","name":"Rheinische Friedrich-Wilhelms-Universit\u00e4t Bonn","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100008131","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>\n            <jats:italic>Isl1<\/jats:italic> has been described as an embryonic master control gene expressed in the pericloacal mesenchyme. Deletion of <jats:italic>Isl1<\/jats:italic> from the genital mesenchyme in mice leads to an ectopic urethral opening and epispadias-like phenotype. Using genome wide association methods, we identified <jats:italic>ISL1<\/jats:italic> as the key susceptibility gene for classic bladder exstrophy (CBE), comprising epispadias and exstrophy of the urinary bladder. The most significant marker (rs6874700) identified in our recent GWAS meta-analysis achieved a <jats:italic>p<\/jats:italic> value of 1.48\u2009\u00d7\u200910<jats:sup>\u2212\u200924<\/jats:sup> within the <jats:italic>ISL1<\/jats:italic> region. In silico analysis of rs6874700 and all other genome-wide significant markers in Linkage Disequilibrium (LD) with rs6874700 (D\u2019 = 1.0; R<jats:sup>2<\/jats:sup>\u2009&gt;\u20090.90) revealed marker rs2303751 (<jats:italic>p<\/jats:italic> value 8.12\u2009\u00d7\u200910<jats:sup>\u2212\u200920<\/jats:sup>) as the marker with the highest regulatory effect predicted. Here, we describe a novel 1.2\u00a0kb intragenic promoter residing between 6.2 and 7.4\u00a0kb downstream of the <jats:italic>ISL1<\/jats:italic> transcription starting site, which is located in the reverse DNA strand and harbors a binding site for EZH2 at the exact region of marker rs2303751. We show, that EZH2 silencing in HEK cells reduces <jats:italic>ISL1<\/jats:italic> expression. We show that <jats:italic>ezh2<\/jats:italic>\n            <jats:sup>\u2212\/\u2212<\/jats:sup> knockout (KO) zebrafish larvae display tissues specificity of ISL1 regulation with reduced expression of Isl1 in the pronephric region of zebrafish larvae. In addition, a shorter and malformed nephric duct is observed in <jats:italic>ezh2<\/jats:italic>\n            <jats:sup>\u2212\/\u2212<\/jats:sup> ko zebrafish <jats:italic>Tg(wt1\u00df:eGFP)<\/jats:italic> reporter lines. Our study shows, that Ezh2 is a key regulator of <jats:italic>Isl1<\/jats:italic> during urinary tract formation and suggests tissue specific <jats:italic>ISL1<\/jats:italic> dysregulation as an underlying mechanism for CBE formation.<\/jats:p>","DOI":"10.1038\/s41598-024-74303-w","type":"journal-article","created":{"date-parts":[[2024,10,2]],"date-time":"2024-10-02T10:02:10Z","timestamp":1727863330000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["EZH2 specifically regulates ISL1 during embryonic urinary tract formation"],"prefix":"10.1038","volume":"14","author":[{"given":"Enrico","family":"Mingardo","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeshurun C.","family":"Kalanithy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriel","family":"Dworschak","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nina","family":"Ishorst","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00d6znur","family":"Yilmaz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tobias","family":"Lindenberg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ronja","family":"Hollstein","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tim","family":"Felger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pierre-Olivier","family":"Angrand","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heiko","family":"Reutter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benjamin","family":"Odermatt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,10,2]]},"reference":[{"key":"74303_CR1","doi-asserted-by":"publisher","unstructured":"Suzuki, K. et al. Reduced BMP signaling results in hindlimb fusion with lethal pelvic\/urogenital organ aplasia: a new mouse model of sirenomelia. PloS One. 7 (9). https:\/\/doi.org\/10.1371\/JOURNAL.PONE.0043453 (2012).","DOI":"10.1371\/JOURNAL.PONE.0043453"},{"key":"74303_CR2","doi-asserted-by":"publisher","unstructured":"Ching, S. T. et al. Isl1 mediates mesenchymal expansion in the developing external genitalia via regulation of Bmp4, Fgf10 and Wnt5a. Hum. Mol. Genet.  27 (1). https:\/\/doi.org\/10.1093\/hmg\/ddx388 (2018).","DOI":"10.1093\/hmg\/ddx388"},{"issue":"3","key":"74303_CR3","doi-asserted-by":"publisher","first-page":"e1005024","DOI":"10.1371\/JOURNAL.PGEN.1005024","volume":"11","author":"M Draaken","year":"2015","unstructured":"Draaken, M. et al. Genome-wide Association study and Meta-analysis identify ISL1 as genome-wide significant susceptibility gene for bladder exstrophy. PLoS Genet.  11 (3), e1005024. https:\/\/doi.org\/10.1371\/JOURNAL.PGEN.1005024 (2015).","journal-title":"PLoS Genet."},{"key":"74303_CR4","doi-asserted-by":"publisher","unstructured":"Beaman, G. M., Cervellione, R. M., Keene, D., Reutter, H. & Newman, W. G. The genomic architecture of bladder exstrophy epispadias complex. In Genes (Vol. 12, Issue 8). (2021). https:\/\/doi.org\/10.3390\/genes12081149","DOI":"10.3390\/genes12081149"},{"issue":"1","key":"74303_CR5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s42003-022-04092-3","volume":"2022 5:1","author":"E Mingardo","year":"2022","unstructured":"Mingardo, E. et al. A genome-wide association study with tissue transcriptomics identifies genetic drivers for classic bladder exstrophy. Commun. Biology. 2022 5:1 (1), 1\u201311. https:\/\/doi.org\/10.1038\/s42003-022-04092-3 (2022). 5.","journal-title":"Commun. Biology"},{"key":"74303_CR6","doi-asserted-by":"publisher","unstructured":"Kim, J. et al. Polycomb- and methylation-independent roles of EZH2 as a transcription activator. Cell. Rep.  25 (10). https:\/\/doi.org\/10.1016\/j.celrep.2018.11.035 (2018).","DOI":"10.1016\/j.celrep.2018.11.035"},{"key":"74303_CR7","doi-asserted-by":"publisher","unstructured":"Dupret, B. et al. The histone lysine methyltransferase Ezh2 is required for maintenance of the intestine integrity and for caudal fin regeneration in zebrafish. Biochim. et Biophys. Acta - Gene Regul. 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Cell Death Dis.  10 (10). https:\/\/doi.org\/10.1038\/s41419-019-1952-z (2019).","DOI":"10.1038\/s41419-019-1952-z"},{"key":"74303_CR14","doi-asserted-by":"publisher","unstructured":"Arkani, S. et al. Evaluation of the ISL1 gene in the pathogenesis of bladder exstrophy in a Swedish cohort. Hum. Genome Variation. 5https:\/\/doi.org\/10.1038\/hgv.2018.9 (2018).","DOI":"10.1038\/hgv.2018.9"},{"issue":"2","key":"74303_CR15","doi-asserted-by":"publisher","first-page":"378","DOI":"10.1002\/AJMG.A.63031","volume":"191","author":"A Nordenskj\u00f6ld","year":"2023","unstructured":"Nordenskj\u00f6ld, A. et al. Copy number variants suggest different molecular pathways for the pathogenesis of bladder exstrophy. Am. J. Med. Genet. Part. A. 191 (2), 378\u2013390. https:\/\/doi.org\/10.1002\/AJMG.A.63031 (2023).","journal-title":"Am. J. Med. Genet. Part. A"},{"key":"74303_CR16","doi-asserted-by":"publisher","unstructured":"Bernstein, B. E. et al. A bivalent chromatin structure Marks Key Developmental genes in embryonic stem cells. Cell. 125 (2). https:\/\/doi.org\/10.1016\/j.cell.2006.02.041 (2006).","DOI":"10.1016\/j.cell.2006.02.041"},{"issue":"5909","key":"74303_CR17","doi-asserted-by":"publisher","first-page":"1849","DOI":"10.1126\/SCIENCE.1162253","volume":"322","author":"AC Seila","year":"2008","unstructured":"Seila, A. C. et al. Divergent transcription from active promoters. Sci. (New York N Y). 322 (5909), 1849. https:\/\/doi.org\/10.1126\/SCIENCE.1162253 (2008).","journal-title":"Sci. (New York N Y)"},{"key":"74303_CR18","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/J.DIFF.2018.08.004","volume":"103","author":"A Liaw","year":"2018","unstructured":"Liaw, A. et al. Development of the human bladder and ureterovesical junction. Differ. Res. Biol. Divers.  103, 66\u201373. https:\/\/doi.org\/10.1016\/J.DIFF.2018.08.004 (2018).","journal-title":"Differ. Res. Biol. 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BMC Genom.  14 (1), 1\u201320. https:\/\/doi.org\/10.1186\/1471-2164-14-914\/FIGURES\/7 (2013).","journal-title":"BMC Genom."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-74303-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-74303-w","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-74303-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T23:04:40Z","timestamp":1757113480000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-74303-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,2]]},"references-count":22,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["74303"],"URL":"https:\/\/doi.org\/10.1038\/s41598-024-74303-w","relation":{},"ISSN":["2045-2322"],"issn-type":[{"type":"electronic","value":"2045-2322"}],"subject":[],"published":{"date-parts":[[2024,10,2]]},"assertion":[{"value":"24 June 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 September 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 October 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 July 2025","order":4,"name":"change_date","label":"Change Date","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Update","order":5,"name":"change_type","label":"Change Type","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"This article has been updated to amend the license information.","order":6,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no conflict of interest.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"All authors have seen and approved the manuscript. This manuscript and its data have not been accepted or published elsewhere.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Author Approvals"}},{"value":"Animal husbandry and experimental setups were in accordance with European Legislation for the Protection of Animals used for Scientific Purposes (Directive 2010\/62\/EU). National law exempts all zebrafish experiments performed in larval stages up to five dpf before independent feeding from ethical approval. All experiments performed in zebrafish larvae from 0 to 5 dpf do not require any Ethics committee approval. All experiments are in accordance with ARRIVE guidelines.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics Declaration"}}],"article-number":"22909"}}