{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T04:55:30Z","timestamp":1783486530235,"version":"3.55.0"},"reference-count":47,"publisher":"Public Library of Science (PLoS)","issue":"1","license":[{"start":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T00:00:00Z","timestamp":1735862400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000069","name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases","doi-asserted-by":"publisher","award":["AR076747"],"award-info":[{"award-number":["AR076747"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000069","name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases","doi-asserted-by":"publisher","award":["AR072987"],"award-info":[{"award-number":["AR072987"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Lateral Meningocele Syndrome (LMS), a disorder associated with <jats:italic>NOTCH3<\/jats:italic> pathogenic variants, presents with neurological, craniofacial and skeletal abnormalities. Mouse models of the disease exhibit osteopenia that is ameliorated by the administration of Notch3 antisense oligonucleotides (ASO) targeting either <jats:italic>Notch3<\/jats:italic> or the <jats:italic>Notch3<\/jats:italic> mutation. To determine the consequences of LMS pathogenic variants in human cells and whether they can be targeted by ASOs, induced pluripotent NCRM1 and NCRM5 stem (iPS) cells harboring a <jats:italic>NOTCH3<\/jats:italic><jats:sup><jats:italic>6692-93insC<\/jats:italic><\/jats:sup> insertion were created. Parental iPSCs, <jats:italic>NOTCH3<\/jats:italic><jats:sup><jats:italic>6692-93insC<\/jats:italic><\/jats:sup> and isogenic controls, free of chromosomal aberrations as determined by human CytoSNP850 array, were cultured under conditions of neural crest, mesenchymal and osteogenic cell differentiation. The expected cell phenotype was confirmed by surface markers and a decline in <jats:italic>OCT3\/4<\/jats:italic> and <jats:italic>NANOG<\/jats:italic> mRNA. <jats:italic>NOTCH3<\/jats:italic><jats:sup><jats:italic>6692-93insC<\/jats:italic><\/jats:sup> cells displayed enhanced expression of Notch target genes <jats:italic>HES1<\/jats:italic>, <jats:italic>HEY1<\/jats:italic>, <jats:italic>2<\/jats:italic> and <jats:italic>L<\/jats:italic> demonstrating a NOTCH3 gain-of-function. There was enhanced osteogenesis in <jats:italic>NOTCH3<\/jats:italic><jats:sup><jats:italic>6692-93insC<\/jats:italic><\/jats:sup> cells as evidenced by increased mineralized nodule formation and <jats:italic>ALPL<\/jats:italic>, <jats:italic>BGLAP<\/jats:italic> and <jats:italic>BSP<\/jats:italic> expression. ASOs targeting <jats:italic>NOTCH3<\/jats:italic> decreased both <jats:italic>NOTCH3<\/jats:italic> wild type and <jats:italic>NOTCH3<\/jats:italic><jats:sup><jats:italic>6692-93insC<\/jats:italic><\/jats:sup> mutant mRNA by 40% in mesenchymal and 90% in osteogenic cells. ASOs targeting the <jats:italic>NOTCH3<\/jats:italic> insertion decreased <jats:italic>NOTCH3<\/jats:italic><jats:sup><jats:italic>6692-93insC<\/jats:italic><\/jats:sup> by 70\u201380% in mesenchymal cells and by 45\u201355% in osteogenic cells and <jats:italic>NOTCH3<\/jats:italic> mRNA by 15\u201330% and 20\u201340%, respectively. In conclusion, a <jats:italic>NOTCH3<\/jats:italic> pathogenic variant causes a modest increase in osteoblastogenesis in human iPS cells <jats:italic>in vitro<\/jats:italic> and NOTCH3 and NOTCH3 mutant specific ASOs downregulate <jats:italic>NOTCH3<\/jats:italic> transcripts associated with LMS.<\/jats:p>","DOI":"10.1371\/journal.pone.0316644","type":"journal-article","created":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T18:37:11Z","timestamp":1735929431000},"page":"e0316644","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["A NOTCH3 pathogenic variant influences osteogenesis and can be targeted by antisense oligonucleotides in induced pluripotent stem cells"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9154-952X","authenticated-orcid":true,"given":"Ernesto","family":"Canalis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jungeun","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren","family":"Schilling","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paymaan","family":"Jafar-nejad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6464-9584","authenticated-orcid":true,"given":"Michele","family":"Carrer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,1,3]]},"reference":[{"issue":"1","key":"pone.0316644.ref001","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0022-3476(77)80763-4","article-title":"Familial osteosclerosis with abnormalities of the nervous system and meninges","volume":"90","author":"RA Lehman","year":"1977","journal-title":"J Pediatr"},{"issue":"3","key":"pone.0316644.ref002","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1002\/(SICI)1096-8628(19970613)70:3<229::AID-AJMG4>3.0.CO;2-R","article-title":"Lateral meningocele syndrome: three new patients and review of the literature","volume":"70","author":"KW Gripp","year":"1997","journal-title":"Am J Med Genet"},{"issue":"3","key":"pone.0316644.ref003","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1002\/ajmg.a.33510","article-title":"Hajdu-Cheney syndrome with severe dural ectasia","volume":"155A","author":"K Avela","year":"2011","journal-title":"Am J Med Genet A"},{"issue":"2","key":"pone.0316644.ref004","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1002\/ajmg.a.36863","article-title":"Truncating mutations in the last exon of NOTCH3 cause lateral meningocele syndrome","volume":"167A","author":"KW Gripp","year":"2015","journal-title":"Am J Med Genet A"},{"issue":"7","key":"pone.0316644.ref005","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1002\/ajmg.a.34075","article-title":"Lateral meningocele syndrome and Hajdu-Cheney syndrome: different disorders with overlapping phenotypes","volume":"155A","author":"KW Gripp","year":"2011","journal-title":"Am J Med Genet A"},{"issue":"12","key":"pone.0316644.ref006","doi-asserted-by":"crossref","first-page":"2611","DOI":"10.1007\/s00198-018-4694-3","article-title":"Notch in skeletal physiology and disease.","volume":"29","author":"E. 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