{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T00:21:00Z","timestamp":1768522860446,"version":"3.49.0"},"posted":{"date-parts":[[2025,7,22]]},"group-title":"Developmental Biology","reference-count":37,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T00:00:00Z","timestamp":1753142400000},"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":"crossref","award":["R56AR080718"],"award-info":[{"award-number":["R56AR080718"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"crossref","award":["R03DE031388"],"award-info":[{"award-number":["R03DE031388"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000867","name":"Robert Wood Johnson Foundation","doi-asserted-by":"crossref","award":["79430"],"award-info":[{"award-number":["79430"]}],"id":[{"id":"10.13039\/100000867","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,7,22]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Introduction<\/jats:title>\n                  <jats:p>\n                    Robinow syndrome has characteristic craniofacial and dental features and can be caused by gain- or loss-of-function variants in Wnt family member 5A (\n                    <jats:italic>WNT5A<\/jats:italic>\n                    ) non-canonical signaling. The craniofacial and dental manifestation of Robinow syndrome is heterogenous, as is the effect of altered\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    in animal models. The relationship between\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    and craniofacial and dental phenotypes is not fully understood.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    To investigate the role of\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    in craniofacial and dental development, we utilized a\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    conditional loss-of-function (LOF:\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    <jats:sup>\n                      <jats:italic>fl\/fl<\/jats:italic>\n                    <\/jats:sup>\n                    <jats:italic>;Ctsk<\/jats:italic>\n                    <jats:sup>\n                      <jats:italic>cre<\/jats:italic>\n                    <\/jats:sup>\n                    ) and a\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    conditional gain-of-function (GOF:\n                    <jats:italic>Rosa26-LSL-Wnt5a;Ctsk<\/jats:italic>\n                    <jats:sup>\n                      <jats:italic>cre<\/jats:italic>\n                    <\/jats:sup>\n                    <jats:italic>)<\/jats:italic>\n                    model to determine the effect of both LOF and GOF of\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    in bone cells during craniofacial and dental development. Postnatal day 10 conditional LOF\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    , GOF\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    , and control skulls were scanned by micro-computed tomography and assessed using traditional and geometric morphometrics. Mandibular bone apoptosis was further assessed by TUNEL staining.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Conditional\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    LOF resulted in midface hypoplasia, increased maxillary intermolar width, increased rostral basisphenoid width, and delayed molar eruption.\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    LOF mandibles did not have altered bone mineral density or bone microarchitecture unlike our previous study examining\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    LOF femurs. In contrast, conditional\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    GOF results in macrocephaly, shortened hard palate, increased zygomatic length, micrognathia, and mandibular process morphology changes. The micrognathia and mandibular process morphology changes in the\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    GOF mice were not due to increased apoptosis. A partially penetrant snout deviation was present in both the\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    LOF and GOF mice.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>\n                    Craniofacial and dental phenotype differed between mice with conditional GOF and LOF of\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    , consistent with the craniofacial phenotype heterogeneity in\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    -associated Robinow syndrome. We detected tooth eruption delay, mandibular condyle dysmorphology, and facial asymmetry in mice with altered\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    that have not been previously reported in patients. Our data suggest precise regulation of\n                    <jats:italic>Wnt5a<\/jats:italic>\n                    is essential for proper craniofacial and dental development.\n                  <\/jats:p>\n                  <jats:fig id=\"ufig1\" position=\"float\" fig-type=\"figure\" orientation=\"portrait\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"665966v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.1101\/2025.07.21.665966","type":"posted-content","created":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T18:50:11Z","timestamp":1753210211000},"source":"Crossref","is-referenced-by-count":0,"title":["Wnt5a gain- and loss-of-function present distinctly in craniofacial bone"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2003-4625","authenticated-orcid":false,"given":"Claire J.","family":"Houchen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Portia","family":"Hahn Leat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9109-2839","authenticated-orcid":false,"given":"Cassandra","family":"Delich","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jocelyn","family":"Vang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"Haggard","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6839-1173","authenticated-orcid":false,"given":"Joseph L.","family":"Roberts","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3322-281X","authenticated-orcid":false,"given":"Hicham","family":"Drissi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8698-7808","authenticated-orcid":false,"given":"Erin E.","family":"Bumann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025072418001348000_2025.07.21.665966v1.1","doi-asserted-by":"publisher","DOI":"10.1136\/jmg.23.4.350"},{"issue":"6","key":"2025072418001348000_2025.07.21.665966v1.2","first-page":"00223","article-title":"An osteoblast origin for craniofacial dysplasia in neurofibromatosis type i","volume":"6","year":"2017","journal-title":"J Dent Health Oral Disord Ther"},{"issue":"2","key":"2025072418001348000_2025.07.21.665966v1.3","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.3892\/mmr.2015.4642","article-title":"Upregulation of the expression of wnt5a promotes the proliferation of pancreatic cancer cells in vitro and in a nude mouse model","volume":"13","year":"2016","journal-title":"Molecular Medicine Reports"},{"issue":"2","key":"2025072418001348000_2025.07.21.665966v1.4","doi-asserted-by":"crossref","first-page":"17","DOI":"10.3390\/jdb10020017","article-title":"Genetic interaction of thm2 and thm1 shapes postnatal craniofacial bone","volume":"10","year":"2022","journal-title":"Journal of Developmental Biology"},{"issue":"1","key":"2025072418001348000_2025.07.21.665966v1.5","first-page":"e10706","article-title":"Visualizing cathepsin k cre expression at the single cell level with gfp reporters","volume":"7","year":"2023","journal-title":"Journal of Bone and Mineral Research Plus"},{"issue":"1","key":"2025072418001348000_2025.07.21.665966v1.6","doi-asserted-by":"crossref","first-page":"e1004145","DOI":"10.1371\/journal.pgen.1004145","article-title":"Wnt7b promotes bone formation in part through mtorc1","volume":"10","year":"2014","journal-title":"PLoS genetics"},{"key":"2025072418001348000_2025.07.21.665966v1.7","doi-asserted-by":"publisher","DOI":"10.1002\/gene.20041"},{"issue":"12","key":"2025072418001348000_2025.07.21.665966v1.8","doi-asserted-by":"crossref","first-page":"3606","DOI":"10.1002\/ajmg.a.61986","article-title":"Craniofacial phenotypes associated with robinow syndrome","volume":"185","year":"2021","journal-title":"American Journal of Medical Genetics Part A"},{"key":"2025072418001348000_2025.07.21.665966v1.9","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0554-8"},{"issue":"9","key":"2025072418001348000_2025.07.21.665966v1.10","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1177\/00220345221084200","article-title":"Identification of periosteal osteogenic progenitors in jawbone","volume":"101","year":"2022","journal-title":"Journal of Dental Research"},{"key":"2025072418001348000_2025.07.21.665966v1.11","doi-asserted-by":"crossref","unstructured":"Hassan MG , Kaler H , Zhang B , Cox TC , Young N , Jheon AH . 2020. 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