{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T03:02:16Z","timestamp":1790391736829,"version":"4.1.0"},"reference-count":41,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2024,4,26]],"date-time":"2024-04-26T00:00:00Z","timestamp":1714089600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82271186"],"award-info":[{"award-number":["82271186"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81974143"],"award-info":[{"award-number":["81974143"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2022-PUMCH-C-029"],"award-info":[{"award-number":["2022-PUMCH-C-029"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National High Level Hospital Clinical Research Funding","award":["82271186"],"award-info":[{"award-number":["82271186"]}]},{"name":"National High Level Hospital Clinical Research Funding","award":["81974143"],"award-info":[{"award-number":["81974143"]}]},{"name":"National High Level Hospital Clinical Research Funding","award":["2022-PUMCH-C-029"],"award-info":[{"award-number":["2022-PUMCH-C-029"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hemifacial microsomia (HFM) is a rare congenital genetic syndrome primarily affecting the first and second pharyngeal arches, leading to defects in the mandible, external ear, and middle ear. The pathogenic genes remain largely unidentified. Whole-exome sequencing (WES) was conducted on 12 HFM probands and their unaffected biological parents. Predictive structural analysis of the target gene was conducted using PSIPRED (v3.3) and SWISS-MODEL, while STRING facilitated protein-to-protein interaction predictions. CRISPR\/Cas9 was applied for gene knockout in zebrafish. In situ hybridization (ISH) was employed to examine the spatiotemporal expression of the target gene and neural crest cell (NCC) markers. Immunofluorescence with PH3 and TUNEL assays were used to assess cell proliferation and apoptosis. RNA sequencing was performed on mutant and control embryos, with rescue experiments involving target mRNA injections and specific gene knockouts. CDC27 was identified as a novel candidate gene for HFM, with four nonsynonymous de novo variants detected in three unrelated probands. Structural predictions indicated significant alterations in the secondary and tertiary structures of CDC27. cdc27 knockout in zebrafish resulted in craniofacial malformation, spine deformity, and cardiac edema, mirroring typical HFM phenotypes. Abnormalities in somatic cell apoptosis, reduced NCC proliferation in pharyngeal arches, and chondrocyte differentiation issues were observed in cdc27\u2212\/\u2212 mutants. cdc27 mRNA injections and cdkn1a or tp53 knockout significantly rescued pharyngeal arch cartilage dysplasia, while sox9a mRNA administration partially restored the defective phenotypes. Our findings suggest a functional link between CDC27 and HFM, primarily through the inhibition of CNCC proliferation and disruption of pharyngeal chondrocyte differentiation.<\/jats:p>","DOI":"10.3390\/ijms25094707","type":"journal-article","created":{"date-parts":[[2024,4,26]],"date-time":"2024-04-26T05:13:41Z","timestamp":1714108421000},"page":"4707","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Functional and Genetic Analyses Unveil the Implication of CDC27 in Hemifacial Microsomia"],"prefix":"10.3390","volume":"25","author":[{"given":"Wenjie","family":"Song","sequence":"first","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Xia","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Fan","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaowei","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1111\/ocr.12592","article-title":"Craniofacial microsomia\u2014More than a structural malformation","volume":"26","author":"Suominen","year":"2023","journal-title":"Orthod. Craniofac. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/j.ejmg.2005.04.015","article-title":"Oculo-auriculo-vertebral spectrum (OAVS): Clinical evaluation and severity scoring of 53 patients and proposal for a new classification","volume":"48","author":"Tasse","year":"2005","journal-title":"Eur. J. Med. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1177\/0022034518795609","article-title":"Etiology and Pathogenesis of Hemifacial Microsomia","volume":"97","author":"Chen","year":"2018","journal-title":"J. Dent. Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Wang, Y., Ping, L., Luan, X., Chen, Y., Fan, X., Li, L., Liu, Y., Wang, P., Zhang, S., and Zhang, B. (2020). A Mutation in VWA1, Encoding von Willebrand Factor A Domain-Containing Protein 1, Is Associated with Hemifacial Microsomia. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.571004"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"616329","DOI":"10.3389\/fgene.2021.616329","article-title":"ITPR1 Mutation Contributes to Hemifacial Microsomia Spectrum","volume":"12","author":"Liu","year":"2021","journal-title":"Front. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4680","DOI":"10.1038\/s41467-021-24852-9","article-title":"Haploinsufficiency of SF3B2 causes craniofacial microsomia","volume":"12","author":"Timberlake","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1242\/dev.120.3.483","article-title":"Segment and cell type lineage restrictions during pharyngeal arch development in the zebrafish embryo","volume":"120","author":"Schilling","year":"1994","journal-title":"Development"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1006\/dbio.1998.9016","article-title":"The shaping of pharyngeal cartilages during early development of the zebrafish","volume":"203","author":"Kimmel","year":"1998","journal-title":"Dev. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1002\/wdev.147","article-title":"Development and evolution of the pharyngeal apparatus","volume":"3","author":"Frisdal","year":"2014","journal-title":"Wiley Interdiscip. Rev. Dev. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1733","DOI":"10.1097\/SCS.0b013e318254351c","article-title":"Microtia and congenital aural atresia","volume":"23","author":"Genc","year":"2012","journal-title":"J. Craniofac. Surg."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"272","DOI":"10.7150\/ijbs.2929","article-title":"TGF-\u03b2 and BMP signaling in osteoblast differentiation and bone formation","volume":"8","author":"Chen","year":"2012","journal-title":"Int. J. Biol. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"13467","DOI":"10.1074\/jbc.M112.431775","article-title":"TGF-\u03b2-activated kinase 1 (Tak1) mediates agonist-induced Smad activation and linker region phosphorylation in embryonic craniofacial neural crest-derived cells","volume":"288","author":"Yumoto","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1186\/s12935-021-01860-9","article-title":"The importance of CDC27 in cancer: Molecular pathology and clinical aspects","volume":"21","author":"Keramatipour","year":"2021","journal-title":"Cancer Cell Int."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"9210","DOI":"10.1073\/pnas.0913813107","article-title":"C\/EBP\u03b4 targets cyclin D1 for proteasome-mediated degradation via induction of CDC27\/APC3 expression","volume":"107","author":"Pawar","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2626","DOI":"10.7150\/jca.19381","article-title":"CDC27 Induces Metastasis and Invasion in Colorectal Cancer via the Promotion of Epithelial-To-Mesenchymal Transition","volume":"8","author":"Qiu","year":"2017","journal-title":"J. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"10041","DOI":"10.1074\/jbc.M110.205518","article-title":"Phosphorylation of the anaphase-promoting complex\/Cdc27 is involved in TGF-beta signaling","volume":"286","author":"Zhang","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1016\/S0092-8674(03)00432-X","article-title":"Mechanisms of TGF-beta signaling from cell membrane to the nucleus","volume":"113","author":"Shi","year":"2003","journal-title":"Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"938","DOI":"10.1136\/jmedgenet-2021-107695","article-title":"Loeys-Dietz and Shprintzen-Goldberg syndromes: Analysis of TGF-\u03b2-opathies with craniofacial manifestations using an innovative multimodality method","volume":"59","author":"Almpani","year":"2021","journal-title":"J. Med. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1177\/0022034519851786","article-title":"Regulatory Mechanisms of Soft Palate Development and Malformations","volume":"98","author":"Li","year":"2019","journal-title":"J. Dent. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"5065","DOI":"10.1242\/dev.129.21.5065","article-title":"A zebrafish sox9 gene required for cartilage morphogenesis","volume":"129","author":"Yan","year":"2002","journal-title":"Development"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1111\/joa.12821","article-title":"Pbx loss in cranial neural crest, unlike in epithelium, results in cleft palate only and a broader midface","volume":"233","author":"Welsh","year":"2018","journal-title":"J. Anat."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"dmm040279","DOI":"10.1242\/dmm.040279","article-title":"Perturbed development of cranial neural crest cells in association with reduced sonic hedgehog signaling underlies the pathogenesis of retinoic-acid-induced cleft palate","volume":"12","author":"Wang","year":"2019","journal-title":"Dis. Models Mech."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e1633","DOI":"10.1097\/GOX.0000000000001633","article-title":"Genetic Requirement of talin1 for Proliferation of Cranial Neural Crest Cells during Palate Development","volume":"6","author":"Ishii","year":"2018","journal-title":"Plast. Reconstr. Surg. Glob. Open"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Tsai, Y.Y., Su, C.H., and Tarn, W.Y. (2021). p53 Activation in Genetic Disorders: Different Routes to the Same Destination. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22179307"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1038\/nrc2657","article-title":"p21 in cancer: Intricate networks and multiple activities","volume":"9","author":"Abbas","year":"2009","journal-title":"Nat. Rev. Cancer"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e2074","DOI":"10.1038\/cddis.2015.402","article-title":"Downregulation of CDC27 inhibits the proliferation of colorectal cancer cells via the accumulation of p21Cip1\/Waf1","volume":"7","author":"Qiu","year":"2016","journal-title":"Cell Death Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/gim.2015.29","article-title":"Treacher Collins syndrome: A clinical and molecular study based on a large series of patients","volume":"18","author":"Vincent","year":"2016","journal-title":"Genet. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1038\/nm1725","article-title":"Prevention of the neurocristopathy Treacher Collins syndrome through inhibition of p53 function","volume":"14","author":"Jones","year":"2008","journal-title":"Nat. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1038\/ng.724","article-title":"Mutations in genes encoding subunits of RNA polymerases I and III cause Treacher Collins syndrome","volume":"43","author":"Dauwerse","year":"2011","journal-title":"Nat. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1016\/j.bbadis.2016.03.005","article-title":"Pathogenesis of POLR1C-dependent Type 3 Treacher Collins Syndrome revealed by a zebrafish model","volume":"1862","author":"Lau","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1387\/ijdb.170051gc","article-title":"Contribution of cranial neural crest cells to mouse skull development","volume":"61","author":"Wu","year":"2017","journal-title":"Int. J. Dev. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1002\/aja.1002030302","article-title":"Stages of embryonic development of the zebrafish","volume":"203","author":"Kimmel","year":"1995","journal-title":"Dev. Dyn."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.devcel.2018.06.003","article-title":"A Rapid Method for Directed Gene Knockout for Screening in G0 Zebrafish","volume":"46","author":"Wu","year":"2018","journal-title":"Dev. Cell"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Li, L., Mao, A., Wang, P., Ning, G., Cao, Y., and Wang, Q. (2018). Endodermal pouch-expressed dmrt2b is important for pharyngeal cartilage formation. Biol. Open, 7.","DOI":"10.1242\/bio.035444"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Li, X., Ge, G., Song, G., Li, Q., and Cui, Z. (2023). Effects of Nutritionally Induced Obesity on Metabolic Pathways of Zebrafish. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24031850"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Ge, G., Long, Y., Song, G., Li, Q., Cui, Z., and Yan, H. (2022). Transcriptomic Profiling Revealed Signaling Pathways Associated with the Spawning of Female Zebrafish under Cold Stress. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23147494"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1093\/bioinformatics\/btt656","article-title":"featureCounts: An efficient general purpose program for assigning sequence reads to genomic features","volume":"30","author":"Liao","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Love, M.I., Huber, W., and Anders, S. (2014). Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol., 15.","DOI":"10.1186\/s13059-014-0550-8"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"W317","DOI":"10.1093\/nar\/gkab447","article-title":"KOBAS-i: Intelligent prioritization and exploratory visualization of biological functions for gene enrichment analysis","volume":"49","author":"Bu","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Supek, F., Bo\u0161njak, M., \u0160kunca, N., and \u0160muc, T. (2011). REVIGO summarizes and visualizes long lists of gene ontology terms. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0021800"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Hellemans, J., Mortier, G., De Paepe, A., Speleman, F., and Vandesompele, J. (2007). qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol., 8.","DOI":"10.1186\/gb-2007-8-2-r19"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/9\/4707\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:33:30Z","timestamp":1760106810000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/9\/4707"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,4,26]]},"references-count":41,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["ijms25094707"],"URL":"https:\/\/doi.org\/10.3390\/ijms25094707","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,4,26]]}}}