Wang Yibei, Ping Lu, Luan Xiaodong, Chen Yushan, Fan Xinmiao, Li Lianyan, Liu Yaping, Wang Pu, Zhang Shuyang, Zhang Bo, Chen Xiaowei
Department of Otolaryngology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Department of Otolaryngology, China-Japan Friendship Hospital, Beijing, China.
Front Cell Dev Biol. 2020 Sep 9;8:571004. doi: 10.3389/fcell.2020.571004. eCollection 2020.
Hemifacial microsomia (HFM) is a type of rare congenital syndrome caused by developmental disorders of the first and second pharyngeal arches that occurs in one out of 5,600 live births. There are significant gaps in our knowledge of the pathogenic genes underlying this syndrome.
Whole exome sequencing (WES) was performed on five patients, one asymptomatic carrier, and two marry-in members of a five-generation pedigree. Structure of WARP (product of ) was predicted using the Phyre2 web portal. hybridization and -knockdown/knockout studies in zebrafish using morpholino and CRISPR/Cas9 techniques were performed. Cartilage staining and immunofluorescence were carried out.
Through WES and a set of filtration, we identified a c.G905A:p.R302Q point mutation in a novel candidate pathogenic gene, . The Phyre2 web portal predicted alterations in secondary and tertiary structures of WARP, indicating changes in its function as well. Predictions of protein-to-protein interactions in five pathways related to craniofacial development revealed possible interactions with four proteins in the FGF pathway. Knockdown/knockout studies of the zebrafish revealed deformities of pharyngeal cartilage. A decrease of the proliferation of cranial neural crest cells (CNCCs) and alteration of the structure of pharyngeal chondrocytes were observed in the morphants as well.
Our data suggest that a mutation in is functionally linked to HFM through suppression of CNCC proliferation and disruption of the organization of pharyngeal chondrocytes.
半侧颜面短小畸形（HFM）是一种由第一和第二咽弓发育障碍引起的罕见先天性综合征，在每5600例活产中出现1例。我们对该综合征潜在致病基因的了解存在重大差距。
对一个五代家系的5名患者、1名无症状携带者和2名嫁入成员进行了全外显子组测序（WES）。使用Phyre2门户网站预测WARP（ 产物）的结构。使用吗啉代和CRISPR/Cas9技术在斑马鱼中进行杂交和 -敲低/敲除研究。进行软骨染色和免疫荧光检测。
通过WES和一系列筛选，我们在一个新的候选致病基因中鉴定出一个c.G905A:p.R302Q点突变。Phyre2门户网站预测WARP的二级和三级结构发生改变，表明其功能也发生了变化。对与颅面发育相关的五条途径中的蛋白质 - 蛋白质相互作用的预测揭示了与FGF途径中的四种蛋白质可能存在相互作用。斑马鱼的敲低/敲除研究显示咽软骨畸形。在吗啉代处理的胚胎中也观察到颅神经嵴细胞（CNCCs）增殖减少和咽软骨细胞结构改变。
我们的数据表明， 中的突变通过抑制CNCC增殖和破坏咽软骨细胞的组织而在功能上与HFM相关。
