Tian Hua, Feng Jifan, Li Jingyuan, Ho Thach-Vu, Yuan Yuan, Liu Yang, Brindopke Frederick, Figueiredo Jane C, Magee William, Sanchez-Lara Pedro A, Chai Yang
Center for Craniofacial Molecular Biology, University of Southern California, Los Angeles, CA 90033, USA.
Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology, Beijing 100081, China.
Hum Mol Genet. 2017 Mar 1;26(5):860-872. doi: 10.1093/hmg/ddx002.
Ciliopathies are pleiotropic human diseases resulting from defects of the primary cilium, and these patients often have cleft lip and palate. IFT88 is required for the assembly and function of the primary cilia, which mediate the activity of key developmental signaling pathways. Through whole exome sequencing of a family of three affected siblings with isolated cleft lip and palate, we discovered that they share a novel missense mutation in IFT88 (c.915G > C, p.E305D), suggesting this gene should be considered a candidate for isolated orofacial clefting. In order to evaluate the function of IFT88 in regulating craniofacial development, we generated Wnt1-Cre;Ift88fl/fl mice to eliminate Ift88 specifically in cranial neural crest (CNC) cells. Wnt1-Cre;Ift88fl/flpups died at birth due to severe craniofacial defects including bilateral cleft lip and palate and tongue agenesis, following the loss of the primary cilia in the CNC-derived palatal mesenchyme. Loss of Ift88 also resulted in a decrease in neural crest cell proliferation during early stages of palatogenesis as well as a downregulation of the Shh signaling pathway in the palatal mesenchyme. Importantly, Osr2KI-Cre;Ift88fl/flmice, in which Ift88 is lost specifically in the palatal mesenchyme, exhibit isolated cleft palate. Taken together, our results demonstrate that IFT88 has a highly conserved function within the primary cilia of the CNC-derived mesenchyme in the lip and palate region in mice and is a strong candidate as an orofacial clefting gene in humans.
纤毛病是由初级纤毛缺陷导致的多效性人类疾病,这些患者常伴有唇腭裂。IFT88是初级纤毛组装和功能所必需的,初级纤毛介导关键发育信号通路的活性。通过对一个有三名患单纯性唇腭裂的患病兄弟姐妹的家庭进行全外显子组测序,我们发现他们在IFT88基因中共享一个新的错义突变(c.915G > C,p.E305D),这表明该基因应被视为单纯性口面部裂隙的一个候选基因。为了评估IFT88在调节颅面发育中的功能,我们构建了Wnt1-Cre;Ift88fl/fl小鼠,以特异性地消除颅神经嵴(CNC)细胞中的Ift88。Wnt1-Cre;Ift88fl/fl幼崽在出生时死亡,原因是严重的颅面缺陷,包括双侧唇腭裂和舌发育不全,这是由于CNC来源的腭间充质中初级纤毛的缺失所致。Ift88的缺失还导致腭发育早期神经嵴细胞增殖减少以及腭间充质中Shh信号通路的下调。重要的是,Osr2KI-Cre;Ift88fl/fl小鼠(其中Ift88在腭间充质中特异性缺失)表现出单纯性腭裂。综上所述,我们的结果表明,IFT88在小鼠唇和腭区域CNC来源的间充质的初级纤毛中具有高度保守的功能,并且是人类口面部裂隙基因的有力候选者。