Yuan Qiuping, Zhao Min, Tandon Bhavna, Maili Lorena, Liu Xiaoming, Zhang Anqi, Baugh Evan H, Tran Tam, Silva Renato M, Hecht Jacqueline T, Swindell Eric C, Wagner Daniel S, Letra Ariadne
Department of Pediatrics, University of Texas Health Science Center at Houston Medical School, Houston, Texas.
Pediatric Research Center, University of Texas Health Science Center at Houston Medical School, Houston, Texas.
Mol Genet Genomic Med. 2017 Nov;5(6):730-741. doi: 10.1002/mgg3.332. Epub 2017 Sep 14.
Oligodontia is a severe form of tooth agenesis characterized by the absence of six or more permanent teeth. Oligodontia has complex etiology and variations in numerous genes have been suggested as causal for the condition.
We applied whole-exome sequencing (WES) to identify the cause of oligodontia in a 9-year-old girl missing 11 permanent teeth. Protein modeling and functional analysis in zebrafish were also performed to understand the impact of identified variants on the phenotype.
We identified a novel compound heterozygous missense mutation in WNT10A (c.637G>A:p.Gly213Ser and c.1070C>T:p.Thr357Ile) as the likely cause of autosomal recessive oligodontia in the child. Affected residues are located in conserved regions and variants are predicted to be highly deleterious for potentially destabilizing the protein fold and inhibiting normal protein function. Functional studies in zebrafish embryos showed that wnt10a is expressed in the craniofacies at critical time points for tooth development, and that perturbations of wnt10a expression impaired normal tooth development and arrested tooth development at 5 days postfertilization (dpf). Furthermore, mRNA expression levels of additional tooth development genes were directly correlated with wnt10a expression; expression of msx1, dlx2b, eda, and axin2 was decreased upon wnt10a knockdown, and increased upon wnt10a overexpression.
Our results reveal a novel compound heterozygous variant in WNT10A as pathogenic for oligodontia, and demonstrate that perturbations of wnt10a expression in zebrafish may directly and/or indirectly affect tooth development recapitulating the agenesis phenotype observed in humans.
少牙畸形是一种严重的牙齿发育不全形式,其特征为缺失六颗或更多恒牙。少牙畸形病因复杂,众多基因的变异被认为是导致该病症的原因。
我们应用全外显子组测序(WES)来确定一名缺失11颗恒牙的9岁女孩少牙畸形的病因。还进行了斑马鱼中的蛋白质建模和功能分析,以了解所鉴定变异对表型的影响。
我们在WNT10A基因中鉴定出一种新的复合杂合错义突变(c.637G>A:p.Gly213Ser和c.1070C>T:p.Thr357Ile),这可能是该儿童常染色体隐性少牙畸形的病因。受影响的残基位于保守区域,预计这些变异对蛋白质折叠具有高度有害性,可能会破坏蛋白质折叠并抑制正常蛋白质功能。斑马鱼胚胎中的功能研究表明,wnt10a在牙齿发育的关键时间点在颅面部表达,并且wnt10a表达的扰动会损害正常牙齿发育,并在受精后5天(dpf)使牙齿发育停滞。此外,其他牙齿发育基因的mRNA表达水平与wnt10a表达直接相关;wnt10a敲低后,msx1、dlx2b、eda和axin2 的表达降低;wnt10a过表达后,这些基因的表达增加。
我们的结果揭示了WNT10A基因中的一种新的复合杂合变异是少牙畸形的致病原因,并证明斑马鱼中wnt10a表达的扰动可能直接和/或间接影响牙齿发育,重现了在人类中观察到的发育不全表型。