Yu Xiaoning, Chen Binbin, Zhang Xin, Shentu Xingchao
Eye Center, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Mol Vis. 2017 Apr 28;23:296-305. eCollection 2017.
To test for the potential presence of novel mutations in the () gene in patients with sporadic keratoconus (KC) from a Han Chinese population.
Fifty-three patients with primary KC, 30 patients with high myopia (HM), and 100 unrelated population-matched healthy controls without any ocular or systemic disorders, all of Han Chinese ethnicity, were recruited. Blood samples were donated, and genomic DNA was isolated from peripheral blood leukocytes. Sequence variations in were initially identified in patients with KC with next-generation sequencing and subsequently confirmed using Sanger sequencing. Sequence variants identified in patients with KC were subsequently screened in 30 patients with HM and 100 healthy control subjects. Other genes that were reported to be related to KC were also screened in the patients with KC who carried the mutations in . The Sorting Intolerant Form Tolerant (SIFT) program was used to predict the effect of amino acid substitution on the ZNF469 protein.
Sixteen sequence variants in the coding regions of were identified in this Chinese KC cohort. After five known single nucleotide polymorphisms (SNPs), one false-positive result, and three mutations that were also detected in the results of the whole-exome sequencing (WES) data performed in 220 Han Chinese individuals without ocular abnormalities were removed, seven novel mutations in (c.2059G>A, c.2137C>A, c.3466G>A, c.3749C>T, c.4300G>A, c.4684G>A, and c.7262G>A) that were predicted to be potentially damaging were identified. The patient with KC with the c.3466G>A mutation was also shown to carry one dedicator of cytokinesis 9 () mutation (c.1940C>T). None of the mutations were detected in the patients with HM or the healthy controls. All of the seven mutations in the patients with KC were heterozygote.
The results suggested for the first time that has a pathogenic role in Chinese patients with KC and have widened the mutation spectrum of KC in the Han Chinese population.
检测来自中国汉族人群的散发性圆锥角膜（KC）患者中（）基因是否存在新的突变。
招募了53例原发性KC患者、30例高度近视（HM）患者和100名无任何眼部或全身疾病的汉族人群匹配健康对照。采集血样，从外周血白细胞中分离基因组DNA。首先通过下一代测序在KC患者中鉴定基因序列变异，随后使用桑格测序进行确认。在30例HM患者和100名健康对照中对KC患者中鉴定出的序列变异进行筛查。对携带该基因突变的KC患者还筛查了其他据报道与KC相关的基因。使用不耐受排序到耐受（SIFT）程序预测氨基酸取代对ZNF469蛋白的影响。
在这个中国KC队列中鉴定出该基因编码区的16个序列变异。去除5个已知单核苷酸多态性（SNP）、1个假阳性结果以及在220名无眼部异常的汉族个体进行的全外显子测序（WES）数据结果中也检测到的3个突变后，鉴定出该基因的7个新突变（c.2059G>A、c.2137C>A、c.3466G>A、c.3749C>T、c.4300G>A、c.4684G>A和c.7262G>A），这些突变预计具有潜在损害性。携带c.3466G>A突变的KC患者还被发现携带一个胞质分裂9（）基因突变（c.1940C>T）。在HM患者或健康对照中未检测到任何突变。KC患者中的所有7个突变均为杂合子。
结果首次表明该基因在中国KC患者中具有致病作用，并拓宽了汉族人群中KC的突变谱。
