Guo Changlong, Wang Qidi, Wang Yuting, Yang Liping, Luo Haiyan, Cao Xiao Fang, An Lisha, Qiu Yue, Du Meng, Ma Xu, Li Hui, Lu Cailing
Department of Genetics, National Research Institute for Family Planning, Haidian, Beijing 100081, P.R. China.
Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
Mol Med Rep. 2017 May;15(5):3193-3197. doi: 10.3892/mmr.2017.6410. Epub 2017 Mar 30.
Genetic variation in specific transcription factors during heart formation may lead to congenital heart disease (CHD) or even miscarriage. The aim of the present study was to identify CHD‑associated genes using next generation sequencing (NGS). The whole exome DNA sequence was obtained from a stillborn fetus diagnosed with tricuspid atresia and complete transposition of the great arteries using high‑throughput sequencing methods. Subsequently, genetic variants of CHD‑associated genes were selected and verified in 215 non‑syndromic CHD patients and 249 healthy control subjects using polymerase chain reaction combined with Sanger sequencing. Genetic variants of previously reported CHD‑inducing genes, such as cysteine rich with EGF like domains 1 and cbp/p300‑interacting transactivator with Glu/Asp rich carboxy‑terminal domain 2, were discovered through the NGS analysis. In addition, a novel non‑synonymous mutation of the iroquois homeobox 1 (IRX1) gene (p.Gln240Glu) was identified. A total of three non‑synonymous mutations (p.Gln240Glu, p.Ser298Asn and p.Ala381Glu) of the IRX1 gene were verified in 215 non‑syndromic CHD patients, but not in 249 healthy volunteers. The results demonstrated that NGS is a powerful tool to study the etiology of CHD. In addition, the results suggest that genetic variants of the IRX1 gene may contribute to the pathogenesis of CHD.
心脏形成过程中特定转录因子的基因变异可能导致先天性心脏病（CHD）甚至流产。本研究的目的是使用下一代测序（NGS）来鉴定与CHD相关的基因。通过高通量测序方法，从一名被诊断为三尖瓣闭锁和大动脉完全转位的死产胎儿中获得了全外显子DNA序列。随后，使用聚合酶链反应结合桑格测序法，在215例非综合征性CHD患者和249名健康对照受试者中选择并验证了与CHD相关基因的遗传变异。通过NGS分析发现了先前报道的CHD诱导基因的遗传变异，如富含EGF样结构域的半胱氨酸1和富含Glu/Asp羧基末端结构域2的cbp/p300相互作用反式激活因子。此外，还鉴定了易洛魁同源框1（IRX1）基因的一个新的非同义突变（p.Gln240Glu）。在215例非综合征性CHD患者中验证了IRX1基因总共三个非同义突变（p.Gln240Glu、p.Ser298Asn和p.Ala381Glu），但在249名健康志愿者中未验证到。结果表明，NGS是研究CHD病因的有力工具。此外，结果表明IRX1基因的遗传变异可能与CHD的发病机制有关。
