全外显子组测序筛选先天性心脏病基因突变的研究

Whole-exome sequencing reveals genetic mutations in congenital heart disease

目的用全外显子组学测序技术捕获和筛选新生儿先天性心脏病的基因突变。方法于2014年1月—2018年4月,从兰州市妇幼保健院新生儿科明确诊断为以室缺伴房缺的严重先天性心脏病患儿中选取6例为研究对象,收集患儿与父母家系血样,通过全外显子测序技术捕获突变基因,采用公共数据库dbsnp、HapMap以及VEP和BGI内部数据库进行注释和筛选定位先天性心脏病的致病突变,用Sanger测序验证突变。结果在6个家庭中有3个家庭发现了符合筛选条件的新生基因突变,其中3个基因发生错义突变,分别为FH2域控制蛋白1(FHDC1)(c.1977G>C;p.Gln659His)、溴区包含蛋白4(BRD4)(c.3797A>C;p.Glu1266Ala)和己糖激酶3(HK3)(c.397C>G;p.Leu133Val),一个基因剪切位点发生突变为sprouty相关EVH1域包含蛋白2(SPRED2)(c.373+1G>T)。结论FHDC1(c.1977G>C)、BRD4(c.3797A>C)、HK3(c.397C>G)、SPRED2(c.373+1G>T)可能为先天性心脏病有害的新生突变。

Objective Whole exome sequencing was used to capture and screen genetic mutations in neonatal congenital heart disease.MethodsWe selected 6 children with severe congenital heart disease who had ventricular septal defect and atrial septal defect diagnosed by neonatology specialists from January 2014 to April 2018 in Lanzhou Maternal and Child Health Hospital.Blood samples of 6 children and their parents were collected,and the mutant genes were captured by whole exome sequencing.The dbSNP,HapMap,and internal databases of VEP and BGI were used to annotate and screen the pathogenic mutations of congenital heart disease.The mutations were verified by Sanger sequencing.ResultsAccording to the screening criteria,de novo mutations were found in the three of the six families.Three of them have missense mutations,namely FH2 domain-containing protein 1(FHDC1)(c.1977G>C;p.Gln659His)、bromodomain-containing protein 4(BRD4)(c.3797A>C;p.Glu1266Ala)、hexokinase 3(HK3)(c.397C>G;p.Leu133Val),as well as one gene splice site mutated to sprouty related EVH1 domain-containing protein 2(SPRED2)(c.373+1G>T).ConclusionFHDC1(c.1977G>c),BRD4(c.3797A>c),HK3(c.397C>G),and SPRED2(c.373+1G>T)may be deleterious de novo mutations for congenital heart disease.