一例先天性心脏缺损相关ISL1基因新变异的发现及功能分析  

Discovery and functional analysis of a novel ISL1 variant associated with congenital heart defect

作　　者：董斌斌 刘兴元[2] 王天明 杨奕清 Dong Binbin;Liu Xingyuan;Wang Tianming;Yang Yiqing(Department of Pediatrics,Huashan North Hospital,Fudan University,Shanghai 201907,China;Department of Pediatrics,Tongji Hospital,Tongji University,Shanghai 200065,China;Department of Cardiovascular Research,Shanghai Fifth People’s Hospital,Fudan University,Shanghai 200240,China)

机构地区：[1]复旦大学附属华山医院北院儿科,上海201907 [2]同济大学附属同济医院儿科,上海200065 [3]复旦大学附属上海市第五人民医院心血管研究室,上海200240

出　　处：《中华医学遗传学杂志》2020年第9期972-975,共4页Chinese Journal of Medical Genetics

基　　金：上海市自然科学基金(16ZR1432500)。

摘　　要：目的分析与先天性心脏缺损(congenital heart defect,CHD)相关的ISL1基因变异及其功能特点。方法收集194例CHD患者和232名正常对照的临床资料和外周血样,提取基因组DNA。测序分析全部对象的ISL1基因的编码外显子及侧翼的部分内含子。构建野生型ISL1基因表达质粒ISL1-pcDNA3.1,通过定位诱变获得相应的变异体。应用脂质体将基因表达质粒转染CHO细胞,应用双荧光素酶报告基因分析试剂研究变异型ISL1的功能特性。结果在1例散发性房间隔缺损患者中检测出1种新的杂合性ISL1基因变异c.499C>T(p.Q167X)。功能研究表明变异型ISL1对靶基因MEF2C启动子的转录激活功能丧失。结论发现了一个与CHD相关的ISL1基因新变异,ISL1基因的缺陷可能为部分CHD的分子病因。Objective To analyze variation of ISL1 gene and explore its functional characteristics in relation with congenital heart defect(CHD).Methods Clinical data and peripheral blood samples of 194 CHD patients and 232 healthy controls were collected for the extraction of genomic DNA.The coding exons and flanking intronic regions of the ISL1 gene were sequenced.Expression plasmid for the wild-type ISL1 gene ISL1-pcDNA3.1 was constructed,and the corresponding variants were obtained by site-specific mutagenesis.The gene expression plasmid was transfected into CHO cells with liposome,and the functional characteristics of ISL1 variant were studied by double luciferase reporter gene analysis.Results A novel variant of the ISL1 gene c.499C>T(p.Q167X)was detected in a patient with sporadic CHD.Functional study showed that the variant has lost its transcriptional activation function for the MEF2C promoter.Conclusion A novel variant of the ISL1 gene related to CHD has been identified.The defect of ISL1 gene may underlay the pathogenesis for a proportion of CHD.

关 键 词：先天性心脏缺损 分子遗传学 转录因子 ISL1基因 报告基因分析 

分 类 号：R725.4[医药卫生—儿科]