孤立性心房颤动致病基因TBX20新突变的发现及功能研究
Identification and functional investigation of a new TBX20 mutation contributing to lone atrial fibrillation
张道良 1李宁 2姜伟峰 3吴绍辉 3仇兴标 3杨奕清4
作者信息
- 1. 518057 中国医学科学院阜外医院深圳医院心律失常中心
- 2. 200062 上海中医药大学附属普陀医院心内科
- 3. 200030 上海交通大学医学院附属胸科医院心内科
- 4. 200240 复旦大学附属上海市第五人民医院心内科、心血管研究室、中心实验室
- 折叠
摘要
目的:寻找孤立性心房颤动(房颤)致病基因TBX20 新突变并研究其功能.方法:测序分析 182 例孤立性房颤患者和 236 名无房颤对照者的TBX20 基因,以发现新的致房颤突变.克隆人TBX20基因,构建野生型TBX20 表达质粒,通过定点诱变制备突变型TBX20表达质粒,借助脂质体将表达质粒转染HeLa细胞,应用双荧光素酶报告基因分析系统研究突变体的功能特性.结果:在 1 例散发性孤立性房颤患者中发现TBX20 基因新突变,即NM_001077653.2:c.706A＞T;p.(Lys236*)突变.该突变不存在于其他孤立性房颤患者和对照者.功能研究显示突变型TBX20 对靶基因KCNH2 的转录激活作用丧失.结论:TBX20基因功能障碍可能是部分房颤患者的分子病因,这对房颤的精准防治有潜在临床意义.
Abstract
Objective:To identify and functionally investigate a new TBX20 mutation underlying lone atrial fibrillation.Methods:Sequencing assay of TBX20 was performed in 182 patients with lone atrial fibrillation and 236 control individuals without atrial fibrillation to identify a new mutation accountable for atrial fibrillation.The human TBX20 gene was cloned with its wild-type expression plasmid constructed.The mutant-type TBX20 expression plasmid was produced through site-directed mutagenesis.Hela cells were transfected using multiple expression plasmids with the help of lipofectamine and the functional characteristics of the mutant-type TBX20 were explored by utilizing a dual-reporter assay system.Results:In one patient suffering from sporadic lone atrial fibrillation,a new TBX20 mutation,NM_001077653.2:c.706A＞T;p.(Lys236*),was found,which was neither observed in the other patients with lone atrial fibrillation nor detected in the control subjects.Further functional investigation revealed that the mutant-type TBX20 lost the transactivation effect on its target gene KCNH2.Conclusion:Dysfunctional TBX20 is likely to be a molecular cause of atrial fibrillation in a minority of cases,implying its potential clinical significance for the precise prophylaxis and therapy of atrial fibrillation.
关键词
心房颤动/分子遗传学/转录因子/TBX20基因/报告基因分析Key words
Atrial fibrillation/Molecular genetics/Transcriptional factor/TBX20/Report gene analysis引用本文复制引用
基金项目
国家自然科学基金(82070331)
广东省引进创新创业团队项目(2019ZT08Y481)
出版年
2024
NETL
NSTL
万方数据