先天性房间隔缺损致病基因KLF13新突变的识别与功能分析  

Identification and functional assay of a new KLF13 mutation underpinning congenital atrial

作　　者：陈春英 刘兴元[1] 杨奕清 CHEN Chunying;LIU Xingyuan;YANG Yiqing(Department of Pediatrics,Tongji University School of Medicine,Shanghai 200065;Department of Cardiology,Cardiovascular Research Laboratory,and Central Laboratory,Shanghai Fifth People′s Hospital,Fudan University,Shanghai 200240,China)

机构地区：[1]同济大学医学院附属同济医院儿科,上海200065 [2]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,上海200240

出　　处：《国际心血管病杂志》2023年第2期108-112,共5页International Journal of Cardiovascular Disease

基　　金：上海市自然科学基金(16ZR1432500)。

摘　　要：目的:探索房间隔缺损致病基因KLF13新突变。方法:对175例先天性房间隔缺损患儿和217名健康者的KLF13基因进行测序分析以发现新的致病突变。克隆KLF13基因,构建野生型KLF13表达质粒KLF13-pcDNA3.1,通过定位诱变获得突变型KLF13-pcDNA3.1,转染NIH3T3细胞,通过报告基因分析研究突变体的功能特性。结果:在1例散发性先天性房间隔缺损患儿发现KLF13基因新突变,即NM_015995.4:c.85G>T;p.(Glu29*)突变。该突变不存在于217名健康者。生化分析表明突变型KLF13对靶基因ACTC1的转录激活功能丧失。结论:发现KLF13基因功能丧失性新突变可导致先天性房间隔缺损,这对先天性房间隔缺损的个体化医学防治有潜在的临床意义。Objective:To investigate a novel mutation in the KLF13 gene predisposing to congenital atrial septal defect.Methods:Sequencing analysis of KLF13 was performed in 175 children affected with congenital atrial septal defect and 217 healthy subjects for a new mutation underlying atrial septal defect.The KLF13 gene was cloned and its wild-type expression plasmid KLF13-pcDNA3.1 was constructed.The mutant-type KLF13-pcDNA3.1 plasmid was created via site-targeted mutagenesis.NIH3T3 cells were transiently transfected with various expression plasmids,and the functional characteristics of the mutant-type KLF13 were analyzed by reporter assay.Results:A new KLF13 mutation,NM_015995.4:c.85G>T;p.(Glu29*),was discovered in a child with sporadic congenitalatrial septal defect,which was absent from the 217 healthy individuals.Biological analysis revealed that the Glu29*-mutant KLF13 had no transcriptional activation on its target gene ACTC1.Conclusion:A new loss-of-function KLF13 mutation is identified to cause congenital atrial septal defect,which suggests potential implications for the personalized medical prevention and treatment of congenital atrial septal defect.

关 键 词：房间隔缺损 分子遗传学 转录调节 KLF13基因 报告基因分析 

分 类 号：R725.4[医药卫生—儿科]