Allele-specific RNA interference rescues the long-QT syndrome phenotype in human-induced pluripotency stem cell cardiomyocytes.
Allele-specific RNA interference rescues the long-QT syndrome phenotype in human-induced pluripotency stem cell cardiomyocytes.
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DOI：
10.1093/eurheartj/eht067
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发表时间：
2014-04
影响因子：
39.3
通讯作者：
Denning C
中科院分区：
文献类型：
--
作者：
Matsa E;Dixon JE;Medway C;Georgiou O;Patel MJ;Morgan K;Kemp PJ;Staniforth A;Mellor I;Denning C
长QT综合征(LQTS)是一种常染色体显性遗传性先天性疾病，与1：1000突变频率、心脏骤停和猝死有关。我们试图使用人类诱导多能干细胞(HiPSCs)来源的心肌细胞作为体外模型来开发和评估基于基因的治疗LQTS的方法。我们在IKR离子通道孔中产生了携带KCNH2 c.G1681A突变的LQTS-2型(LQT2)HiPSC心肌细胞，该突变导致糖基化和通道转运到细胞表面的功能受损。针对突变的KCNH2基因的等位基因特异性RNA干扰(RNAi)导致基因被敲除，而野生型的基因不受影响。经突变特异性siRNAs处理的患者来源的LQT2-hiPSC心肌细胞的电生理分析显示，动作电位时程(APDS)和K+电流正常化，自发性心律失常和药物诱发的心律失常(表现为早-后去极化)同时被挽救。这些发现提供了体外证据，表明等位基因特异性RNAi可以挽救LQTS心肌细胞的病变表型。这是治疗许多常染色体显性遗传阴性疾病的潜在的新途径，包括心脏疾病。
Long-QT syndromes (LQTS) are mostly autosomal-dominant congenital disorders associated with a 1:1000 mutation frequency, cardiac arrest, and sudden death. We sought to use cardiomyocytes derived from human-induced pluripotency stem cells (hiPSCs) as an in vitro model to develop and evaluate gene-based therapeutics for the treatment of LQTS. We produced LQTS-type 2 (LQT2) hiPSC cardiomyocytes carrying a KCNH2 c.G1681A mutation in a IKr ion-channel pore, which caused impaired glycosylation and channel transport to cell surface. Allele-specific RNA interference (RNAi) directed towards the mutated KCNH2 mRNA caused knockdown, while leaving the wild-type mRNA unaffected. Electrophysiological analysis of patient-derived LQT2 hiPSC cardiomyocytes treated with mutation-specific siRNAs showed normalized action potential durations (APDs) and K+ currents with the concurrent rescue of spontaneous and drug-induced arrhythmias (presented as early-afterdepolarizations). These findings provide in vitro evidence that allele-specific RNAi can rescue diseased phenotype in LQTS cardiomyocytes. This is a potentially novel route for the treatment of many autosomal-dominant-negative disorders, including those of the heart.
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