CaMKII inhibition rectifies arrhythmic phenotype in a patient-specific model of catecholaminergic polymorphic ventricular tachycardia.
CaMKII inhibition rectifies arrhythmic phenotype in a patient-specific model of catecholaminergic polymorphic ventricular tachycardia.
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DOI：
10.1038/cddis.2013.369
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发表时间：
2013-10-10
影响因子：
9
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诱导多能干细胞(IPSC)为人类的发育研究、疾病模型和再生医学方法提供了独特的机会。我们研究的目的是创造一种体外“患者特异性细胞系统”，以便于筛选新的治疗分子来治疗儿茶酚胺能多形性室性心动过速(CPVT)，这是一种遗传性心律失常。在这里，我们报道了通过从一名携带心脏兰诺定受体基因(RyR2)杂合突变的CPVT患者产生IPSC并随后将其分化为心肌细胞(CMS)来发展CPVT的心脏模型。全细胞膜片钳和自发性搏动细胞的细胞内电记录显示，静息状态和β肾上腺素能刺激后，CPVT-CMS均存在延迟后除极(DAD)，与患者的心脏表型相似。此外，KN-93(2-[N-(2-hydroxyethyl)]-N-(4methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine)，是一种抑制钙/钙调蛋白依赖性丝氨酸苏氨酸蛋白激酶II(CaMKII)的抗心律失常药物，显著减少了CVPT-CMS中DADS的存在，挽救了儿茶酚胺能应激诱导的心律失常表型。此外，通过快速分辨光学作图对3d节拍星团进行的细胞内钙瞬变测量表明，CPVT星团发生了多个钙瞬变，而在野生型星团中，只检测到了单一的启动。这种不稳定性在异丙肾上腺素存在时加剧，并被KN-93减弱。正如在我们的RyR2基因敲除CPVT小鼠身上看到的那样，KN-93的抗心律失常作用在这些人IPSC来源的心肌细胞中得到证实，支持这一体外系统在药物筛选和临床治疗策略优化中的作用。
Induced pluripotent stem cells (iPSC) offer a unique opportunity for developmental studies, disease modeling and regenerative medicine approaches in humans. The aim of our study was to create an in vitro ‘patient-specific cell-based system' that could facilitate the screening of new therapeutic molecules for the treatment of catecholaminergic polymorphic ventricular tachycardia (CPVT), an inherited form of fatal arrhythmia. Here, we report the development of a cardiac model of CPVT through the generation of iPSC from a CPVT patient carrying a heterozygous mutation in the cardiac ryanodine receptor gene (RyR2) and their subsequent differentiation into cardiomyocytes (CMs). Whole-cell patch-clamp and intracellular electrical recordings of spontaneously beating cells revealed the presence of delayed afterdepolarizations (DADs) in CPVT-CMs, both in resting conditions and after β-adrenergic stimulation, resembling the cardiac phenotype of the patients. Furthermore, treatment with KN-93 (2-[N-(2-hydroxyethyl)]-N-(4methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine), an antiarrhythmic drug that inhibits Ca2+/calmodulin-dependent serine–threonine protein kinase II (CaMKII), drastically reduced the presence of DADs in CVPT-CMs, rescuing the arrhythmic phenotype induced by catecholaminergic stress. In addition, intracellular calcium transient measurements on 3D beating clusters by fast resolution optical mapping showed that CPVT clusters developed multiple calcium transients, whereas in the wild-type clusters, only single initiations were detected. Such instability is aggravated in the presence of isoproterenol and is attenuated by KN-93. As seen in our RyR2 knock-in CPVT mice, the antiarrhythmic effect of KN-93 is confirmed in these human iPSC-derived cardiac cells, supporting the role of this in vitro system for drug screening and optimization of clinical treatment strategies.