Adeno-associated virus-mediated CASQ2 delivery rescues phenotypic alterations in a patient-specific model of recessive catecholaminergic polymorphic ventricular tachycardia.
Adeno-associated virus-mediated CASQ2 delivery rescues phenotypic alterations in a patient-specific model of recessive catecholaminergic polymorphic ventricular tachycardia.
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DOI：
10.1038/cddis.2016.304
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发表时间：
2016-10-06
影响因子：
9
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中科院分区：
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儿茶酚胺能多态性室性心动过速2型（CPVT 2）是由钙螯合蛋白2（CASQ 2）基因突变引起的高度致死性隐性遗传性心脏病。我们以前已经证明，野生型（WT）CASQ 2基因的病毒转移，防止CPVT 2的发展，在遗传诱导的疾病纯合子携带者的R33 Q突变的小鼠模型。在本研究中，我们研究了病毒介导的基因治疗在从携带纯合CASQ 2-G112+5X突变的患者获得的诱导多能干细胞（iPSC）分化的心肌细胞（CM）中的功效。为此，我们用编码人CASQ 2基因的腺相关病毒载体血清型9（AAV 9）（AAV 9-hCASQ 2）感染细胞。人WT CASQ 2基因的施用能够且足以恢复钙螯合蛋白-2蛋白的生理表达并挽救患者特异性iPSC衍生的CM的功能缺陷。事实上，病毒基因转移后，我们观察到一个显着的百分比下降的延迟后去极化（DAD）的发病CM后肾上腺素能刺激，钙瞬变幅度重新建立和钙火花的密度和持续时间正常化。因此，我们在人类心脏特异性模型系统中证明了AAV 9介导的CPVT 2基因替代疗法的功效，支持了测试的基因疗法在具有不同人类CPVT突变的模型中具有治愈性的观点。
Catecholaminergic Polymorphic Ventricular Tachycardia type 2 (CPVT2) is a highly lethal recessive arrhythmogenic disease caused by mutations in the calsequestrin-2 (CASQ2) gene. We have previously demonstrated that viral transfer of the wild-type (WT) CASQ2 gene prevents the development of CPVT2 in a genetically induced mouse model of the disease homozygous carrier of the R33Q mutation. In the present study, we investigated the efficacy of the virally mediated gene therapy in cardiomyocytes (CMs) differentiated from induced pluripotent stem cells (iPSCs) obtained from a patient carrying the homozygous CASQ2-G112+5X mutation. To this end, we infected cells with an Adeno-Associated Viral vector serotype 9 (AAV9) encoding the human CASQ2 gene (AAV9-hCASQ2). Administration of the human WT CASQ2 gene was capable and sufficient to restore the physiological expression of calsequestrin-2 protein and to rescue functional defects of the patient-specific iPSC-derived CMs. Indeed, after viral gene transfer, we observed a remarkable decrease in the percentage of delayed afterdepolarizations (DADs) developed by the diseased CMs upon adrenergic stimulation, the calcium transient amplitude was re-established and the density and duration of calcium sparks were normalized. We therefore demonstrate the efficacy of the AAV9-mediated gene replacement therapy for CPVT2 in a human cardiac-specific model system, supporting the view that the gene-therapy tested is curative in models with different human mutations of CPVT.
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影响因子：
5.3
作者：
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通讯作者：
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影响因子：
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作者：
通讯作者：
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