Gene Therapy for Catecholaminergic Polymorphic Ventricular Tachycardia by Inhibition of Ca2+/Calmodulin-Dependent Kinase II
Gene Therapy for Catecholaminergic Polymorphic Ventricular Tachycardia by Inhibition of Ca2+/Calmodulin-Dependent Kinase II
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DOI：
10.1161/circulationaha.118.038514
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发表时间：
2019-07-30
期刊：
影响因子：
37.8
通讯作者：
Pu, William T.
中科院分区：
文献类型：
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作者：
Bezzerides, Vassilios J.;Caballero, Ana;Pu, William T.
背景：儿茶酚胺能多形性室性心动过速（CPVT）是一种遗传性心律失常，以肾上腺素能引发的心律失常为特征，目前的护理标准治疗不足。Ca2+/钙调素依赖性蛋白激酶II （CaMKII）是一种肾上腺素激活的激酶，在心脏病模型中有助于心律失常的发生，是CPVT的候选治疗靶点。然而，由于需要在心肌细胞中选择性抑制CaMKII， CaMKII抑制的翻译受到限制。在这里，我们验证了CaMKII抑制与心肌细胞靶向基因治疗策略可以抑制CPVT小鼠模型心律失常的假设。方法：我们开发了AAV9-GFP-AIP，这是一种腺相关病毒载体，其中有效的CaMKII抑制肽，autocamide -2相关抑制肽[AIP]与绿色荧光蛋白（GFP）融合，并从心肌细胞选择性启动子中表达。载体是系统递送的。采用有创电生理试验评价成年小鼠心律失常负荷。我们还对来自不同致病突变的CPVT患者的诱导多能干细胞进行了AIP测试，以确定我们提出的治疗方法对人类诱导多能干细胞来源的心肌细胞和不同致病基因型的有效性。结果：AAV9-GFP-AIP在心脏中有较强表达，在包括脑在内的心外组织中无明显表达。对已知CPVT突变（RYR2(R176Q/+)）的新生小鼠给予AAV9-GFP-AIP，可有效抑制β -肾上腺素能刺激或程序性心室起搏引起的室性心律失常，无明显的促心律失常作用。给青春期小鼠血管内注入AAV9-GFP-AIP可诱导约50%的心肌细胞，并能有效抑制CPVT小鼠的心律失常。来自2名不同致病突变的CPVT患者的诱导多能干细胞衍生的心肌细胞显示异常钙释放事件的频率增加，这被AIP的细胞渗透性形式所抑制。结论：这项概念验证性研究表明，aav介导的CaMKII肽抑制剂递送到心脏可有效抑制CPVT小鼠模型中的心律失常。CaMKII抑制还逆转了具有不同致病突变的人CPVT诱导的多能干细胞衍生的心肌细胞模型中的心律失常表型。
Background: Catecholaminergic polymorphic ventricular tachycardia (CPVT), an inherited cardiac arrhythmia characterized by adrenergically triggered arrhythmias, is inadequately treated by current standard of care. Ca2+/calmodulin-dependent protein kinase II (CaMKII), an adrenergically activated kinase that contributes to arrhythmogenesis in heart disease models, is a candidate therapeutic target in CPVT. However, translation of CaMKII inhibition has been limited by the need for selective CaMKII inhibition in cardiomyocytes. Here, we tested the hypothesis that CaMKII inhibition with a cardiomyocyte-targeted gene therapy strategy would suppress arrhythmia in CPVT mouse models. Methods: We developed AAV9-GFP-AIP, an adeno-associated viral vector in which a potent CaMKII inhibitory peptide, autocamtide-2-related inhibitory peptide [AIP], is fused to green fluorescent protein (GFP) and expressed from a cardiomyocyte selective promoter. The vector was delivered systemically. Arrhythmia burden was evaluated with invasive electrophysiology testing in adult mice. AIP was also tested on induced pluripotent stem cells derived from patients with CPVT with different disease-causing mutations to determine the effectiveness of our proposed therapy on human induced pluripotent stem cell-derived cardiomyocytes and different pathogenic genotypes. Results: AAV9-GFP-AIP was robustly expressed in the heart without significant expression in extracardiac tissues, including the brain. Administration of AAV9-GFP-AIP to neonatal mice with a known CPVT mutation (RYR2(R176Q/+)) effectively suppressed ventricular arrhythmias induced by either beta-adrenergic stimulation or programmed ventricular pacing, without significant proarrhythmic effect. Intravascular delivery of AAV9-GFP-AIP to adolescent mice transduced approximate to 50% of cardiomyocytes and was effective in suppressing arrhythmia in CPVT mice. Induced pluripotent stem cell-derived cardiomyocytes derived from 2 different patients with CPVT with different pathogenic mutations demonstrated increased frequency of abnormal calcium release events, which was suppressed by a cell-permeable form of AIP. Conclusions: This proof-of-concept study showed that AAV-mediated delivery of a CaMKII peptide inhibitor to the heart was effective in suppressing arrhythmias in a murine model of CPVT. CaMKII inhibition also reversed the arrhythmia phenotype in human CPVT induced pluripotent stem cell-derived cardiomyocyte models with different pathogenic mutations.