Genome Editing of Human iPSCs to Study Inherited Hypertrophic Cardiomyopathy

 描述(申请人提供)：家族性肥厚型心肌病(HCM)是一种可遗传的常染色体显性遗传特征，表现为心肌异常增厚和组织学表现。到目前为止，已有大量研究揭示了基因突变与肥厚性心肌病之间的相关性。然而，由于从该疾病患者获得心肌细胞(CMS)的途径有限，潜在的发育病因在很大程度上仍不清楚。近年来，人类诱导多能干细胞(IPSCs)从皮肤或血液中分化为心肌细胞(iPSC-CMS)的研究取得了突破性进展，为疾病模型的研究提供了新的来源。使用这一策略，我们有一个独特的机会来研究肥厚型心肌炎的发育性病因学。这项建议将证明用患者特定的IPSC-CMS和基因组编辑的IPSC-CMS来阐明肌丝疾病的可行性。目标1是从HCM突变患者和匹配的家庭对照以及无关的健康捐赠者中产生IPSCs。目的2是通过对IPSC-CMS进行分子和功能分析来确定疾病的程度。目的3是用基因组编辑技术概括肉瘤肉瘤病的表型。总之，我们相信，这项研究的发现将为我们理解肥厚性心肌病的发展病因学提供新的见解，并为肥厚性心肌病的治疗策略带来潜在的新靶点。

 DESCRIPTION (provided by applicant): Familial hypertrophic cardiomyopathy (HCM) is an inheritable autosomal dominant trait, displaying abnormal thickening of myocardium and histologic manifestations. To date, considerable studies have uncovered the correlation between gene mutations and HCM. However, the underlying developmental etiology remains largely unknown due to the limited access to obtain cardiomyocytes (CMs) from patients with this disease. Recent breakthrough in generation of human induced pluripotent stem cells (iPSCs) from skin or blood and de novo differentiation into cardiomyocytes (iPSC-CMs) offers a novel source for disease modeling. Using this strategy, we have a unique opportunity to study the developmental etiology of HCM. This proposal will demonstrate the feasibility of elucidating myofilament diseases with patient-specific iPSC-CMs and genome-edited iPSC-CMs. Aim 1 is to generate iPSCs from patients with HCM mutations and matched family controls as well as unrelated healthy donors. Aim 2 is to determine the extent of disease by performing molecular and functional analyses of iPSC-CMs. Aim 3 is to recapitulate sarcomeric HCM disease phenotype with genome editing technology. In summary, we believe findings from this study will yield novel insights into our understanding of the developmental etiology of HCM and give rise to potential new targets for therapeutic strategy of HCM.