Modeling Familial Dilated Cardiomyopathy Disease Mechanism Using Human iPS Cells

描述（由申请人提供）：心力衰竭是工业化国家最普遍的发病和死亡原因。家族性扩张型心肌病（DCM）是美国和世界范围内最常见的心力衰竭原因。测序技术的最新进展进一步发现了33种已知的与DCM相关的遗传突变。其中，80%以上可在LMNA、MYH6、MYH7、MYPN、TNNT2、SCN5a、MYBPC3这7个基因中找到。合乎逻辑的下一步是研究基因型与疾病背后的机制基础之间的相关性。然而，由于缺乏适当的疾病模型，这些研究一直受到阻碍。在本提案中，我将从DCM患者中生成人类诱导多能干细胞来源的心肌细胞（hiPSC-CMs）作为疾病模型，并进行详细的机制分析，以确定DCM的功能和分子表型。具体来说，hipsc将来自健康对照和三个具有MYH6、TNNT2和MYBPC3遗传突变的家族队列。这些hiPSC细胞系随后将分化为心肌细胞。接下来，对照和DCM患者分化的hiPSC- CMs将通过检查形态学和肌细胞结构、电生理、细胞力学和基因表达谱进行比较。最后，我将从对照组和患者队列中检查hiPSC-CMs对药物治疗和基因治疗的反应。总之，这些研究应该为提高我们未来进行高通量药物筛选、评估基因和细胞疗法以及评估潜在的DCM新疗法的能力铺平道路。

DESCRIPTION (provided by applicant): Heart failure represents the most prevalent cause of morbidity and mortality in the industrialized countries. Familial dilated cardiomyopathy (DCM) is the most common cause of heart failure in the US and worldwide. Recent advances in sequencing technology have further enabled the discovery of 33 known inherited mutations associated with DCM. Of these, more than 80% can be found in 7 genes, namely LMNA, MYH6, MYH7, MYPN, TNNT2, SCN5a, and MYBPC3. The logical next step is to study the correlation between genotype and the mechanistic basis behind the diseases. However, these studies have been hindered by the lack of appropriate disease models. In this proposal, I will generate human induced pluripotent stem cell- derived cardiomyocytes (hiPSC-CMs) from patients with DCM as disease models and perform detailed and mechanistic analyses to determine the functional and molecular phenotypes of DCM. Specifically, hiPSCs will be derived from healthy control and three family cohorts with inherited mutations in MYH6, TNNT2, and MYBPC3. These hiPSC lines will then be differentiating into cardiomyocytes. Next, the differentiated hiPSC- CMs from control and DCM patients will be compared by examining the morphology and myocyte architecture, electrophysiology, cellular mechanics, and gene expression profile. Finally, I will examine the response of the hiPSC-CMs from control and patient cohorts to pharmacologic treatment and gene therapy. In summary, these studies should pave the way to enhance our ability to perform future high-throughput drug screening, evaluate gene and cell therapies, and assess potential novel therapies of DCM.