Human Induced Pluripotent Stem Cells for Cardiovascular Disease Modeling

项目总结 在家族性心肌病中，由LMNA突变引起的心脏椎板病占 约占所有病例的6%。与其他有左室扩张的扩张型心肌病患者相比， 心脑膜病患者表现出严重的临床病程、传导异常和高心率。 失败了。LMNA DCM具有复杂的病理生理机制，已假设这种形式的心脏 疾病是由于心肌细胞(CM)功能缺陷以及非CM人群中的缺陷，如 作为内皮细胞(ECs)。然而，LMNA心胺病的详细分子机制仍然存在。 由于患者来源的生物样本有限和缺乏适当的疾病模型，这一点难以捉摸。要克服 问题是，在这次R01拨款续期申请中，我们将产生人类诱导的多能干细胞- LMNA DCM患者来源的心肌细胞(IPSC-CMS)以及匹配的家系对照和健康人 不相关的控件。为了阐明详细的分子机制，我们将进行结构， 电生理、发育、转录组和机制分析也使用特定于患者 为基因组编辑的等基因IPSC-CMS和IPSC-ECs。重要的是，我们将进行药物筛选研究 靶向调节失调的信号通路。为了确认所识别的靶向化合物的有益作用， 然后我们将对等基因LMNA、iPSC-CM和IPSC-ECs进行转录和功能分析 高吞吐量平台。总的来说，这些研究将揭示LMNA的机械性见解 心脏病，DCM的一个主要原因，这可能有助于确定新的候选治疗方法，可以 针对CMS和ECs这两个关键细胞群体。

PROJECT SUMMARY Among the familial cardiomyopathies, cardiac laminopathy caused by LMNA mutations accounts for approximately 6% of all cases. Compared to other DCM patients with left ventricular dilation, LMNA cardiolaminopathy patients exhibit a severe clinical course, conduction abnormalities, and a high rate of heart failure. LMNA DCM has a complex pathophysiology and it has been hypothesized that this form of cardiac disease is due to defects in function of cardiomyocytes (CMs) as well as defects in non-CM populations such as endothelial cells (ECs). However, the detailed molecular mechanisms of LMNA cardiolaminopathy remain elusive due to limited patient-derived bio-specimens and lack of appropriate disease models. To overcome the problem, in this R01 grant renewal application, we will generate human induced pluripotent stem cell- derived cardiomyocytes (iPSC-CMs) from LMNA DCM patients as well as matched family controls and healthy unrelated controls. To clarify the detailed molecular mechanisms, we will conduct structural, electrophysiological, developmental, transcriptome, and mechanistic analyses using patient-specific as well as genome-edited isogenic iPSC-CMs and iPSC-ECs. Importantly, we will perform drug screening studies targeting dysregulated signaling pathways. To confirm the beneficial action of identified targeting compounds, we will then conduct transcriptomic and functional analysis of isogenic LMNA iPSC-CM and iPSC-ECs using high throughput platforms. Collectively, these studies will uncover mechanistic insights of LMNA cardiolaminopathy, a major cause of DCM, which may help identify novel therapeutic candidates that can target CMs and ECs, two key cell populations.