Multi-scale modeling of inherited pediatric cardiomyopathies

项目总结 这项建议的目标是推进人类心脏病的体外建模，使用基因组编辑和 患者派生的ipscs，使用这些模型来获得对疾病发病机制的新见解，并开发 新的治疗策略。我们专注于三种单基因心脏病，巴特综合征(BTHS)，心脏病- 胆碱能多形性室性心动过速(CPVT)和致心律失常性心肌病(ACM；也 称为致心律失常的右室心肌病/发育不良)。这些疾病代表着严重的 遗传性心脏病的类别，即心律失常(CPVT；ACM)和收缩(BTHS； ACM)。目前还没有针对这些疾病的靶向治疗方法，目前的治疗方案还远远没有 理想的，结果是悲惨的死亡或心脏移植。我们对这些疾病的研究将突破极限 体外疾病模型的研究主要有四种方式：(1)通过提炼体外系统来更好地反映生理学 通过客观评价诱导多能干细胞来源的能力 心肌细胞(IPSC-CM)模型，以捕捉患者之间的个体差异；(3)通过识别新的 通过改进机械理解或无偏见的筛查进行治疗；以及(4)通过执行 概念验证“培养皿中的临床试验”，其中工程细胞和组织模型的响应 与哺乳动物模型或患者的反应相比。在UG3阶段，我们将发展生理学 这三种单基因心脏病的检测系统(目标1)。这些检测系统将从细胞中进行缩放 配对到三维工程脑室，提供必要的系统范围，以满足 从高通量筛选到疾病发病机制再到体外“临床试验”的挑战。在UH3 阶段，我们将使用2D组织和3D脑室通过筛选和 机械论研究(目标2)。我们将使用3D脑室进行“盘中临床试验”(AIM 3)，以 确定基于IPSC的模型捕捉个体间差异的程度，并测量 一组患者微生理模型的治疗反应。我们的垂直整合， 多学科方法将把心脏生物学家、生物工程师、生物信息学家和临床医生聚集在一起。 目的：提高体外心脏疾病模型的研究水平。其影响将远远超出 通过改进心脏疾病模型直接研究三种罕见疾病并提供有用数据 用于捕获个体患者表型的IPSC-CMS。人体正常器官体外模型的建立 还将通过提高药物安全测试的精确度和速度，大大加快药物开发。 我们的成果包括IPSC-CM分化方面的进展；用于检测IPSC-CM的新型生物工程系统 生理特性；对三种典型心脏疾病发病机制的新见解；以及 确定疾病治疗的治疗靶点和先导化合物。

Project summary The goal of this proposal is to advance in vitro modeling of human heart disease using genome-edited and patient-derived iPSCs, to use these models to gain new insights into disease pathogenesis, and to develop new therapeutic strategies. We focus on three monogenic cardiac diseases, Barth syndrome (BTHS), cate- cholaminergic polymorphic ventricular tachycardia (CPVT), and arrhythmogenic cardiomyopathy (ACM; also known as arrhythmogenic right ventricular cardiomyopathy/dysplasia). These disorders represent major classes of inherited heart disease, namely disorders of cardiac rhythm (CPVT; ACM) and contraction (BTHS; ACM). No targeted therapies are available for these disorders, and current management options are far from ideal, resulting in tragic deaths or cardiac transplantation. Our studies of these diseases will push the envelope of in vitro disease models in four principle ways: (1) by refining in vitro systems to better reflect the physiology of native myocardium; (2) by objectively evaluating the ability of induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) models to capture inter-individual variation between patients; (3) by identifying novel therapies through either improved mechanistic understanding or unbiased screening; and (4) by performing proof-of-concept “Clinical trials in a dish”, in which responses of engineered cell and tissue models are compared to responses of mammalian models or patients. In the UG3 Phase, we will develop physiological assay systems of these three monogenic cardiac diseases (Aim 1). These assay systems will scale from cell pairs to three dimensional engineered ventricles, providing the range of systems necessary to address challenges spanning high throughput screening to disease pathogenesis to in vitro “clinical trials”. In the UH3 Phase, we will use the 2D tissues and 3D ventricles to discover novel treatments through screens and mechanistic studies (Aim 2). We will use the 3D ventricles to perform “Clinical trials in a dish” (Aim 3), to determine the extent to which iPSC-based models capture inter-individual variation and to measure the therapeutic responses of a panel of patient microphysiological models. Our vertically integrated, multidisciplinary approach will bring together cardiac biologists, bioengineers, bioinformaticians, and clinicians to advance the state of the art for in vitro cardiac disease modeling. The impact will extend well beyond the three rare diseases directly studied by improving cardiac disease models and providing data on the usefulness of iPSC-CMs for capturing individual patient phenotypes. Creation of in vitro models of normal human organs would also greatly expedite drug development, by increasing the precision and speed of drug safety testing. Our deliverables include advances in iPSC-CM differentiation; novel bioengineered systems to assay iPSC-CM physiological properties; new insights into the pathogenesis of three representative cardiac diseases; and identification of therapeutic targets and lead compounds for disease treatment.