Multi-scale modeling of inherited pediatric cardiomyopathies

项目摘要 该提案的目标是利用基因组编辑和基因组测序技术， 患者来源的iPSC，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。我们重点关注三种单基因心脏病，Barth综合征（BTHS），cate- 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 称为致心肌病性右心室心肌病/发育不良）。这些疾病代表了主要的 遗传性心脏病的类别，即心律失常（CPVT; ACM）和收缩障碍（BTHS; ACM）。没有针对这些疾病的靶向治疗，目前的管理选择也远远不够。 理想，导致悲惨的死亡或心脏移植。我们对这些疾病的研究将突破 体外疾病模型的四个主要方面：（1）通过完善体外系统，以更好地反映生理 （2）通过客观评估诱导多能干细胞衍生的能力， 心肌细胞（iPSC-CM）模型，以捕获患者之间的个体间差异;（3）通过识别新的 通过改进的机制理解或无偏见的筛选进行治疗;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反应是 与哺乳动物模型或患者的反应相比。在UG 3阶段，我们将开发生理学 这三种单基因心脏病的检测系统（目的1）。这些分析系统将从细胞 对三维工程心室，提供了一系列必要的系统，以解决 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UH 3 阶段，我们将使用2D组织和3D心室，通过屏幕和 机制研究（目标2）。我们将使用3D心室执行“培养皿中的临床试验”（目标3）， 确定基于iPSC的模型捕获个体间差异的程度，并测量 一组患者微生理学模型的治疗反应。我们的垂直整合， 多学科方法将汇集心脏生物学家、生物工程师、生物信息学家和临床医生 以推进体外心脏病建模的最新技术。其影响将远远超出 通过改进心脏病模型直接研究三种罕见疾病，并提供有用性数据 用于捕获个体患者表型的iPSC-CM。正常人体器官体外模型的建立 通过提高药物安全性测试的精度和速度，还将大大加快药物开发。 我们的成果包括iPSC-CM分化的进展;用于检测iPSC-CM的新型生物工程系统 生理特性;对三种代表性心脏病发病机制的新见解;以及 用于疾病治疗的治疗靶点和先导化合物的鉴定。

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.