Engineered Human Heart Slice for Testing Drug-Induced Arrhythmia

用于测试药物引起的心律失常的工程人体心脏切片

• 批准号: 10250777
• 负责人: LESLIE TUNG
• 金额: $ 1.18万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10250777
• 关键词: Arrhythmia; Arrhythmogenic Right Ventricular Dysplasia; Behavior; CD36 gene; Cardiac; Cardiac Myocytes; Cell Line; Cells; Confounding Factors (Epidemiology); Development; Disease; Disease model; Drug Modelings; Engineering; Entropy; Genetic; Heart; Heart Diseases; Human; Human Engineering; In Vitro; Measures; Metabolic; Metabolic Marker; Methods; Modeling; Nutrient; Pharmaceutical Preparations; Pharmacology; Procedures; Process; Protein Isoforms; Sarcomeres; Slice; Source; Structure; Testing; Time; Tissues; Troponin; Work; base; drug testing; human disease; human model; improved; in vitro Model; induced pluripotent stem cell; insight; novel; parent grant; response; stem cells; therapeutic development; transcriptome sequencing; transcriptomics

The parent grant, “Engineered Human Heart Slice for Testing Drug-Induced Arrhythmia”, aims to develop a pharmacologic testing platform for in vitro prediction of drug-induced cardiac arrhythmia using engineered heart slices (EHS) embedded with human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Aim 1 focuses on improving methods of quantifying and controlling the maturity of hiPSC-CMs, which is key to the implementation and interpretation of these models. The work proposed here will supplement Aim 1 of the parent grant by expanding beyond healthy wildtype hiPSC-CMs to understand maturity in hiPSCs harboring genetic cardiac disorders, specifically arrhythmogenic right ventricular cardiomyopathy (ARVC). The first aim of this work is to establish the maturation trajectory of diseased hiPSC-CMs in comparison to wildtype cells. This will be accomplished by assessing, at several time points, structural features (i.e. sarcomere organization, troponin isoform expression), CD36 positivity (marker of metabolic maturity), and transcriptomic entropy scoring (a novel RNA-sequencing based method). The second aim is to evaluate the effects of targeted metabolic maturation on diseased vs healthy hiPSC-CMs. We will measure maturity in cells treated with standard differentiation procedures and media against cells supplemented with a novel nutrient formula shown to boost maturity in wildtype hiPSC-CMs. By providing insight into the maturational differences of healthy and disease cells and potentially allowing us to account for that confounding variable, this work will facilitate the creation of robust disease models for drug response testing and therapeutic development.

母基金“用于测试药物引起的心律失常的工程人体心脏切片”旨在 开发体外预测药物诱发心脏病的药理学测试平台 使用嵌入人类诱导多能干细胞的工程心脏切片（EHS）治疗心律失常 细胞源性心肌细胞（hiPSC-CM）。目标 1 侧重于改进量化方法 控制 hiPSC-CM 的成熟度，这是实施和解释的关键 这些模型。这里提出的工作将通过扩大家长补助金的目标 1 超越健康野生型 hiPSC-CM 来了解携带心脏遗传基因的 hiPSC 的成熟度 疾病，特别是致心律失常性右心室心肌病（ARVC）。第一个目标是 这项工作的目的是建立患病 hiPSC-CM 的成熟轨迹，并与 野生型细胞。这将通过在几个时间点评估结构特征来完成 （即肌节组织、肌钙蛋白亚型表达）、CD36 阳性（代谢标志物） 成熟度）和转录组熵评分（一种基于 RNA 测序的新型方法）。这 第二个目标是评估目标代谢成熟对患病与健康的影响 hiPSC-CM。我们将测量经过标准分化程序处理的细胞的成熟度 以及补充有新型营养配方的细胞培养基，该配方可促进细胞成熟 野生型 hiPSC-CM。通过深入了解健康和疾病的成熟差异 细胞并可能使我们能够解释该混杂变量，这项工作将有助于 创建用于药物反应测试和治疗开发的强大疾病模型。