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“用于测试药物诱导心律失常的工程化人类心脏切片”旨在 建立体外预测药物性心脏损害的药理学试验平台 使用嵌入人诱导多能干细胞的工程化心脏切片（EHS）进行心律失常研究 细胞衍生的心肌细胞（hiPSC-CM）。目标1侧重于改进量化方法 以及控制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 超越健康的野生型hiPSC-CM，以了解携带遗传性心脏病的hiPSC的成熟 在某些实施方案中，所述方法用于治疗疾病，特别是致心律失常性右心室心肌病（ARVC）。的首要目标 这项工作是建立患病hiPSC-CM的成熟轨迹， 野生型细胞这将通过在多个时间点评估结构特征来实现 (i.e.肌节组织，肌钙蛋白亚型表达），CD 36阳性（代谢标志物 成熟度）和转录组熵评分（一种基于RNA测序的新方法）。的 第二个目的是评估靶向代谢成熟对疾病与健康的影响。 hiPSC-CM。我们将测量用标准分化程序处理的细胞的成熟度 和培养基对细胞补充了一种新的营养配方，显示出提高成熟， 野生型hiPSC-CM。通过提供健康和疾病的成熟差异的洞察力 细胞，并可能使我们能够考虑到这一混淆变量，这项工作将有助于 为药物反应测试和治疗开发创建强大的疾病模型。