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: 10593346
• 负责人: LESLIE TUNG
• 金额: $ 5.89万
• 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 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10593346
• 关键词: Academia; Adopted; Adoption; Advocate; Affect; Aleurites; Area; Arrhythmia; Bioinformatics; Biological Assay; CD36 gene; Cardiac; Cardiac Electrophysiologic Techniques; Cardiac Myocytes; Cardiac development; Cell Maturation; Cell model; Cell surface; Cells; Cellular Assay; Clinical; Complement; Consensus; Dependence; Development; Developmental Biology; Drug Interactions; Engineering; Event; Exhibits; Experimental Models; Fingerprint; Formulation; Gene Expression Profiling; Genetic Transcription; Giant Cells; Goals; Government; Growth and Development function; Guidelines; Heart; Heterogeneity; Histologic; Human; Human Engineering; In Vitro; Incidence; Industry; Ion Channel; Knowledge; Life; Maintenance; Measurement; Metabolic; Modeling; Myocardium; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Physiological; Positioning Attribute; Regulation; Research; Research Contracts; Rest; Risk; Risk Factors; Safety; Signal Pathway; Slice; Stimulant; System; Tachyarrhythmias; Technology; Testing; Time; Tissue Model; Tissues; Torsades de Pointes; United States Food and Drug Administration; Validation; Variant; Ventricular Tachycardia; Verapamil; base; drug testing; falls; high risk; high throughput screening; improved; in vitro Model; in vivo; induced pluripotent stem cell derived cardiomyocytes; novel; novel therapeutics; pre-clinical; public-private partnership; research and development; response; safety testing

New standards are under development for cardiac safety testing of drugs for risk of arrhythmia, ultimately with the preclinical goal of predicting risk for Torsade de Pointes (TdP). In the U.S. the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative has been advocated by government regulatory agencies, public-private partnerships, industry and academia. One component of the initiative rests on the usage of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as a validation platform of drug responses that can replace heterologous expression systems which dwell on block of a single ion channel (HERG). If adopted by the FDA, CiPA represents a major step forward for drug testing using cells that contain the major ion channels and signaling pathways found in human heart. This application of hiPSC-CMs is an area of active research and development, and numerous contract research organizations and most pharmaceutical companies have founded labs to perform high throughput screening of test compounds using these cells. However, current hiPSC-CM screens tend to be on single or small numbers of cells that exhibit immaturity and heterogeneity. In addition, arrhythmic events are limited to early afterdepolarizations (EADs) or irregular beating. TdP is ultimately a multicellular tissue phenomenon that results from reentrant or multifocal ectopic activity that require a minimum size to be revealed. The goal of this project is to develop an in vitro model suitable for testing pharmacological compounds and assessing risk for tissue-level arrhythmia. Three research labs at Hopkins will contribute their expertise for the project. The tissue platform used to support the hiPSC-CMs will be the Engineered Heart Slice developed in Tung’s cardiac electrophysiology lab. Version 2.0 of the slice with improved physiological function will be implemented using proven and putative maturation stimulants, and then used to test for drug-induced reentrant or multifocal arrhythmia. To this end, metabolic maturation of the cells will be pursued via the expertise of the Boheler lab, which has identified CD36 as a cell surface marker distinguishing cells that are metabolically more mature. A bioinformatics approach developed in Kwon’s cardiac developmental biology lab will be used to follow the maturation trajectory of the cardiomyocytes. The convergence of these technologies will result in an advanced in vitro tissue model that will be used to independently and mechanistically assess the proarrhythmic effects of selected compounds which have been identified clinically as low, intermediate, and high risk, and to determine whether the maturation state of the cardiomyocytes affects their drug responses. This approach represents a new paradigm for cardiac safety testing – one that is a step closer to predicting TdP-like arrhythmic events in the myocardium – and may establish new standards for the utility, validity, and maturation-dependence of hiPSC-CMs as an experimental model to assess arrhythmia risk.

正在制定针对心律失常风险药物的心脏安全测试的新标准，最终 临床前目标是预测尖端扭转型室性心动过速 (TdP) 的风险。在美国，综合体外 致心律失常检测 (CiPA) 倡议得到了政府监管机构、公私部门的倡导 伙伴关系、工业界和学术界。该计划的一个组成部分在于使用人类诱导的 多能干细胞衍生的心肌细胞（hiPSC-CM）作为药物反应的验证平台，可以 取代驻留在单离子通道 (HERG) 阻断上的异源表达系统。如果被采纳 FDA、CiPA 代表着使用含有主要离子通道和 人类心脏中发现的信号通路。 hiPSC-CM 的这一应用是一个活跃的研究领域 开发，许多合同研究组织和大多数制药公司都成立了 实验室使用这些细胞对测试化合物进行高通量筛选。然而，目前的 hiPSC-CM 筛选往往针对表现出不成熟性和异质性的单个或少量细胞。此外， 心律失常事件仅限于早期后除极 (EAD) 或不规则搏动。 TdP 最终是 由折返或多灶性异位活动引起的多细胞组织现象，需要最少的 尺寸待揭晓。 该项目的目标是开发适合测试药理化合物和药物的体外模型 评估组织水平心律失常的风险。霍普金斯大学的三个研究实验室将贡献他们的专业知识 项目。用于支持 hiPSC-CM 的组织平台将是 Tung 开发的工程心脏切片 心脏电生理学实验室。生理功能改善的切片2.0版本即将推出 使用经过验证和推定的成熟兴奋剂，然后用于测试药物引起的折返或多灶性 心律失常。为此，将通过 Boheler 实验室的专业知识来追求细胞的代谢成熟， 该公司已将 CD36 确定为细胞表面标记，可区分代谢更成熟的细胞。一个 Kwon 心脏发育生物学实验室开发的生物信息学方法将用于遵循 心肌细胞的成熟轨迹。这些技术的融合将带来先进的 体外组织模型，将用于独立和机械地评估促心律失常作用 选择已被临床鉴定为低、中、高风险的化合物，并确定 心肌细胞的成熟状态是否影响其药物反应。这种方法代表了一种新的 心脏安全测试的范例——一种更接近预测 TdP 类心律失常事件的范例 心肌 – 并可能为 hiPSC-CM 的实用性、有效性和成熟依赖性建立新标准 作为评估心律失常风险的实验模型。