Combined Cardiomyopathy, e.g., of Cancer Chemotherapeutics, and Proarrhythmia for Cardiotoxicity Clinical Trials-in-a-Dish (CTiD) with iPSC-Derived Cardiomyocytes

联合心肌病，例如癌症化疗药物和致心律失常，使用 iPSC 来源的心肌细胞进行心脏毒性临床试验 (CTiD)

• 批准号: 10268102
• 负责人: Jeffrey H. Price
• 金额: $ 29.99万
• 依托单位: VALA SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10268102
• 关键词: Action Potentials; Address; Aging; American; Animal Experimentation; Animal Model; Animal Testing; Animals; Antineoplastic Agents; Arrhythmia; Artificial Intelligence; Biological Assay; Calcium; Cancer Survivor; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiotoxicity; Cardiovascular system; Cell Membrane Permeability; Cell membrane; Cells; Cellular Structures; Cessation of life; Clinic; Clinical; Clinical Trials; Complex; Contract Services; Contractile Proteins; Dangerousness; Development; Dose; Drug Industry; Female; Fluorescence; Goals; Gold; Growth; Guidelines; Heart; Heart Diseases; Hour; Human; Image; Image Cytometry; Impairment; In Vitro; Kinetics; Label; Lead; Malignant Neoplasms; Measures; Methods; Microscopy; Mitochondria; Motion; Myocardium; Myopathy; Outcome; Pathologic; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Population; Pre-Clinical Model; Production; Protocols documentation; Publications; Publishing; Quality Control; Recording of previous events; Recurrence; Reporting; Research Contracts; Risk Assessment; Robotics; Safety; Sales; Sarcoplasmic Reticulum; Science; Sensitivity and Specificity; Small Business Technology Transfer Research; Source; Specificity; Speed; Structure; System; Technology Transfer; Testing; Time; Tissue Donors; Toxic effect; Treatment Side Effects; anti-cancer therapeutic; base; calcium indicator; cancer recurrence; cancer survival; cancer therapy; cancer type; cardiovascular risk factor; cell injury; cellular imaging; chemotherapy; clinical predictors; compound 30; cost; cytotoxicity; dosage; drug candidate; drug development; drug discovery; drug testing; heart cell; high throughput screening; improved; induced pluripotent stem cell; insight; instrument; lead series; male; novel therapeutics; pre-clinical; pre-clinical research; response; scale up; screening; screening services; side effect; therapeutic candidate; tool; tumor; validation studies; voltage

Cardiotoxicity is a leading cause of drug discovery attrition across all of preclinical and clinical drug discovery. While the FDA and the Comprehensive in vitro Proarrhythmia Assay initiative (CiPA) are focused primarily on predicting proarrhythmic effects, drug attrition due to cardiomyopathy, or primary cardiac cytotoxicity, may be even more prevalent, is typically currently only carried out via animal studies, and limits dosage for many cancer chemotherapeutics. Due to improving cancer survival, it is increasing common for more cancer survivors of some cancer types to die of cardiac diseases due to cancer treatment side effects than cancer recurrence. Cardiac contractions are initiated by electrical depolarizations (action potentials, APs) that propagate through the heart and initiate calcium (Ca2+) transients that activate the contractile apparatus. Importantly, dysregulation of Ca2+ can trigger inappropriate early-after- and delayed-after- depolarizations (EADs and DADs) that initiate arrhythmias, inhibit mitochondrial function, and pathologically alter expression of contractile proteins. Chemotherapy and other drugs can also directly impair mitochondrial function, which is primarily thought to cause cytotoxicity, but can also cause arrhythmias. Cardiomyocytes are also heterogeneous in their voltage, calcium, and contractile functions, and in their responses to therapeutic candidates. Thus, it is highly desirable to simultaneously measure AP, Ca2+ and contractile function on a cell- by-cell basis, in human cardiomyocytes, but this is not possible with current test methods. To address this unmet need we propose to develop a high throughput (robotic) Kinetic Image Cytometry that simultaneously quantifes voltage, calcium, and contractile motion in cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs). The hiPSC-CMs will be labeled with fluorescent indicators of calcium and voltage, and the cells imaged via high-speed automated microscopy during contractile activity. The use of hiPSC-CMs will enable “clinical trials” in a dish, in which test compounds are tested across cells representing several donors. Phase I of this Fast-Track STTR project will develop the basic protocol and perform a proof-of-concept screen of 30 test compounds on hiPSC-CMs representing 5 donors. In Phase II, a large validation study (~350 compounds, 7-concentration dose-response, 30 min and 72 hr exposures) will be performed. Artificial intelligence will be utilized to optimize the sensitivity and specificity of the assay by detecting complex arrhythmia waveforms. This assay represents a human-based preclinical model that will be less expensive and more predictive for cardiotoxicity testing than animal models and will be marketed to the pharmaceutical industry for contract research.

药物毒性是所有临床前和临床药物发现中药物发现损耗的主要原因。 虽然FDA和综合体外致心律失常试验倡议（CiPA）主要关注 预测预防作用、心肌病引起的药物消耗或原发性心脏细胞毒性， 甚至更普遍的是，目前通常仅通过动物研究进行，并且限制了许多人的剂量。 癌症化疗药物。由于提高癌症生存率， 一些癌症类型的幸存者死于心脏病，由于癌症治疗的副作用比癌症 复发心脏收缩由电去极化（动作电位，AP）启动， 通过心脏传播并引发激活收缩装置的钙（Ca 2+）瞬变。 重要的是，Ca 2+的失调可以触发不适当的早期后去极化和延迟后去极化 (EADs和DAD），其引发心律失常，抑制线粒体功能，并病理性地改变 收缩蛋白化疗和其他药物也会直接损害线粒体功能， 主要被认为会导致细胞毒性，但也会导致心律失常。心肌细胞也是 在它们的电压、钙和收缩功能以及它们对治疗性药物的反应方面， 候选人因此，非常需要同时测量细胞上的AP、Ca 2+和收缩功能。 在人心肌细胞中，以细胞为基础，但这在目前的测试方法中是不可能的。为了解决这个 未满足的需求，我们建议开发一种高通量（机器人）动力学图像细胞仪，同时 定量电压，钙，和收缩运动的心肌细胞来源于人类诱导的多能 干细胞（hiPSC-CM）。hiPSC-CM将用钙和电压的荧光指示剂标记， 并在收缩活动期间通过高速自动显微镜对细胞成像。hiPSC-CM的使用 将在培养皿中进行“临床试验”，其中测试化合物在代表几种细胞的细胞中进行测试。 捐助者。这个快速通道STTR项目的第一阶段将制定基本协议并进行概念验证 在代表5个供体的hiPSC-CM上筛选30种测试化合物。在第二阶段，一项大型验证研究（约350 化合物，7浓度剂量-反应，30分钟和72小时暴露）。人工 智能将用于通过检测复杂的 心律失常波形这种检测方法代表了一种基于人类的临床前模型， 并且比动物模型更能预测心脏毒性测试，并将销售给制药公司。 工业合同研究。