Smart, integrated well-plates for ultra-high-throughput screening of excitation-contraction coupling in tissues

智能集成孔板，用于组织中兴奋-收缩耦合的超高通量筛选

• 批准号: 10267789
• 负责人: Ivan Pushkarsky
• 金额: $ 35万
• 依托单位: FORCYTE BIOTECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-14 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10267789
• 关键词: Address; Adhesions; Behavior; Biological; Biological Assay; Blood; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cells; Clinic; Complex; Coupling; Data; Development; Drug Side Effects; Electronics; Electrophysiology (science); Evaluation; Failure; Foundations; Geometry; Heart; Heart Diseases; Human body; In Vitro; Individual; Measurement; Measures; Mechanics; Microfabrication; Modality; Modeling; Output; Pathway interactions; Patients; Performance; Pharmaceutical Preparations; Phase; Phenotype; Production; Program Development; Pump; RNA Interference; Reporting; Research Personnel; Screening procedure; Signal Transduction; Small Business Technology Transfer Research; System; Techniques; Technology; Testing; Tissues; Translating; Validation; Withdrawal; Work; base; design; drug development; heart cell; heart function; high throughput screening; in vitro Model; induced pluripotent stem cell; microsystems; miniaturize; mortality; novel; precision medicine; printed circuit board; prototype; release of sequestered calcium ion into cytoplasm; response; scale up; screening; small molecule; success; technology development; therapeutically effective; voltage

Project Summary Heart disease remains the leading cause of mortality in the world and, drug-induced cardiotoxicity is a major cause of drug failure and withdrawal from the clinic or the market, contributing to the poor overall success rate of drug development programs. In vitro models of cardiac contractility have the potential to generate predictive data earlier in the pipeline to reduce later-stage failure due to cardiotoxicity, and enable discovery and development of effective therapeutics more likely to successfully translate in the clinic. Such cardiac contractility models could serve as valuable high-throughput phenotypic screening tools for target-guided (antagonistic) or target-agnostic discovery of contraction-modulating agents (e.g. small molecules, biologics, RNAi) that enhance or rescue the healthy phenotype. Similarly, a high-throughput cardiac contractility assay system could be used to test possible treatments in patient-specific induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) to identify patient specific “precision medicine” treatments. Unfortunately, no single solution address all of the key needs of the end-user and in existing technologies reporting mechanical endpoints, electrical pacing capabilities have come at the expense of throughput. This STTR proposal will deliver an integrated product addressing these needs by combining an optimized version of Forcyte’s FLECS contractility assay with a proposed novel “Pace-Cap” electrical pacing system designed for the standard well-plate format that will be embedded directly on the microplate lid. In aim, a 24-well prototype of the in-lid electrical pacing mechanism will be developed and validated using calcium flux as a preliminary readout. In aim 2, Forcyte’s contractility platform will be developed into a cardiac micro-tissue evaluation platform and the resulting assay will be used to mechano- functionally validate the Pace-Cap. If successful, this would represent the first all-in-one cardiac contractility assay kit that is both high-throughput and has internal pacing capabilities. Phase 2 work will focus on extending the prototype throughput to 96-wells and developing rapid production strategies.

项目摘要 心脏病仍然是世界上死亡的主要原因，药物引起的心脏毒性是主要的原因。 药物失败的原因和退出临床或市场，导致整体成功率低 药物开发项目。心脏收缩力的体外模型有可能产生预测 数据在管道中的早期，以减少后期失败，由于心脏毒性，并使发现和 开发更有可能在临床上成功转化的有效疗法。这种心脏收缩力 模型可以作为有价值的高通量表型筛选工具，用于靶向（拮抗）或 靶不可知的收缩调节剂（例如小分子、生物制剂、RNAi）的发现， 或拯救健康的表型。类似地，可以使用高通量心脏收缩性测定系统 测试患者特异性诱导多能干细胞衍生心肌细胞（iPSC-CM）的可能治疗方法 以确定患者特定的“精准医疗”治疗。不幸的是，没有一个单一的解决方案可以解决所有的问题。 最终用户的关键需求和现有技术中报告的机械终点、电起搏 能力是以吞吐量为代价的。该STTR提案将提供一个集成的产品 通过将Forcyte的FLECS收缩性测定的优化版本与 提出了一种新的“起搏帽”电起搏系统，设计用于标准孔板格式， 直接嵌入在微孔板盖上。在aim中，一个24孔的盖内电起搏机制原型将 使用钙通量作为初步读数进行开发和验证。在目标2中，Forcyte的收缩平台 将被开发成心脏微组织评估平台，并将使用所产生的测定来机械地 在功能上确认起搏帽。如果成功，这将是第一个多合一的心脏收缩力 这是一个高通量和具有内部起搏能力的检测试剂盒。第二阶段的工作将集中在 将原型产量提高到96口威尔斯井，并制定快速生产策略。