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率的主要原因，而药物引起的心脏毒性是主要的 毒品衰竭和从诊所或市场退出的原因，导致整体成功率差 药物开发计划。心脏收缩的体外模型有可能产生预测性 在管道中的早期数据，以减少由于心脏毒性引起的后期失败，并启用发现和 开发有效的疗法更有可能在诊所成功翻译。这种心脏收缩 模型可以用作目标引导（拮抗）或 合同调制剂（例如，小分子，生物制剂，RNAi）的靶标不到静态发现 或营救健康的表型。同样，可以使用高通量心脏收缩性评估系统 测试患者特异性诱导的多能干细胞衍生的心肌细胞（IPSC-CM）的可能治疗方法 确定患者特定的“精度医学”治疗方法。不幸的是，没有一个解决方案解决所有问题 最终用户和现有技术报告机械终点的关键需求，电气起搏 功能是以吞吐量为代价的。该STTR提案将提供集成的产品 通过将Forcyte Flecs的收缩性评估的优化版本与A与A结合在一起来满足这些需求 拟议的新颖的“速度帽”电气加速系统，专为标准固定板格式而设计 直接嵌入在微板盖上。在AIM上，盖上电气起搏机构的24孔原型将 使用钙通量作为初步读数进行开发和验证。在AIM 2中，Forcyte的收缩力平台 将发展为心脏微组织评估平台，由此产生的测定将用于机械 在功能上验证速度帽。如果成功，这将代表第一个多合一心脏收缩率 分析套件既是高通量，又具有内部起搏功能。第2阶段的工作将重点放在扩展 原型吞吐量为96孔，并制定快速生产策略。