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

项目摘要 心脏病仍然是世界上主要的死亡原因，药物引起的心脏毒性是主要的 药物失效和退出临床或市场的原因，导致总体成功率较低 药物开发项目。体外心肌收缩能力模型有可能产生可预测的 早期数据，以减少因心脏毒性而导致的后期故障，并使发现和 开发有效的治疗方法更有可能在临床上成功转化。这样的心脏收缩能力 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 靶标不可知的收缩调节剂(如小分子、生物制品、RNAi)的发现 或者拯救健康的表型。同样，可以使用高通量的心肌收缩能力测试系统。 测试患者特异性诱导多能干细胞来源的心肌细胞(IPSC-CM)的可能治疗方法 以确定患者特定的“精准药物”治疗方法。不幸的是，没有一个单一的解决方案可以解决所有 终端用户的关键需求以及在现有技术中报告的机械终端、电子起搏 能力是以吞吐量为代价的。此STTR提案将提供一个集成的产品 通过将Forcell的FLECS收缩试验的优化版本与 建议的新型“Pace-Cap”电起搏系统设计用于标准井板格式，将 直接嵌入在微孔板盖上。在AIM中，一个24孔的盖内电子起搏机制的原型将 使用钙通量作为初步读数进行开发和验证。在Aim 2中，Forcell的收缩平台 将被开发成心脏微组织评估平台，由此产生的检测将被用于机械- 从功能上验证PACE-Cap。如果成功，这将代表着第一个一体化的心脏收缩能力 分析试剂盒既是高通量的，又具有内部起搏功能。第二阶段的工作将集中在扩展 将原型产能扩大到96口井，并制定快速生产战略。

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.