High-throughput nanoMEA-based Proarrhythmia Assay

 描述(由申请人提供)：在临床前和临床开发的后期阶段，新化学物质的磨损是一项极其昂贵的事件，最常见的是在新药中检测到意外的致心律失常特性。未被发现的心律失常诱发效应也是药物退出市场的最常见原因。为此，FDA现在要求所有新药都要对潜在的致心律失常特性进行测试，这导致了对准确且具有成本效益的临床前筛查工具的市场不断增长。人多能干细胞来源的心肌细胞(hPSC-CMS)代表了为这类应用在体外产生优良心脏组织的手段。然而，hPSC-CMS不能发展成对成人心肌组织的充分表达是这些细胞构建在有效的临床前筛查方案中使用的主要障碍。为了提供临床前数据，能够可靠地预测化合物在转移到临床环境中的有效性和/或毒性，生成准确概括成人心肌形态和功能的成熟心脏组织是必要的。这项建议的重点是开发一个纳米修饰的微电极阵列(MEA)平台，利用该平台可以高通量和预测性地评估新化合物的致心律失常潜力。众所周知，纳米地形表面可以促进培养的hPSC-CMS向代表成人心肌的表型成熟。我们推测，这种地形图信号信号与MEAs的整合将能够在体外分析这些成熟的人心肌细胞的电生理性能。此外，我们假设hPSC-CMS的地形介导的成熟将导致心脏单层的产生，在传导速度、各向异性传导模式和场电位持续时间方面，具有更接近成人心脏组织的电生理特性。最后，我们认为，这一成熟的功能测定的建立将使收集的复合心律失常数据在体外重建体内药物效应方面具有更大的预测能力。为了验证这些假设，这笔赠款将重点放在设计和生产具有地形图案的多孔MEA(任务1)上。然后将研究在该平台内培养的hPSC-CMS为代表这些药物在体内的活性的已知致心律失常化合物产生药物引起的心律失常数据的能力(任务2)。纳米和扁平心肌细胞单层上的复合作用与体内药物活性的比较将被用来展示我们的平台在预测药物治疗的心肌反应方面所提供的改进。这种纳米MEA系统的成功验证将产生一种新产品，用于提高临床前药物筛选的效率，并有可能简化当前的药物开发。我们拥有这一系统的知识产权，它将引起学术和药物筛选实验室的极大兴趣，我们将在成功完成这笔拨款后寻求将我们的原型商业化。

 DESCRIPTION (provided by applicant): Attrition of new chemical entities at late preclinical and clinical stages of development is an extremely costly event, most commonly associated with the detection of unexpected arrhythmogenic properties in novel drugs. Undetected arrhythmia-inducing effects are also the most common reason for drug withdrawal from the market. To this end, the FDA now mandates that all new drugs be tested for potential arrhythmogenic properties, which has led to a growing market for accurate and cost effective preclinical screening tools. Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) represent the means to generate superior in vitro cardiac tissues for such applications. However, an inability for hPSC-CMs to develop into adequate representations of adult myocardial tissue is a major impediment to the use of these cell-constructs in effective preclinical screening protocols. Generation of mature cardiac tissues that accurately recapitulate the form and function of the adult human myocardium is necessary to provide preclinical data capable of reliably predicting a compound's efficacy and/or toxicity when transferred to a clinical setting. This proposal focuses on the development of a nanopatterned microelectrode array (MEA) platform with which to evaluate the arrhythmogenic potential of novel compounds in a high throughput and predictive manner. Nanotopographic surfaces are known to promote the maturation of cultured hPSC-CMs towards phenotypes representative of the adult human myocardium. We hypothesize that the integration of such topographic signaling cues with MEAs will enable the analysis of the electrophysiological performance of these matured human cardiomyocytes in vitro. Furthermore, we posit that the topography-mediated maturation of hPSC-CMs will lead to the generation of cardiac monolayers with electrophysiological properties more closely representative of adult cardiac tissue in terms of conduction velocity, anisotropic conduction patterns, and field potential durations. Finally, we suggest that the establishment of this mature functional assay will enable the collection of compound arrhythmogenesis data with greater predictive capacity in terms of recreating in vivo drug effects in vitro. To test these hypotheses, this grant will focus n the design and production (Task 1) of topographically patterned multiwell MEAs. The ability for hPSC-CMs cultured within this platform to generate drug-induced arrhythmia data for known arrhythmogenic compounds that are representative of these drugs' activity in vivo will then be investigated (Task 2). Comparison of compound action on nanopatterned and flat cardiomyocyte monolayers to drug activity in vivo will be used to demonstrate the improvement our platform offers in terms of predicting myocardial responses to drug treatment. Successful validation of this nanopatterned MEA system will produce a new product for advancing the efficacy of preclinical drug screening with the potential to streamline current pharmaceutical development. This system, for which we own the Intellectual Property, will be of considerable interest to both academic and pharmaceutical screening laboratories, and we will seek to commercialize our prototype upon successful completion of this grant.