High-throughput nanoMEA-based Proarrhythmia Assay

 描述（由申请人提供）：在临床前和临床开发后期阶段对新化学实体进行鉴定是一项极其昂贵的事件，最常见的是与检测到新药中的非预期致突变特性相关。未被发现的疟疾诱导作用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进行潜在的致瘤性测试，这导致了准确且具有成本效益的临床前筛选工具的市场不断增长。人多能干细胞衍生的心肌细胞（hPSC-CM）代表了产生用于此类应用的上级体外心脏组织的手段。然而，hPSC-CM不能发育成成人心肌组织的适当代表是在有效的临床前筛选方案中使用这些细胞构建体的主要障碍。准确再现成人心肌的形式和功能的成熟心脏组织的产生对于提供能够可靠地预测化合物在转移到临床环境时的功效和/或毒性的临床前数据是必要的。该提案的重点是开发纳米图案化微电极阵列（MEA）平台，以高通量和预测方式评估新型化合物的促炎潜力。已知纳米形貌表面促进培养的hPSC-CM向代表成人心肌的表型成熟。我们假设，这种地形信号线索与MEA的整合将使这些成熟的人心肌细胞在体外的电生理性能的分析。此外，我们认为，地形介导的成熟的hPSC-CM将导致产生的心脏单层的电生理特性更密切地代表成人心脏组织的传导速度，各向异性传导模式，和场电位持续时间。最后，我们认为，建立这种成熟的功能测定将使收集的化合物促性腺激素数据具有更大的预测能力，在体外重建体内药物作用。为了验证这些假设，本基金将集中于地形图案化多孔膜电极的设计和生产（任务1）。然后将研究在该平台内培养的hPSC-CM产生已知致心律失常化合物的药物诱导的心律失常数据的能力，所述致心律失常化合物代表这些药物的体内活性（任务2）。将化合物对纳米图案化和平坦心肌细胞单层的作用与体内药物活性进行比较，以证明我们的平台在预测心肌对药物治疗的反应方面所提供的改进。这种纳米图案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有效性，并有可能简化当前的药物开发。该系统，我们拥有的知识产权，将是相当大的兴趣，学术和药物筛选实验室，我们将寻求商业化我们的原型成功完成后，这笔赠款。

 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.