High-throughput nanoMEA-based Proarrhythmia Assay

项目总结 新药在开发后期失败是一件代价极其高昂的事件，通常与 检测新药中意想不到的致心律失常特性。未被检测到的心律失常诱发效应有 这也是药物退出市场的一个常见原因。因此，FDA现在要求所有新的 对药物进行潜在的致心律失常特性测试，这导致了对精确度和成本的日益增长的市场 有效的临床前筛查工具。人多能干细胞来源的心肌细胞(hPSC-CMS)为 为这种应用产生优良的体外心脏组织的手段。然而，hPSC-CMS无法 在标准的培养条件下发展成足够的成人心肌组织是一个主要的 阻碍这种细胞结构在有效的临床前筛查方案中的使用。成熟的一代 准确概括成人心脏的形式和功能的心脏组织是必要的，以提供 能够可靠地预测化合物在转移到临床时的有效性和/或毒性的临床前数据 布景。我们的第一阶段SBIR项目证明了纳米微电极阵列(纳米MEA)可以 用于促进心肌细胞成熟，达到更具代表性的药物反应的程度。 基于第一阶段取得的成果，NanoSurface Biomedical正在申请第二阶段SBIR资金，以 进一步开发和优化集成原型纳米MEA系统，以促进心脏成熟度和高 改进药物心脏毒性筛查的吞吐功能分析。我们假设 384孔纳米MEA平板的建立将促进心脏结构和功能的发展，以使 高通量化合物毒性数据的收集，具有更强的预测能力。为了测试这些 假设，此次拨款续期将集中在我们优化的高通量纳米MEA板的验证上 设计，包括建立定制的硬件和软件，以促进快速数据分析，如 以及制定用于设备验证的关键生物指标(目标1)。然后我们将利用这个平台来 研究hPSC-CMS对已知的致心律失常化合物产生功能反应的能力 代表了这些药物在体内的活性(目标2)。细胞系的可变性也将被调查以 了解基因类型差异是如何在体外转化为功能差异的。最后一点是， 纳米MEAs促进hPSC-CMS疾病表型从结构上的发展 将对心肌病患者进行调查，作为扩大我们最终产品用途的一种手段。这个 纳米粒子对心肌细胞的结构影响为这些地形图的能力提供了强有力的理论基础 帮助区分疾病表型与野生型对照的底物，并代表了第二个实质性的 这项技术的市场。我们的高通量纳米MEA系统的成功验证将产生 一种创新的新产品，专为缓解当前的严重缺陷和降低成本而设计 临床前药物开发流程。

PROJECT SUMMARY Failure of new drugs at late stages of development is an extremely costly event, commonly associated with the detection of unexpected arrhythmogenic properties in novel drugs. Undetected arrhythmia-inducing effects are also a common reason for drug withdrawal from the market. As a result, 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) offer 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 under standard culture conditions is a major impediment to the use of such cell-constructs in effective preclinical screening protocols. Generation of mature cardiac tissues that accurately recapitulate the form and function of the adult human heart is necessary to provide preclinical data capable of reliably predicting a compound’s efficacy and/or toxicity when transferred to a clinical setting. Our Phase 1 SBIR project demonstrated that nanopatterned microelectrode arrays (nanoMEAs) can be used to promote cardiomyocyte maturation to the point where more representative drug responses are achieved. Based on results achieved during Phase 1, NanoSurface Biomedical is applying for Phase 2 SBIR funding to further develop and optimize an integrated prototype nanoMEA system to enhance cardiac maturation and high throughput functional analysis for improved drug-induced cardiotoxicity screening. We hypothesize that the establishment of a 384-well nanoMEA plate will improve cardiac structural and functional development to enable the collection of high throughput compound toxicity data with greater predictive capacity. To test these hypotheses, this grant renewal will focus on the validation of our optimized high throughput nanoMEA plate design, including the establishment of custom-built hardware and software to facilitate rapid data analysis as well as development of key biological metrics for device validation (Aim 1). We will then use this platform to investigate the ability for hPSC-CMs to generate functional responses to known arrhythmogenic compounds that are representative of these drugs’ activity in vivo (Aim 2). Cell line variability will also be investigated to understand how genotypic differences translate into functional variance in vitro. Lastly, the capacity for nanopatterned MEAs to promote the development of disease phenotypes in hPSC-CMs from structural cardiomyopathy patients will be investigated as a means to broaden the utility of our eventual product. The structural impact of nanopatterns on cardiomyocytes provides strong rationale for the ability for these topographic substrates to help stratify disease phenotypes from wild type controls, and represents a second substantial market for this technology. Successful validation of our high throughput nanopatterned MEA system will produce an innovative new product designed specifically to relieve critical deficiencies and reduce cost in the current preclinical drug development process.