Predictive assessment of acute and chronic cardiotoxicity using combinatorially matured hPSC-CMs

项目摘要 近90%的开发中药物未能进入市场。其中许多故障的发生是由于 心脏毒性在少数值得注意的案例中，一些药物通过了临床前筛选和临床试验，但却被取消了 一旦在大量患者群体中发现毒性作用，这些失败代表了 这是一个巨大的废物来源，并构成了将一种药物用于生产的约20亿美元成本的重要组成部分。 市场因此，FDA现在要求所有药物在上市前都要进行体外心脏毒性测试。 在人类身上测试。这导致了一个重要的和不断增长的市场，工具和技术，使更早 在接触患者之前检测毒性效应。然而，目前的筛查方法无法预测 药物在体内的作用方式;事实上，迫切需要更多的预测模型系统。此外，本发明还 大多数筛选侧重于急性毒性，而不测试长期结构毒性， 在病人长时间接触药物治疗后被发现。人诱导多能干细胞衍生 心肌细胞（hPSC-CM）是用于体外临床前毒性筛选的有吸引力的模型;它们相对 易于维护，来源于人体组织，并有可能减少对动物的需求。 实验然而，目前，基于hPSC-CM的测定法不能正确地复制细胞因子的功能。 人类的心脏这些细胞表现出与胎儿组织相似的表型，并且不像预期的那样对 已知作用的药物;在某些情况下，已知的不良作用药物不能在hPSC-CM中诱导毒性，而其他 仅在暴露于所述药物的超生理剂量时才显示出效果。药物发现 行业及其监管机构认识到hPSC-CM用于早期心脏毒性筛查的潜力，但也 了解到目前在药物开发过程中它们的使用有很大的局限性。因此，在本发明中， 很明显，准确再现体内药物反应的成熟心脏组织的产生 这是减少药物开发成本和浪费的重要机会。NanoSurface Biomedical， 股份有限公司、旨在应用生物工程方法提高hPSC-CM细胞的成熟度和预测能力 用于高预测性药物诱导的心脏毒性筛查。我们假设这些细胞会提供更多 急性和慢性毒性机制的体外心脏毒性检测的预测结果。我们将首先 重点是应用这些刺激并验证其预测毒性的能力（第1阶段）。经过验证后，我们 将描述这些细胞的表型，并将其用于各种旨在了解 在实验室中很难筛选的各种特定毒性机制（第2阶段）。我们将 使用这些数据来了解细胞成熟度在毒性检测中的作用，并为 全面的心脏毒性筛查框架。

PROJECT SUMMARY Nearly 90% of drugs under development fail to reach the market. Many of these failures occur due to cardiotoxicity. In a few notable cases, some drugs pass pre-clinical screens and clinical trials, only to be removed from the market once toxic effects are discovered in large patient populations. These failures represent a tremendous source of waste and constitute a significant part of the ~$2 billion cost of bringing a single drug to market. Consequently, the FDA now mandates that all drugs undergo in vitro cardiotoxicity testing before being tested in humans. This has led to a significant and growing market for tools and technologies that enable earlier detection of toxic effects before exposure to patients. However, current screening methods fall short of predicting how a drug will behave in the body; indeed there is a pressing need for more predictive model systems. Further, most screens focus on acute toxicity and do not test for longer-term structural toxicity which is typically only caught after a patient is exposed to the drug over long treatments. Human induced pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are an attractive model for in vitro preclinical toxicity screening; they are relatively easy to maintain, are derived from human tissue, and have the potential to reduce the need for animal experimentation. However, at present, hPSC-CM based assays do not properly replicate the function of the human heart. These cells exhibit phenotypes similar to that of fetal tissue and do not respond as expected to drugs of known effect; in some cases, known bad-actor drugs fail to induce toxicity in hPSC-CMs, while others only show effects when exposed to supra-physiological doses of the drug in question. The drug discovery industry and its regulators realize the potential of hPSC-CMs for early cardiotoxicity screening, but also understand that—at present—there are significant limitations to their use in the drug development process. Thus, it is clear that the production of mature cardiac tissues that accurately recapitulate in vivo drug responses represents a significant opportunity for reducing cost and waste in drug development. NanoSurface Biomedical, Inc., aims to apply bioengineering approaches to enhance the maturity and predictive power of hPSC-CM cells for highly predictive drug-induced cardiotoxicity screening. We hypothesize that these cells will give more predictive results in in vitro cardiotoxicity detection for both acute and chronic toxicity mechanisms. We will first focus on applying these stimuli and validating their ability to predict toxicity (Phase 1). After this validation, we will characterize the phenotypes of these cells and use them in a variety of assays targeted toward understanding a wide variety of specific toxicity mechanisms that are very difficult to screen in the laboratory (Phase 2). We will use these data to understand the role that cell maturity plays in toxicity detection and create a roadmap for a comprehensive cardiotoxicity screening framework.