Maturation of stem cell-derived cardiomyocytes using a simulated microgravity bioreactor and combinations of extracellular developmental cues

项目摘要 药物在开发过程中的失败是非常昂贵和危险的。事实上， 开发一种药物需要10年以上的时间和20 - 30亿美元，其中大部分费用来自于 失败的候选人。大多数失败发生在开发过程的最后，此时成本最高 以及药物接触到大多数患者的时间。失败的最常见原因之一是化合物的 导致患者心律失常的倾向。在一些罕见的情况下，这些心脏毒性作用甚至不 在临床试验中检测到，并且只有在药物暴露于大众时才被发现， 导致对患者的伤害和从市场上昂贵的退出。因此，FDA 要求所有新药都要进行心脏毒性测试，但他们和制药行业都意识到， 筛选工具不足。这导致了越来越多的筛选工具的市场，这些工具的预测性比 现有技术。人诱导多能干细胞衍生的心肌细胞（hPSC-CM）代表了一种新的心肌细胞。 为临床前药物筛选建立具有高度代表性的体外组织模型提供了一条有前途的途径。 然而，大多数hPSC-CM模型不发育成成熟的成体样组织，因此不能重现一些细胞因子。 体内药物反应。NanoSurface Biomedical正在申请第1阶段SBIR资金，以开发组合 一种提高心脏干细胞成熟以改善药物诱导的心脏毒性筛选的方法 基于模拟微重力生物反应器的干细胞成熟。这种方法会产生组织 其在功能上更成熟，并且可以被进料到各种下游测定中。我们假设 模拟微重力、细胞图案化、电刺激和代谢底物的组合将 改善心脏结构和功能发育，以收集更多的心脏毒性数据 预测能力。为了验证这些假设，本项目将集中在协议和线索的优化， 本发明还提供了可以可再现地且可靠地使hPSC-CM成熟的组合。该公司将制定协议， 验证了这些组合线索可以增强体外hPSC-CM成熟的假设。成熟意志 通过一套结构，电生理和功能指标结合统计学进行评估 分析（目标1）。最有效的成熟方案将用于生成hPSC-CM，用于使用 下游药物毒性测定（目的2）。这些数据将用于评估该方法的有效性， 最终在第2阶段扩大规模，用于开发高度预测性的体外心脏试验， 第三阶段的商业发布和市场交付。成功验证公司的组合 该方法将产生一种创新的新产品，旨在缓解临床前毒性的关键缺陷 模型，并减少药物开发过程中的成本和时间。

PROJECT SUMMARY The failure of a drug during the development process is extraordinarily costly and dangerous. Indeed, it can take upwards of 10 years and $2-3 billion to develop just one drug, and much of that cost is incurred from the candidates that fail. Most failures occur towards the end of the development process when the costs are highest and when the drug is exposed to the most patients. One of the most common reasons for failure is a compound’s propensity for causing cardiac arrhythmias in patients. In some rare cases, these cardiotoxic effects aren’t even detected during clinical trials and are only discovered once the drug is exposed to the population at large, resulting in harm to patients and a costly withdrawal from the market place. Consequently, the FDA has mandated that all new drugs be tested for cardiotoxic effects, but they and the drug industry realize that current screening tools fall short. This has led to a growing market for screening tools that are more predictive than existing technologies. Human induced pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) represent a promising avenue towards building high-representative in vitro tissue models for preclinical drug screening. However, most hPSC-CM models do not develop into mature, adult-like tissue and thus fail to recapitulate some in vivo drug responses. NanoSurface Biomedical is applying for Phase 1 SBIR funding to develop a combinatorial approach to enhance the maturation of cardiac stem cells for improved drug-induced cardiotoxicity screening based on a simulated microgravity bioreactor-based maturation of stem cells. The approach will generate tissues that are more functionally mature and can be fed into various down-stream assays. We hypothesize that the combination of simulated microgravity, cell patterning, electrical stimulation and metabolic substrates will improve cardiac structural and functional development to enable the collection of cardiotoxicity data with more predictive capacity. To test these hypotheses, this project will focus on the optimization of protocols and cues in combinations that can reproducibly and reliably mature hPSC-CMs. The company will develop protocols and validate the hypothesis that these combinatorial cues can enhance hPSC-CM maturation in vitro. Maturation will be assessed via a suite of structural, electrophysiological, and functional metrics combined with statistical analysis (Aim 1). The most effective maturation protocol will be used to generate hPSC-CM for validation using downstream drug toxicity assays (Aim 2). These data will be used to assess the validity of the approach for eventual scale up during Phase 2 for the development of highly predictive in vitro cardiac assays, and for commercial release and market delivery in Phase 3. Successful validation of the company’s combinatorial approach will produce an innovative new product aimed at relieving critical deficiencies in preclinical toxicity models and reduce cost and time in the drug development process.