Engineered tissue-based, high-throughput compound profiling

描述（由申请人提供）：目前的NIGMS-SBIR资金支持InvivoSciences LLC（IVS）在2010年推出几条产品线。IVS通过销售三维（3D）细胞培养工具（MC-8 TM和IVS InsertsTM）获得收入，这些工具可以在没有任何支撑层的情况下生长各种水凝胶组织。培养工具使Palpator TM和Tissue Stretcher TM能够拉伸水凝胶组织进行生物力学测量和机械调节（例如，循环应力应用）。IVS还使用我们的工具和设备为工业和学术实验室提供合同研究服务，以分析化合物对细胞和组织生理学的影响。为了进一步证明我们筛选候选药物的能力，特别是对药物开发人员来说，市场需要针对其药理功能（包括毒性信息）已得到充分表征的化合物和药物进行基准研究。为了将我们目前的启动活动完全商业化，艾维斯将改进其快速药物筛选系统，该系统使用工程心脏组织（EHT）来监测测试化合物对心肌收缩力和相关调控分子的影响。在目标1中，将使用源自人诱导多能干细胞（iPS）的心肌细胞开发EHT，以使使用人样品的药物筛选系统商业化。使用这个系统，我们将确定一组药物的有益和毒性作用，基于药物诱导的EHT心脏功能的变化，以及作为其活动基础的信号转导途径。在目标2中，我们将制定ISO 13485：2003质量管理体系的具体要求，以便我们可以更有信心地为药物开发商提供合同研究服务。此外，使用16种已知的心脏有效和毒性药物/化合物的列表，我们将使用EHT测量药物诱导的心脏功能变化，以建立基准。在目标3中，我们将确定心脏毒性的机制，并证明EHT在体外预测心脏毒性的能力，而无需建立动物研究。我们的方法将推进药物靶点识别和优化以及生物标志物的发现，这对诊断心脏毒性至关重要。为了证明我们的方法能够阐明心脏毒性的机制，我们将使用shRNA和药物抑制mTOR（雷帕霉素的哺乳动物靶标）的基因敲除作为例子。我们的目标的成功完成将证明我们的体外系统预测药物诱导的人类心脏毒性的能力，显然有利于早期药物发现。

DESCRIPTION (provided by applicant): Current NIGMS-SBIR funding supported InvivoSciences LLC's (IVS) launch of several product lines in 2010. IVS generated revenues from the sales of three-dimensional (3D) cell culture tools (MC-8TM and IVS InsertsTM) that can grow various hydrogel tissues without any support layers. The culture tools enable Palpator TM and Tissue StretcherTM to stretch the hydrogel tissues for biomechanical measurements and mechanical conditioning (e.g., cyclic stress applications), respectively. IVS also performed contract research services, using our tools and devices, for industry and academic laboratories for profiling compound-induced effects on cell and tissue physiology. To further demonstrate our ability to screen drug candidates, especially for drug developers, the market demands benchmark studies against compounds and drugs whose pharmacological functions, including toxicity information, have been well-characterized. To fully commercialize our current start- up activities, IVS will improve its rapid drug screening system that uses engineered heart tissues (EHTs) to monitor the effects of test compounds on cardiac contractility and associated regulatory molecules. In Aim 1, EHTs will be developed using cardiomyocytes derived from human induced pluripotent stem (iPS) cells to commercialize a drug screening system using human samples. Using this system, we will determine the beneficial and toxic effects of a panel of drugs, based on the drug-induced changes in the cardiac functions of EHTs, as well as the signal transduction pathways that underlie their activities. In Aim 2, we will establish ISO 13485:2003-specified requirements for a quality management system so that we may more confidently provide contract research services for drug developers. In addition, using a list of 16 well-known cardio effective and toxic drugs/compounds, we will measure drug-induced cardiac function changes using EHTs to establish the benchmark. In Aim 3, we will identify mechanisms of cardiotoxicity, and will demonstrate the ability of the EHTs to predict cardiotoxicity in vitro, without the need for establishing animal studies. Our approach will advance drug target identification and optimization as well as biomarker discovery-critical for diagnosing cardiotoxicity. As a demonstration of the ability of our approach to elucidate a mechanism of cardiotoxicity, we will use, as an example, genetic knockdown with shRNA and drugs to inhibit mTOR (mammalian target of rapamycin). Successful completion of our aims will prove the ability of our in vitro system to predict drug-induced cardiotoxicity in humans, clearly benefiting early-stage drug discovery.