Identification of Biomarkers of Cardiotoxicity using Metabolomics of Human Plurip

描述（由申请人提供）：由于药物不良反应导致的心脏毒性是一个严重的问题，在患者接触之前尚未进行有效筛选。心脏安全性是制药行业化合物损耗的主要原因之一，也是fda批准的药物退出市场的主要原因之一。这项提议的目的是减轻由于心脏毒性引起的复合消耗的经济负担，但更重要的是，通过开发一种体外试验来预测药物诱导心脏毒性的能力，以改善公众健康。Stemina生物标志物发现（“Stemina”）建议通过代谢组学对来自人胚胎干细胞（hES）和人诱导多能干细胞（hiPS）细胞的处理心肌细胞进行研究。这些技术将用于发现人类内源性小分子生物标志物，预测心脏毒性，重点是心肌病。利用代谢组学来测量人类心肌细胞对药物反应时分泌的小分子是一种新颖的方法，可能为新一代更准确的预测毒理学筛选铺平道路。Stemina已经使用这种模式来开发预测方法来评估干细胞的发育毒理学。Stemina的长期目标是全面开发这种人性化、高通量的心脏毒性筛查，使其成为制药和生物技术公司在临床前开发治疗方法的宝贵工具。为了实现这一长期目标，Stemina首先提出为每个心肌细胞培养系统建立一个实验平台（Aim 1）。在Aim 2中，我们将使用这些系统为24种化合物（16种心脏毒性和10种非心脏毒性）中的每一种化合物开发剂量反应曲线，作为建立预测代谢组学模型的训练集。这些剂量反应曲线将用于确定Aim 3中使用的药物治疗浓度。然后，Stemina将建立药物诱导心脏毒性候选人类生物标志物的特定代谢特征（Aim 3）。为此，人类多能干细胞（hPS）衍生的心肌细胞将使用已知的心脏毒性诱导剂和非诱导剂进行治疗。用质谱法分析处理细胞的废培养基，以研究这些细胞的分泌代谢物或分泌组。小分子的丰度取决于细胞是否被心脏毒性诱导剂或非诱导剂处理，将作为心脏毒性的候选生物标志物。在未来的研究中，这些候选生物标志物将通过使用专门的质谱技术进行验证。最后，这些生物标志物充分预测心脏毒性的能力将通过使用盲法研究进行测试。这些目标的完成将鼓励与合作公司进一步讨论，以开发一种可以检测经过验证的心脏毒性生物标志物的试剂盒。然后，Stemina将利用该试剂盒推销一项服务，以预测一种化合物是否会诱发心脏毒性，这将为制药公司在临床前筛选试验中提供服务。这项服务将提供第一个基于心肌细胞代谢的人性化心脏毒性筛选试验，并可能改善公众健康。

DESCRIPTION (provided by applicant): Cardiotoxicity, as a result of adverse drug effects, is a serious problem that has yet to be effectively screened prior to patient exposure. Cardiac safety is one of the leading causes of compound attrition in the pharmaceutical industry and withdrawal of FDA-approved drugs from the market. The purpose of this proposal is to alleviate the financial burden of compound attrition due to cardiotoxicity, but more importantly, to improve public health through the development of an in vitro assay to predict a drug's ability to induce cardiotoxicity. Stemina Biomarker Discovery ("Stemina") proposes to do so through the use of metabolomics on treated cardiomyocytes derived from human embryonic stem (hES) and human induced pluripotent stem (hiPS) cells. These technologies will be used to discover human endogenous small molecule biomarkers which predict cardiotoxicity, with an emphasis on cardiomyopathy. The use of metabolomics to measure small molecules secreted by human cardiomyocytes in response to drugs is a novel approach and may pave the way for a new generation of more accurate predictive toxicology screens. Stemina has already used such a paradigm to develop predictive methods to assess development toxicology in stem cells. Stemina's long-term goal is to fully develop this humanized, high throughput cardiotoxicity screen so that it would be a valuable tool to pharmaceutical and biotech companies during preclinical development of therapeutics. In order to achieve this long term goal, Stemina is first proposing to establish an experimental platform for each cardiomyocyte culture system (Aim 1). In Aim 2, we will use these systems to develop a dose response curve for each of the 24 compounds (16 cardiotoxic and 10 non-cardiotoxic) as a training set to establish a predictive metabolomic model. These dose response curves will be used to determine 3 concentrations for drug treatment to be utilized in Aim 3. Stemina will then establish a specific metabolic signature of candidate human biomarkers of drug-induced cardiotoxicity (Aim 3). To do so, human pluripotent stem (hPS) cell-derived cardiomyocytes will be treated with well-established known inducers and non-inducers of cardiotoxiciy. The spent medium from treated cells will be analyzed with mass spectrometry in order to study the secreted metabolites, or secretome, of these cells. Small molecules whose abundances vary dependent upon whether cells were treated with an inducer or non-inducer of cardiotoxicity will serve as candidate biomarkers of cardiotoxicity. In future studies, these candidate biomarkers will be validated through the use of specialized mass spectrometry techniques. Lastly, the ability of these biomarkers to adequately predict cardiotoxicity will be tested through the use of a blind study. Completion of these aims will encourage further discussions with partnering companies in order to develop a kit that can detect the validated biomarkers of cardiotoxicity. Stemina will then utilize this kit to market a service to predict whether or not a compound will induce cardiotoxicity that will serve pharmaceutical companies in pre-clinical screening trials. Such a service will provide the first humanized screening assay for cardiotoxicity based on cardiomyocyte metabolism and will likely improve public health. PUBLIC HEALTH RELEVANCE: Adverse effects of drugs to patients are the fourth leading cause of death in the United States; one such an adverse effect is drug-induced cardiotoxicity, illustrating a need for a better predictive assay for cardiotoxicity. Stemina Biomarker Discovery proposes to develop biomarkers of cardiotoxicity that would drive a service model to predict the cardiotoxicity-potential of drug candidates in order to prevent cardiotoxicity in patients. Such a product will greatly improve public health.