Identification of Biomarkers of Cardiotoxicity using Metabolomics of Human Pluripotent Stem Cell-Derived Cardiomyocytes

利用人多能干细胞来源的心肌细胞的代谢组学鉴定心脏毒性的生物标志物

• 批准号: 9149275
• 负责人: ROBERT E BURRIER
• 金额: $ 51.15万
• 依托单位: STEMINA BIOMARKER DISCOVERY, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9149275
• 关键词: Accounting; Adverse drug effect; Adverse effects; Animal Model; Animals; Biological; Biological Assay; Biological Markers; Biotechnology; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cause of Death; Cell Line; Cells; Centers for Disease Control and Prevention (U.S.); Chemicals; Clinical; Data; Data Set; Development; Direct Lytic Factors; Drug Industry; Drug usage; FDA approved; Fibroblasts; Generations; Goals; Health; Human; In Vitro; Investigational Drugs; Marketing; Mass Spectrum Analysis; Measurement; Measures; Metabolic; Metabolism; Methods; Modeling; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Pluripotent Stem Cells; Preparation; Process; Public Health; Research; Safety; Sampling; Sensitivity and Specificity; Services; Small Business Innovation Research Grant; Staging; Statistical Data Interpretation; System; Technology; Testing; Time; Toxicology; Training; Undifferentiated; United States; Withdrawal; abstracting; assay development; base; biomarker discovery; biomarker identification; blind; candidate marker; cost; cost effective; cytotoxicity; data acquisition; developmental toxicity; drug candidate; drug development; drug discovery; drug market; high throughput screening; improved; in vitro Assay; in vitro Model; in vivo; induced pluripotent stem cell; metabolomics; novel strategies; novel therapeutics; pre-clinical; predictive modeling; response; safety testing; screening; small molecule; success; targeted biomarker; therapeutic development; tool

 DESCRIPTION (provided by applicant) Identification of Biomarkers of Cardiotoxicity using Metabolomics of Human Pluripotent Stem Cell- Derived Cardiomyocytes Project Summary/Abstract 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 improve public health, as well as alleviate the financial burden of compound attrition due to cardiotoxicity, through development of an in vitro assay to predict a compound's cardiotoxicity potential. To accomplish this, Stemina Biomarker Discovery ("Stemina") proposes to use metabolomics of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) exposed to known human cardiotoxic and non-cardiotoxic compounds. These technologies will be used to discover human, endogenous metabolite biomarkers which predict general cardiotoxicity as well as the specific type of cardiotoxicity (e.g., functional, structural). The use of metabolomics to measure small molecules secreted by hiPSC-CMs in response to compound exposure is a novel approach for evaluating cardiotoxicity and may pave the way for a new generation of more accurate, predictive toxicology screens using human cells. Stemina already used such a paradigm to complete the Phase I SBIR Application (1R43GM100640-01), as well as developed predictive methods to assess developmental toxicity potential in undifferentiated pluripotent stem cells (devTOX(tm)). Stemina's long-term goal is to develop a human cell- based, high-throughput cardiotoxicity screen as a valuable tool to pharmaceutical, biotech, and agrichemical companies during early development of therapeutics and chemicals. In order to achieve this goal, Stemina first proposes to develop an optimized and reproducible experimental platform to evaluate spent media collected from hiPSC-CMs (aim 1). We will evaluate various sample preparation methods, LC-MS columns and conditions, and perform robustness testing in order to establish the most reproducible measurement of the complete set of secreted metabolites, or secretome, in hiPSC-CMs using our system. In aim 2, we will use the above platform to evaluate spent media from hiPSC- CMs response to a training set of 60 compounds consisting of functional-, structural-, general-, and non- cardiotoxicants to establish a predictive metabolomic model. 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. The data acquired here will be used to establish a predictive metabolic signature indicative of general cardiotoxicity and specific type of cardiotoxicity (e.g., functional, structural). Stemina will then test the performace of the predictive model(s) of biomarker signature(s) on a test set of 20 compounds. In aim 3, Stemina will confirm the structural identity of the predictive metabolites and evaluate their biological significance as confirmed biomarkers. Lastly in aim 4, a targeted biomarker assay will be developed using targeted LC-MS methods that measure the confirmed biomarkers. Further, the ability of our biomarkers to adequately predict cardiotoxicity will be tested through the use o a blind study comprised of compounds acquired from partnering companies. Completion of these aims will enable the development of a commercial assay able to detect the validated biomarkers of cardiotoxicity, similar to an existing test Stemina currently markets for developmental toxicity (devTOX(tm) quickPredict). Stemina will subsequently utilize this assay to market a service capable of predicting whether a compound will induce cardiotoxicity and serve pharmaceutical companies in preclinical screening trials. Such a service provides the first human cell-based screening assay for cardiotoxicity founded on cardiomyocyte metabolism.

 描述（由申请人提供） 使用人多能干细胞来源的心肌细胞的代谢组学鉴定心脏毒性的生物标志物 项目摘要/摘要 心脏安全性是制药行业化合物消耗和 FDA 批准的药物退出市场的主要原因之一。该提案的目的是通过开发体外测定来预测化合物的心脏毒性潜力，从而改善公众健康，并减轻因心脏毒性而导致的化合物消耗的经济负担。为了实现这一目标，Stemina Biomarker Discovery（“Stemina”）建议使用暴露于已知人类心脏毒性和非心脏毒性化合物的人类诱导多能干细胞衍生心肌细胞（hiPSC-CM）的代谢组学。这些技术将用于发现人类内源性代谢物生物标志物，预测一般心脏毒性以及特定类型的心脏毒性（例如功能性、结构性）。使用代谢组学来测量 hiPSC-CM 响应化合物暴露而分泌的小分子是一种评估心脏毒性的新方法，并可能为使用人体细胞进行新一代更准确、预测性毒理学筛查铺平道路。 Stemina 已经使用这样的范例来完成 I 期 SBIR 申请 (1R43GM100640-01)，并开发了评估未分化多能干细胞 (devTOX(tm)) 发育毒性潜力的预测方法。斯泰米纳的 长期目标是开发基于人体细胞的高通量心脏毒性筛查，作为制药、生物技术和农化公司在治疗药物和化学品早期开发过程中的宝贵工具。为了实现这一目标，Stemina 首先建议开发一个优化且可重复的实验平台来评估从 hiPSC-CM 收集的废培养基（目标 1）。我们将评估各种样品制备方法、LC-MS 色谱柱和条件，并进行稳健性测试，以便使用我们的系统对 hiPSC-CM 中的全套分泌代谢物或分泌组进行最具重复性的测量。在目标 2 中，我们将使用上述平台来评估 hiPSC-CM 对由功能性、结构性、一般性和非心脏毒物组成的 60 种化合物训练集的反应，以建立预测代谢组学模型。其丰度取决于细胞是否用心脏毒性诱导剂或非诱导剂处理而变化的小分子将作为心脏毒性的候选生物标志物。这里获取的数据将是 用于建立指示一般心脏毒性和特定类型心脏毒性（例如功能性、结构性）的预测代谢特征。 Stemina 随后将在 20 种化合物的测试集上测试生物标志物特征的预测模型的性能。在目标 3 中，Stemina 将确认预测代谢物的结构特性，并评估其作为已确认生物标志物的生物学意义。最后，在目标 4 中，将使用有针对性的 LC-MS 方法开发有针对性的生物标志物测定，以测量已确认的生物标志物。此外，我们的生物标志物充分预测心脏毒性的能力将通过使用从合作公司获得的化合物组成的盲法研究进行测试。完成这些目标将能够开发出一种商业化验方法，能够检测经验证的心脏毒性生物标志物，类似于 Stemina 目前销售的发育毒性现有测试 (devTOX(tm) QuickPredict)。 Stemina 随后将利用这种检测方法来营销一项能够预测化合物是否会引起心脏毒性的服务，并为制药公司的临床前筛选试验提供服务。此类服务提供了第一个基于人体细胞的基于心肌细胞代谢的心脏毒性筛查测定。