Develop predictive human cardiomyocyte-based all optical assay for cardiotoxicity

开发基于人类心肌细胞的心脏毒性预测全光学测定法

• 批准号: 8832817
• 负责人: Graham Thomas Dempsey
• 金额: $ 22.17万
• 依托单位: Q-STATE BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-11-15 至 2015-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8832817
• 关键词: Action Potentials; Acute; Address; Adoption; Adverse effects; Award; Biological Assay; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cells; Chronic; Communities; Computer software; Development; Drug Compounding; Drug Delivery Systems; Electrophysiology (science); Environmental Health; Evaluation; Fluorescence; Funding; Gene Delivery; Genes; Healthcare; Heart; Human; In Vitro; Institutes; Lead; Light; Manuals; Marketing; Measurement; Measures; Membrane; Methods; Modality; Molecular; National Institute of Neurological Disorders and Stroke; Neurons; Optics; Patients; Pharmaceutical Preparations; Phase; Phototoxicity; Productivity; Protocols documentation; Public Health; Reagent; Regimen; Reporter; Research; Resolution; Risk; Safety; Services; Signal Transduction; Small Business Innovation Research Grant; Solutions; Speed; Staging; Stimulus; System; Technology; Testing; Therapeutic; Time; Torsades de Pointes; United States Food and Drug Administration; Variant; Verapamil; Withdrawal; assay development; base; cellular imaging; cost; drug candidate; drug discovery; experience; imaging Segmentation; induced pluripotent stem cell; innovative technologies; instrument; instrumentation; millisecond; patch clamp; prevent; product development; promoter; public health relevance; screening; software systems; success; therapeutic development; tool; trafficking; voltage

 DESCRIPTION (provided by applicant): Develop predictive human cardiomyocyte-based all optical assay for cardiotoxicity. Cardiotoxicity is the leading cause of safety-driven withdrawal a all stages and limitation of drug compounds. The current cardiac safety paradigm nonclinical guidance ICH S7B focuses on in vitro hERG assays, which prove to be suboptimal in predicting a compound's cardiotoxicity in human. In the context of high cost and low productivity for new drug discovery, better cardiac safety evaluation becomes a significant public health issue. CSRC, HESI, and FDA are developing a new cardio safety paradigm: Comprehensive In vitro Proarrhythmia Assay (CiPA), which comprises characterizing electrophysiological effects of compounds in iPSC-derived human cardiomyocytes. Q-State's platform can probe electrophysiology of such cells with higher throughput, lower cost, and higher information content (multi-modalities: voltage, Ca2+, pH, and ATP) than current patch clamp assays, and with better temporal and spatial resolution and lower phototoxicity than other optical screening tools. Supported by a $680K NINDS SBIR Phase I award and angel funding, Q-State Biosciences is developing a proprietary turnkey instrument and software system (Optopatch) for simultaneous optical perturbation and optical measurement of membrane voltage in neurons. We propose to adapt Optopach for cardiomyocyte measurements and to develop and validate an Optopatch assay for drug-induced cardiotoxicity in human iPSC- derived cardiomyocytes. Aim 1. Molecular tools. Select the most effective actuators (converting blue light into electrical stimuli for pacing) and reporters (converting AP waveforms into near infrared fluorescence signals) by comparing promoters, trafficking motifs and gene delivery methods, and to thoroughly characterize the sensitivity, speed, photostability, and repeatability of the molecular tools. Aim 2. Instrumentation. Adapt Optopatch hardware and software to optically stimulate and record from > 1,000 cardiomyocytes simultaneously. Develop fluidics and environmental controls; modify software to support optical pacing, image segmentation, and calculation of cardiac-relevant AP parameters. Aim 3. Assay development and testing. Optimize assays for acute and chronic cardiotoxicity. Validate the assay by quantifying AP changes under a panel of drugs with known mechanisms and cardiac safety profiles. We will provide non-GLP cardiotoxicity screening services to drug discovery companies at the lead identification and optimization stages, supplementing, with the potential to eventually replace, hERG assays. We will also make Optopatch-cardio instrumentation and reagents available to the academic community; to enable mechanistic studies and broaden the applications of electrophysiology platform.

 描述（由申请人证明）：基于人类心肌细胞的所有光学关联新的药物发现，更好的心脏安全性成为CSRC，HESI和FDA的重大问题。 - 模式：电压，Ca2+和ATP比当前的贴片夹测定，并且具有Q-State Biosciences支持的其他光学筛选工具提议对心肌细胞测量，并在人IPSC偏离的心肌细胞中开发和验证TCH分析。比较启动子，流量基序和基因OD，并彻底表征分子工具的敏感性，速度和重复性。 ，并计算与心脏相关的AP参数。领先的识别和优化阶段，有可能取代事件，我们还将为学术界提供Optopatch-Cardio仪器和试剂；