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（由申请人提供）：开发基于人类心肌细胞的预测心脏毒性的全光学测定法。心脏毒性是药物化合物所有阶段和限制的安全驱动戒断的主要原因。目前的心脏安全范例非临床指导 ICH S7B 侧重于体外 hERG 测定，事实证明，该测定在预测化合物对人体的心脏毒性方面效果不佳。在新药发现成本高、生产率低的背景下，更好的心脏安全性评估成为一个重要的公共卫生问题。 CSRC、HESI 和 FDA 正在开发一种新的心脏安全范例：综合体外致心律失常测定 (CiPA)，其中包括表征化合物在 iPSC 衍生的人心肌细胞中的电生理效应。 Q-State 的平台可以比目前的膜片钳检测更高的通量、更低的成本和更高的信息内容（多模态：电压、Ca2+、pH 和 ATP）来探测此类细胞的电生理学，并且比其他光学筛选工具具有更好的时间和空间分辨率以及更低的光毒性。在 68 万美元的 NINDS SBIR 第一阶段奖项和天使资金的支持下，Q-State Biosciences 正在开发一种专有的统包仪器和软件系统 (Optopatch)，用于同时对神经元膜电压进行光学扰动和光学测量。我们建议采用 Optopach 进行心肌细胞测量，并开发和验证 Optopatch 测定法，用于检测人 iPSC 衍生心肌细胞中药物引起的心脏毒性。目标 1. 分子工具。通过比较启动子、运输基序和基因传递方法，选择最有效的执行器（将蓝光转换为起搏的电刺激）和报告器（将 AP 波形转换为近红外荧光信号），并彻底表征分子工具的灵敏度、速度、光稳定性和可重复性。目标 2. 仪器仪表。采用 Optopatch 硬件和软件，同时对超过 1,000 个心肌细胞进行光学刺激和记录。开发流体和环境控制；修改软件以支持光学起搏、图像分割和心脏相关 AP 参数的计算。目标 3. 测定开发和测试。优化急性和慢性心脏毒性的测定。通过量化一组具有已知机制和心脏安全性的药物的 AP 变化来验证测定。我们将在先导化合物识别和优化阶段向药物发现公司提供非 GLP 心脏毒性筛选服务，补充并有可能最终取代 hERG 检测。我们还将向学术界提供 Optopatch-cardio 仪器和试剂；实现机理研究并拓宽电生理学平台的应用。