Optopatch: high-throughput all-optical electrophysiology

Optopatch：高通量全光学电生理学

• 批准号: 9341395
• 负责人: Christopher Werley
• 金额: $ 31.8万
• 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: 2014-04-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9341395
• 关键词: Action Potentials; Affect; Afferent Neurons; Anticonvulsants; Automation; Award; Behavior; Biological Assay; Biological Models; Biological Sciences; Cell Line; Cells; Clinic; Clinical; Cloud Computing; Complex; Computer software; Custom; Data; Data Analyses; Data Storage and Retrieval; Databases; Development; Disease; Disease model; Drug usage; Electrophysiology (science); Engineering; Epilepsy; Fire - disasters; Funding; Genetic; Hippocampus (Brain); Hour; Human; In Vitro; Incubators; Individual; Libraries; Light; Liquid substance; Manuals; Measurement; Measures; Microscope; Modeling; Motion; Motor Neurons; Mus; Neurologic; Neurons; Noise; Optics; Output; Patients; Pattern; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase; Phenotype; Phototoxicity; Physiology; Positioning Attribute; Potassium Channel; Preclinical Drug Evaluation; Prevalence; Production; Property; Protein Engineering; Rattus; Recording of previous events; Reporter; Reproducibility; Resolution; Robot; Sampling; Secure; Severities; Shapes; Signal Transduction; Spinal Ganglia; Stem cells; Stimulus; Synapses; Synaptic Transmission; Synaptic plasticity; Syndrome; System; Technology; Testing; Therapeutic; Transcend; Variant; base; cell behavior; cell type; cloud based; cost; disease phenotype; genetic manipulation; induced pluripotent stem cell; instrument; millisecond; multidisciplinary; nervous system disorder; novel; novel drug class; optogenetics; phase 1 study; response; stem cell biology; temporal measurement; terabyte; tool; voltage

Project Summary In spite of the prevalence and severity of many neurological disorders, the development of new classes of drugs has been sluggish for 50 years. This is due largely to the lack of good model systems and tools to rapidly study relevant electrical and synaptic phenotypes. We aim to overcome these challenges. Recent advances in induced pluripotent stem cell (iPSC) technology reveal the first prospects for studying human neurons paired with clinical histories using fast in vitro technologies. The complex electrophysiological behavior of these cell can be recorded with the Optopatch platform and microscope systems recently developed at Q-State. With these tools, it is possible to simultaneously stimulate (blue light) and record (red light) electrical activity from around a hundred neurons with one millisecond temporal resolution, single cell spatial resolution, and high signal to noise. This system can be used to measure single cell excitability and firing patters or to probe synaptic transmission by stimulating a subset of neurons with spatially patterned blue light. Moving forward, we propose to increase microscope throughput without sacrificing capabilities and rigorously test the platform’s performance. First, the microscope will be upgraded with advanced environmental controls, 96- well plate compatibility, and a fluid-handling robot for compound addition. Next, data storage and analysis will be securely moved to the cloud to handle 2.5 Terabyte/day data rates. Once the microscope is fully functional, sensitivity and reproducibility (well to well, plate to plate, and batch to batch) will be tested using a library of control compounds. Finally, as a first application, we will search for a robust, screenable phenotype for Dravet syndrome. Neurons will be prepared from ten healthy and ten Dravet patients to look for differences in firing that transcend variation in the genetic background. A drug that ameliorates the disease phenotype in the majority of cell lines is a promising candidate to be broadly effective in the clinic. A well validated, high-throughput electrophysiology platform with confirmed phenotypic readouts in human iPSC disease neurons has the potential to change the drug screening landscape for neurological disorders. We hope to open a new path to finding treatments for these horrible diseases.

项目摘要 尽管许多神经系统疾病的患病率和严重程度，但新的发展 50年来，药物的类别一直迟钝。这主要是由于缺乏良好的模型 快速研究相关电气和突触表型的系统和工具。我们的目标 克服这些挑战。诱导多能干细胞（IPSC）的最新进展 技术揭示了研究人类神经元与临床历史配对的第一个前景 使用快速的体外技术。这些细胞的复杂电生理行为可以是 用Q-State最近开发的OptoPatch平台和显微镜系统记录。 使用这些工具，可以简单地刺激（蓝光）并记录（红灯） 来自大约一百个神经元的电活动，具有一毫升临时分辨率， 单细胞空间分辨率，高信号到噪声。该系统可用于测量 单细胞兴奋性和发射模式或通过刺激子集来探测突触传播 具有空间图案的蓝光的神经元。向前迈进，我们建议增加 显微镜吞吐量而无需牺牲功能并严格测试平台的 表现。首先，显微镜将通过高级环境控制升级，96- 孔板兼容性和用于复合添加的流体处理机器人。接下来，数据存储 分析将被牢固地移至云中，以处理2.5 trabyte/Day数据速率。一次 显微镜具有完全功能性，灵敏度和可重复性（井到板到板到板，然后 批次到批量）将使用控制化合物库进行测试。最后，作为第一个应用程序， 我们将为Dravet综合征寻找可靠的，可筛选的表型。神经元会 由十名健康和十名DRAVET患者制备，以寻找超越的射击差异 遗传背景的变化。一种改善疾病表型的药物 大多数细胞系都是有前途的候选人在诊所中广泛有效的。井 经过验证的高通量电生理学平台，并具有确认的表型读数 人IPSC疾病神经元有可能改变药物筛查格局 神经系统疾病。我们希望为寻找这些可怕的治疗方法打开新的途径 疾病。