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.

项目概要 尽管许多神经系统疾病普遍存在且严重，但新疗法的开发 近 50 年来，各类药物一直低迷，这主要是由于缺乏良好的模型。 我们的目标是快速研究相关电和突触表型的系统和工具。 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病史相结合的第一个前景 使用快速体外技术可以研究这些细胞的复杂电生理行为。 使用 Q-State 最近开发的 Optopatch 平台和显微镜系统进行记录。 使用这些工具，可以同时刺激（蓝光）和记录（红光） 大约一百个神经元的电活动，时间分辨率为一毫秒， 该系统可用于单细胞空间分辨率和高信噪比测量。 单细胞兴奋性和放电模式或通过刺激子集来探测突触传递 展望未来，我们建议增加具有空间图案的蓝光的神经元。 在不牺牲功能的情况下提高显微镜吞吐量，并严格测试平台的 首先，显微镜将升级为先进的环境控制，96- 接下来是数据存储，以及孔板兼容性和用于化合物添加的流体处理机器人。 分析将安全地转移到云端，以处理 2.5 TB/天的数据速率。 显微镜功能齐全、灵敏度高、重现性好（孔与孔、板与板之间以及 最后，作为第一个应用，将使用对照化合物库进行测试。 我们将寻找一种稳健的、可筛选的 Dravet 综合征表型。 准备十名健康人和十名 Dravet 患者，寻找超越的放电差异 改善疾病表型的药物。 大多数细胞系是在临床上广泛有效的有希望的候选者。 经过验证的高通量电生理学平台，具有已确认的表型读数 人类 iPSC 疾病神经元有可能改变药物筛选格局 我们希望为这些可怕的神经疾病开辟一条新的治疗方法。 疾病。