An all-optical electrophysiology platform for the discovery of pain therapeutics

项目摘要 在美国，慢性疼痛影响着超过1亿的成年人，治疗起来很有挑战性。 目前的治疗方法包括阿片类药物和非甾体类炎症剂。然而， 这些药物在长期治疗中受到剂量限制性毒性和长期阿片类药物的限制。 使用会导致依赖。尽管有明确的，未满足的医疗需求和重要的研究 活性，很少有药物靶向疼痛的基础上新的，非阿片类药物的机制已经出现在 过去十年Q-state已经创建了一个新的全光学平台（Optopatch）， 光遗传蛋白和定制显微镜，同时刺激和记录电 具有高灵敏度和时间分辨率的各种细胞类型的活性。我们集中我们 在基因验证的疼痛靶点SCN 9A（Nav1.7）上的努力，电压门控钠通道 这是感觉神经元传递疼痛信号所必需的。我们将应用Optopatch 技术有两种形式。1)使用Q-State化学文库的HTS筛选， 异源表达的Nav1.7通道测定，其复制生理加标活性。 针对其他Nav1.x通道的反筛选将以相同的检测格式进行， 确定化合物选择性。2)将以中等通量评价已鉴别的抑制剂 这些屏幕测量啮齿动物感觉神经元和人类iPS感觉神经元的兴奋性， 被炎症介质致敏。这套集成的检测试剂盒旨在 鉴定通过不同工作机制起作用的Nav1.7抑制剂，并优先考虑用于 使用感觉神经元功能的可缩放的体外模型进一步优化。这个平台将 作为一种有效的手段来选择化合物优化使用药物 化学和药代动力学、药物代谢和体内功效数据。

Project Summary Chronic pain affects over 100 million adults in the United States and is challenging to treat. Current treatments include opioids and non-steroidal inflammatory agents. However, efficacy of these drugs in chronic treatment is restricted by dose limiting toxicities, and prolonged opioid use can lead to dependency. Despite the clear, unmet medical need and significant research activity, few drugs targeting pain based on novel, non-opioid mechanisms have appeared in the past decade. Q-state has created a novel all-optical platform (Optopatch) using engineered optogenetic proteins and custom microscopes to simultaneously stimulate and record electrical activity from a variety of cell types with high sensitivity and temporal resolution. We focus our efforts on a genetically validated pain target, SCN9A (Nav1.7), a voltage-gated sodium channel that is required for pain signal transmission in sensory neurons. We will apply Optopatch technology in two formats. 1) An HTS screen of the Q-State chemical library using a heterologously expressed Nav1.7 channel assay that replicates physiological spiking activity. Counterscreens against other Nav1.x channels will be performed in the same assay format to determine compound selectivity. 2) Identified inhibitors will be evaluated in medium throughput screens that measure excitability in rodent sensory and human iPS sensory neurons that have been sensitized using inflammatory mediators. This integrated set of assays is designed to identify Nav1.7 inhibitors acting by diverse working mechanisms and prioritize compounds for further optimization using scalable in vitro models of sensory neuron function. This platform will be employed as an efficient means to select compounds for optimization using medicinal chemistry and pharmacokinetic, drug metabolism and in vivo efficacy data.