A phenotypic screen for osteoarthritic pain therapeutics using all-optical electrophysiology.

项目概述：骨性关节炎疼痛疗法的表型筛查使用ALL- 光电生理学 慢性疼痛影响着美国超过1亿成年人，其中许多人找不到 用目前的药物缓解症状。在这些患者中，近三分之一患有骨性关节炎。 目前治疗慢性疼痛的方法包括阿片类药物和非类固醇炎药。 然而，这些药物在慢性治疗中的疗效受到药物发展的限制。 耐受性和剂量限制毒性。尤其是阿片类药物具有高度的成瘾性和依赖性 已经造成了巨大的社会负担。尽管明确的、未得到满足的医疗需求和重要的 在研究活动中，几乎没有针对慢性疼痛的新的非阿片类药物出现 过去十年。 Quell Treateutics使用Optopatch平台进行全光电生理 以足以进行表型筛选的吞吐量对神经元进行测量。使用工程设计 光遗传蛋白、蓝光和红光可以用来刺激和记录神经元的活动， 分别进行了分析。定制显微镜可以记录100英尺高的S的电生理 与高灵敏度和时间分辨率并行的神经元，这一能力目前还不能 可与任何其他平台筛选技术一起使用。 在这里，我们将Optopatch平台与慢性疼痛的体外模型相结合，其中背部 根神经节(DRG)感觉神经元沐浴在一种发现的炎症介质的混合物中 在骨性关节炎患者的关节中。炎性合剂对神经元的作用 变得极度兴奋，模仿预期的细胞疼痛反应。我们计算出 关节炎疼痛的功能表型，反映了动作电位形状的差异 以及对不同刺激做出反应的放电频率。我们将筛选出小分子化合物 逆转疼痛表型，同时最大限度地减少与疼痛表型垂直的神经元行为的扰动 疼痛表型，体外“副作用”。我们将反筛选来自 来自心脏的大脑皮层和心肌细胞优先考虑选择性作用于 发炎的感觉神经元。排名最高的化合物将经过化学优化和 它们的药代动力学、药物代谢和体内疗效将被表征。我们的目标是 先进的疼痛治疗发现，这最终可能有助于缓解美国的阿片类药物危机。

Project Summary: A phenotypic screen for osteoarthritic pain therapeutics using all- optical electrophysiology Chronic pain affects over 100 million adults in the United States, many of whom cannot find relief with current medications. Of these patients, nearly 1/3 of them suffer from osteoarthritis. Current treatments for chronic pain include opioids and non-steroidal inflammatory agents. However, efficacy of these drugs in chronic treatment is restricted by the development of tolerance and dose-limiting toxicities. Opioids in particular are highly addictive, and dependency has caused a large societal burden. Despite the clear, unmet medical need and significant research activity, few new classes of non-opioid drugs targeting chronic pain have appeared in the past decade. Quell Therapeutics uses the Optopatch platform for making all-optical electrophysiology measurements in neurons at a throughput sufficient for phenotypic screening. Using engineered optogenetic proteins, blue and red light can be used to stimulate and record neuronal activity, respectively. Custom microscopes enable electrophysiology recordings from 100’s of individual neurons in parallel with high sensitivity and temporal resolution, a capability currently not available with any other platform screening technology. Here, we combine the Optopatch platform with an in vitro model of chronic pain, where dorsal root ganglion (DRG) sensory neurons are bathed in a mixture of inflammatory mediators found in the joints of osteoarthritis patients. The neurons treated with the inflammatory mixture become hyperexcitable, mimicking the anticipated cellular pain response. We calculate the functional phenotype of arthritis pain, which captures the difference in action potential shape and firing rate in response to diverse stimuli. We will screen for small molecule compounds that reverse the pain phenotype while minimizing perturbation of neuronal behavior orthogonal to the pain phenotype, the in vitro “side effects.” We will counter-screen against neurons from the cerebral cortex and cardiomyocytes from the heart to prioritize compounds that act selectively in inflamed sensory neurons. The highest ranking compounds will be chemically optimized and their pharmacokinetic, drug metabolism, and in vivo efficacy will be characterized. Our goal is to advance therapeutic discovery for pain, which may ultimately help relieve the US opioid crisis.