PHARMACOLOGICAL MODULATION OF PIEZO1 CHANNELS

总结 机械敏感压电离子通道使真核细胞能够感知机械力。在 在脊椎动物中，两种压电同系物Piezo 1和Piezo 2在所有主要的生理功能中起着核心作用。 系统，并与许多疾病，包括高血压，干细胞症， 水肿、关节弯曲、炎症、疼痛和癌症进展。因此，药理学 特定压电同系物的调节可以帮助治疗许多人类疾病。然而，这一努力是 目前受到缺乏同源选择性药理学试剂和缺乏 清楚的分子理解，压电通道打开和关闭其孔， 物理和化学刺激。为了弥补这一差距，该提案将利用最近确定的 哺乳动物Piezo 1通道中的药理学结合位点。具体来说，我们将使用反向 遗传学、高分辨率电生理学和钙成像来解剖结构通路 由此小分子Yoda 1的结合有力地促进开放状态（Aim 1）。 与此同时，我们将部署一种创新的，多管齐下的方法， 计算，结构-活性关系，基于机器学习的超- 大的十亿化合物库，以及快速实验筛选分析，以确定新的 优先结合导电或非导电通道构象的分子，因此 可能分别作为Piezo 1激活剂和抑制剂（目的2）。成功完成 该项目的一部分将确定与以下相关的离散机械传导途径： Piezo 1通道的药理学激活，并鉴定新的药理学激活剂， Piezo 1通道抑制剂，具有潜在的研究和临床价值。

Summary Mechanosensitive Piezo ion channels enable eukaryotic cells to sense mechanical forces. In vertebrates, two Piezo homologs, Piezo1 and Piezo2, play central roles in all major physiological systems and are associated with numerous diseases including hypertension, xerocytosis, lymphedema, arthrogryposis, inflammation, pain, and cancer progression. Hence, pharmacological modulation of specific Piezo homologs could help treat many human ailments. Yet, this effort is currently limited by the paucity of homolog-selective pharmacological agents and the lack of a clear molecular understanding by which Piezo channels open and close their pore in response to physical and chemical stimuli. To bridge this gap, this proposal will leverage a recently identified pharmacological binding site in mammalian Piezo1 channels. Specifically, we will use reverse genetics, high-resolution electrophysiology, and calcium imaging to dissect structural pathways by which the binding of the small molecule Yoda1 energetically promotes an open state (Aim 1). In parallel, we will deploy an innovative, multi-pronged approach combining binding free energy calculations, structure-activity relationships, machine learning-based virtual screening of ultra- large billion-compounds library, and rapid experimental screening assays to identify novel molecules that preferentially bind conducting or non-conducting channel conformations, thus potentially acting as Piezo1 activators and inhibitors, respectively (Aim 2). Successful completion of this project will identify discrete mechanotrasnduction pathways associated with pharmacological activation of Piezo1 channels and identify novel pharmacological activators and inhibitors of Piezo1 channels with potential research and clinical value.