Mechanisms of activation of lysosomal two-pore channel TPC2, a potential novel target for the treatment of lysosomal storage disorders and neurodegenerative diseases

溶酶体双孔通道TPC2的激活机制，治疗溶酶体贮积症和神经退行性疾病的潜在新靶点

• 批准号: 440563025
• 负责人: Professor Dr. Christian Michael Grimm
• 金额: --
• 依托单位: Walther-Straub-Institut für Pharmakologie und Toxikologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/440563025?language=en
• 关键词: Mechanisms; activation; lysosomal; two; pore

Endolysosomal ion channels, in particular the two-pore channels (TPCs) are currently garnering much interest due to its pathophysiological relevance (given links to several diseases) and from a structural standpoint (with emerging cryoEM structures). But uncertainty surrounding fundamental properties of TPCs including how they are activated and their ion permeability, coupled with a limited pharmacological tool set is slowing progress. This proposal will address these gaps in detail. We will 1) validate new chemical tools to study TPCs, in particular TPC2 in a physiological and pathophysiological context and we will 2) facilitate TPC2 drug discovery and design by laying the foundations for a clearer understanding of how TPC2 can conduct different ions in response to different activating cues with potentially differential physiological consequences. By using a combination of Ca2+ imaging, molecular biology and electrophysiology, we apply the first-in-kind lipophilic agonists of TPC2 that we have discovered to demonstrate that the ion selectivity of TPC2 is agonist-dependent and not fixed. This has very important implications for the physiological and pathophysiological roles and endogenous activation mechanisms of TPC2 and consequently for drug discovery and drug design. In order to establish TPC2 as a target to treat, e.g. LSDs or neurodegenerative diseases, insights into these different activation mechanisms and endogenous stimulation pathways are crucial and this work aims to deliver the missing knowledge gaps.

内溶酶体离子通道，特别是双孔通道（TPC）目前由于其病理生理学相关性（与几种疾病相关）和结构观点（具有新兴的cryoEM结构）而引起广泛关注。但是，围绕TPC基本特性的不确定性，包括它们如何被激活和它们的离子渗透性，加上有限的药理学工具集，正在减缓进展。本提案将详细处理这些差距。我们将1）验证新的化学工具来研究TPC，特别是在生理和病理生理背景下的TPC 2，我们将2）通过更清楚地理解TPC 2如何响应具有潜在差异生理后果的不同激活线索来传导不同离子，从而为TPC 2药物发现和设计奠定基础。通过使用Ca2+成像，分子生物学和电生理学的组合，我们应用我们发现的TPC 2的第一种亲脂性激动剂来证明TPC 2的离子选择性是激动剂依赖性的，并且不是固定的。这对TPC 2的生理和病理生理作用以及内源性激活机制以及药物发现和药物设计具有非常重要的意义。为了将TPC 2确定为治疗LSD或神经退行性疾病的靶点，对这些不同激活机制和内源性刺激途径的了解至关重要，这项工作旨在填补缺失的知识空白。