Structural insights into small molecule activation of TRPC4/5 channels

细胞是所有已知生物体的基本单位，人类由许多不同类型的细胞组成，其中大多数是高度专业化的。为了正常工作，细胞需要与周围环境和相邻细胞进行通信。这需要信息穿过细胞膜传递，细胞膜将细胞内容物与细胞外环境分开。一个主要的通讯机制是离子-主要是钠，钾，钙和氯-通过位于膜内的通道形成蛋白质，所谓的离子通道穿过细胞膜的运动。许多人类疾病是由离子通道功能异常引起的，许多成功的治疗药物通过激活或阻断离子通道起作用。我们的研究重点是称为TRPC 4和TRPC 5通道的离子通道，这些通道越来越多地被认为是各种疾病的潜在药物靶点-包括癌症，心力衰竭，心血管和代谢疾病，癫痫和焦虑症-但对于这些疾病，激活剂和阻断剂的开发已被证明是困难的。例如，我们以前发现，从传统医学中使用的非洲树木中分离出的天然产物Englerin A通过有效激活TRPC 4通道选择性地杀死肾癌细胞。Englerin A也是TRPC 5通道的非常有效的激活剂。然而，Englerin A太不稳定，毒性太大，不能用作抗癌药物。在这个项目中，我们将研究Englerin A如何与TRPC 4和TRPC 5通道相互作用。我们将结合不同团队成员的具体专业知识，使用各种实验方法。例如，我们将使用可以与TRPC 4/5通道发生化学反应的Englerin A类似物，并使用质谱法来确定反应在通道中发生的位置。此外，我们将使用最先进的电子显微镜-利兹大学和威康信托基金会最近投资1700万英镑的一部分-来确定TRPC 4/5通道及其与Englerin A的复合物的三维结构。这些结果将揭示Englerin A如何在分子水平上工作，以及TRPC 4/5通道的活性如何被小分子调节。这将使未来能够开发靶向特定TRPC 4或TRPC 5通道的药物，这可能导致靶向这些通道的第一种药物的开发。我们将通过与马克斯·普朗克学会的领先发现中心的持续合作，确保未来在药物发现过程中使用我们的结果，旨在开发用于临床试验的药物样分子。此外，我们将在开放获取出版物中发布我们的结果，并通过公共存储库免费提供我们的数据和材料。

The cell is the basic unit of all known living organisms, and humans consist of many different types of cells, the majority of which are highly specialised. In order to function properly, cells need to communicate with their environment and with neighbouring cells. This requires the transmission of information across the cell membrane, which separates the cellular content from the extracellular environment. One major mechanism of communication is the movement across cell membranes of ions - mainly sodium, potassium, calcium and chloride - through channel-forming proteins that are located within the membrane, so-called ion channels. Many human diseases result from abnormalities in the function of ion channels, and many successful therapeutic drugs work by activating or blocking ion channels. Our research focuses on ion channels called TRPC4 and TRPC5 channels, which are increasingly recognised as potential drug targets in a variety of diseases - including cancer, heart failure, cardiovascular and metabolic disease, epilepsy and anxiety disorders - but for which the development of activators and blockers as drugs has proven difficult. For example, we previously discovered that Englerin A, a natural product isolated from an African tree used in traditional medicine, selectively kills renal cancer cells by the potent activation of TRPC4 channels. Englerin A is also a very potent activator of TRPC5 channels. However, Englerin A is too unstable and too toxic to be used as an anti-cancer drug. In this project, we will study how Englerin A interacts with TRPC4 and TRPC5 channels . We will use a combination of experimental approaches, building on the specific expertise of the different team members. For example, we will use analogues of Englerin A that can chemically react with TRPC4/5 channels, and use mass spectrometry to identify where in the channels the reactions take place. In addition, we will use state-of-the-art electron microscopes - part of a recent £17m investment by the University of Leeds and the Wellcome Trust - to determine the three-dimensional structures of TRPC4/5 channels and their complexes with Englerin A.These results will reveal how Englerin A works on the molecular level, and how the activity of TRPC4/5 channels can be regulated by small molecules. This will enable future development of drugs that targets specific TRPC4 or TRPC5 channels, which may lead to the development of the first drugs that target these channels. We will ensure the future use of our results in the drug discovery process through our ongoing collaboration with the Lead Discovery Center of the Max Planck Society, with the aim to develop drug-like molecules for clinical trials. In addition, we will publish our results in open access publications, and make our data and materials freely available through public repositories.