Pannexin channels in tissue inflammation and metabolite release

Pannexin 参与组织炎症和代谢物释放的通道

• 批准号: 9894839
• 负责人: Kodi S Ravichandran
• 金额: $ 39.31万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9894839
• 关键词: Address; Adenosine; Agonist; Airway Disease; Anti-Inflammatory Agents; Apoptosis; Apoptotic; Binding; Blood Pressure; Blood Vessels; CD4 Positive T Lymphocytes; Cardiovascular Pathology; Cardiovascular system; Caspase; Cell Communication; Cell Survival; Cell membrane; Cells; Collaborations; Communication; Cues; Disease; Environment; Eragrostis; Excision; Generations; Goals; Hypertension; Inflammation; Inflammatory; Ions; Laboratories; Link; Lung Inflammation; Mediating; Mediator of activation protein; Modeling; Mouse Strains; Mus; Nature; Nucleotides; Pathologic; Phagocytes; Phosphotransferases; Physiological; Physiological Processes; Physiology; Process; Regulatory T-Lymphocyte; Research; Respiratory System; Role; Signal Transduction; Spironolactone; T-Lymphocyte; Testing; Time; Tissues; Transgenic Mice; Work; airway inflammation; base; blood pressure regulation; cell type; effector T cell; in vivo; inhibitor/antagonist; insight; intercellular communication; member; metabolomics; methoctramine; mouse model; novel; overexpression; programs; protein phosphatase inhibitor-2; salt-inducible kinase; small molecule; synergism; tool; transcriptomics; trovafloxacin; vascular inflammation; yeast two hybrid system

PROJECT 1 PROJECT SUMMARY Communication between cells within a tissue environment is fundamentally important for many physiological processes. Working in the context of how apoptotic cells are removed by phagocytes within tissues, we initially observed that nucleotides such as ATP and UTP released from apoptotic cells via Pannexin-1 (Panx1) channels and act as ‘find-me signals’ to attract phagocytes, leading to the prompt removal of corpses. Subsequent work (in collaboration with the other three Project leaders) identified a caspase-dependent cleavage of the C-terminus of Panx1 as one of the mechanisms of Panx1 channel opening. In the past few years, directly via Project 1, we have made several key observations that form the basis of this current renewal. These include the identification of a new Panx1 channel inhibitor trovafloxacin and demonstrating a novel role for Panx1 in apoptosis (Poon et al., Nature 2014); identifying spironolactone as a new inhibitor of Panx1 and demonstrating the importance spironolactone mediated Panx1 inhibition in regulating blood pressure spironolactone (Good et al., Circ Research 2017); identified a new activator of Panx1; generating a new transgenic mouse capable of inducibly overexpressing Panx1; and, identification of new binding partners of Panx1 (kinase SIK) via a two-hybrid screen. Project 1 has also generated a number of new mouse tools that are widely used by other members of the PPG. In this Project 1, we aim to test the central hypothesis that Panx1 controls inflammatory processes through purinergic as well as non-purinergic signals, fundamentally shifting the current perspective on Panx1. In Aim 1, we will test a new concept (from our preliminary studies) that Panx1 channels can release both purinergic and non-purinergic metabolites that influence inter-cellular communication directly, and the anti-inflammatory tone within the local tissue milieu. We will also test the role of a newly identified Panx1 agonist methoctramine, and whether this can also influence the release of novel metabolites via Panx1. In Aim 2, based on our preliminary studies that Panx1 channels are required for limiting airway inflammation, we will test the role of Panx1 in airway disease using models we have recently established (Han et al., Nature 2016). Further, based on the new Panx1 interacting partner SIK that we identified, we will probe the mechanistic aspects of Panx1 requirement in airway inflammation. Collectively, we expect these studies to provide new insights on how pannexin channels function in different contexts in vivo, identify new modes of stimulating Panx1 function, and better define Panx1- dependent intercellular communication toward regulating the inflammatory tone in tissues. The new mouse lines, small molecule modulators, and protein interactors identified by Project 1 will integrate seamlessly with the other three projects and cores within this P01.

项目1项目摘要 组织环境中细胞之间的通信对于许多生理学对于许多生理来说至关重要 过程。在组织中如何通过吞噬细胞去除凋亡细胞的背景下工作，我们最初 观察到通过pannexin-1（Panx1）通道从凋亡细胞释放的ATP和UTP等核动肽 并充当吸引吞噬细胞的“发现信号”，从而迅速去除尸体。随后的工作 （与其他三个项目负责人合作）确定了C-terminus的caspase依赖性裂解 panx1是panx1通道开放的机制之一。在过去的几年中，直接通过项目1，我们 已经进行了几个关键观察，构成了当前续约的基础。这些包括身份证明 新的PANX1通道抑制剂trovafloxacin，并证明了Panx1在凋亡中的新作用（POON ET） Al。，自然2014）;识别螺内酯作为PANX1的新抑制剂并证明进口 螺内酯介导的PANX1抑制在控制血压螺内酯（Good等，Circ Reseind 2017）;确定了panx1的新激活剂；产生一种能够诱导的新的转基因小鼠 过表达panx1;并且，通过两个杂交屏幕鉴定Panx1（激酶SIK）的新结合伙伴。 项目1还生成了许多新的鼠标工具，这些工具被PPG的其他成员广泛使用。 在该项目1中，我们旨在测试PANX1通过 嘌呤能以及非核能信号从根本上转移了PANX1的当前视角。在AIM 1中， 我们将测试一个新概念（从我们的初步研究中），PANX1通道可以释放嘌呤能和 直接影响细胞间交流和抗炎张力的非核能代谢产物 在当地的组织环境中。我们还将测试新鉴定的panx1激动剂甲胺的作用， 这是否也可以通过PANX1影响新代谢产物的释放。在AIM 2中，根据我们的初步 研究PANX1通道是限制气道注入所必需的，我们将测试PANX1在气道中的作用 我们最近建立了使用模型的疾病（Han等，自然，2016年）。此外，基于新的panx1 我们确定的互动合作伙伴SIK，我们将探测气道中PANX1需求的机械方面 炎。总的来说，我们希望这些研究能够提供有关Pannexin渠道如何功能的新见解 在体内不同的情况下，确定刺激PANX1功能的新模式，并更好地定义Panx1- 依赖的细胞间通信与组织中的炎症性张力。新的鼠标线， 小分子调节剂和项目1鉴定的蛋白质相互作用者将与另一个 该P01中的三个项目和核心。