Mechanism of Muscimol as a Novel Pyroptosis Inhibitor

蝇蕈醇作为新型焦亡抑制剂的作用机制

• 批准号: 10724728
• 负责人: Susan Leilani Fink
• 金额: $ 19.44万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724728
• 关键词: Affect; Agonist; Area; Binding Proteins; CASP1 gene; Caspase; Cell Death; Cell Death Process; Cell membrane; Cells; Coupled; Cytolysis; Data; Development; Disease; Exposure to; Family; Foundations; Future; GABA Agonists; GABA Receptor; Glycine; Goals; Immune signaling; Inflammasome; Inflammatory; Intervention; Lytic; Mediating; Modeling; Molecular; Muscimol; Myocardial Infarction; Nervous System Physiology; Neurons; Parents; Pathogenesis; Pathology; Peptide Hydrolases; Peptides; Process; Property; Proteins; Proteomics; Research Project Grants; Role; Rupture; Sepsis; Shotguns; Site; Stimulus; Stroke; Structure-Activity Relationship; System; Testing; Therapeutic; Ultraviolet Rays; VDAC1 gene; analog; chemical property; experimental study; human disease; in vivo evaluation; inhibitor; innovation; member; mortality; novel; novel therapeutic intervention; prevent; programs; receptor; release factor; small molecule; systemic inflammatory response; therapeutic target

Project Summary Pyroptosis is a programmed process of lytic, pro-inflammatory cell death that is involved in the pathogenesis of leading global causes of mortality. Inflammasome-mediated innate immune signaling activates caspase-1 family proteases to initiate pyroptosis by cleaving the pore-forming protein, gasdermin D. Recent data demonstrate that the protein ninjurin-1 oligomerizes during pyroptosis and is required for plasma membrane rupture, or cell lysis, downstream of gasdermin D pore formation. Cellular factors released during pyroptotic lysis cause local and systemic inflammation and pathology, but processes that regulate plasma membrane rupture and whether these can be therapeutically targeted, are not well-understood. We recently identified muscimol as a novel inhibitor of pyroptotic lysis, but its mechanism of action is not yet known. Muscimol is well-studied as an agonist of neuronal GABA receptors, but our preliminary data suggest that inhibition of pyroptotic lysis is not mediated by these receptors. This proposal aims to understand how muscimol prevents pyroptotic lysis and identify muscimol analogs with potent and specific activity. The experiments outlined in this proposal will systematically examine steps in the process of pyroptosis for inhibition by muscimol. Based on preliminary data, we will focus experiments on the hypothesis that muscimol interferes with ninjurin-1 oligomerization, while also testing other possibilities. We will use complementary models of pyroptosis induced by inflammasome-dependent and -independent stimuli, and further employ reductionist systems based on our findings. Our preliminary data suggest that there are specific molecular determinants for muscimol inhibition of pyroptotic lysis, as analogs demonstrate varied potency from the parent molecule, not correlating with GABA receptor activity. We will systematically test a panel of rationally-selected, already synthesized, muscimol analogs for inhibition of pyroptotic lysis. We hypothesize that our results will reveal a novel structure-activity relationship for muscimol inhibition of plasma membrane rupture compared to its canonical activity at neuronal receptors. In addition, these experiments may yield analogs with increased potency and / or specific activity to prevent pyroptotic lysis, without activity at GABA receptors. Finally, we will utilize the unique chemical properties of muscimol, coupled with advances in proteomics and the expertise of our collaborators, to identify novel muscimol-binding proteins. Together, the results of these experiments will inform a precise molecular understanding of the mechanism of action to disrupt pyroptotic lysis and provide the foundation for a novel therapeutic strategy for the many diseases in which pyroptosis has been implicated.

项目摘要 凋亡是裂解的促炎细胞死亡的编程过程，与 主要的全球死亡原因。炎性介导的先天免疫信号传导激活caspase-1 通过裂解孔形成蛋白，GasderminD来启动凋亡的家族蛋白酶D 证明蛋白质忍者-1在凋亡过程中会寡聚，并且需要质膜 破裂或细胞裂解，在加油动物D孔的下游。凋亡期间释放的细胞因子 裂解会导致局部和全身性炎症和病理学，但调节质膜的过程 破裂以及是否可以将其定位为靶向，这不是很好地理解。我们最近确定了 麝香酚是一种新型的凋亡裂解抑制剂，但其作用机理尚不清楚。 Muscimol是 作为神经元GABA受体的激动剂的研究充分，但是我们的初步数据表明抑制 凋亡裂解不是由这些受体介导的。该建议旨在了解麝香酚如何阻止 凋亡裂解并鉴定具有有效和特定活性的麝香酚类似物。 该提案中概述的实验将系统地检查在凋亡过程中的步骤 麝香酚的抑制作用。根据初步数据，我们将重点进行实验，以下假设 干扰忍者-1寡聚化，同时还测试其他可能性。我们将使用补充 由炎症体依赖性和非依赖性刺激引起的凋亡模型，并进一步采用 基于我们的发现，还原主义系统。我们的初步数据表明有特定的分子 肌酚抑制凋亡裂解的决定因素，因为类似物表明父母的效力各不相同 分子，与GABA受体活性无关。我们将系统地测试一个理性选择的面板， 已经合成的肌菌酚类似物以抑制凋亡裂解。我们假设我们的结果将会 揭示了肌菌酚抑制质膜破裂的新型结构活性关系 它在神经元受体上的规范活性。另外，这些实验可能会产生类似物随着增加而产生的 效力和 /或特定活性可预防凋亡裂解，而在GABA受体上没有活性。最后，我们会的 利用麝香酚的独特化学特性，再加上蛋白质组学的进步和专业知识 我们的合作者，以识别新型的麝香酚结合蛋白。这些实验的结果在一起将 告知对破坏凋亡裂解的作用机理的精确分子理解，并提供 为涉及凋亡的许多疾病的新型治疗策略的基础。