Neurosteroid Inhibition of Pyroptotic Lysis

神经类固醇抑制焦亡裂解

基本信息

批准号：
10171555
负责人：
Susan Leilani Fink
金额：
$ 23.33万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-25 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10171555
关键词：
Address Affect Allopregnanolone Biological CASP1 gene Cardiovascular Diseases Cell Death Cell Death Process Cell Volumes Cell membrane Cells Cleaved cell Clinical Chemistry Cytolysis Data Development Disease Event Experimental Designs Family Foundations Goals Inflammasome Intervention Ion Channel Libraries Lipid Binding Lipids Membrane Membrane Lipids Membrane Proteins Molecular Myocardial Infarction N-terminal Nerve Degeneration Nervous System Physiology Neurons Pathogenesis Pathology processes Peptide Hydrolases Physiological Positioning Attribute Process Proteins Regulation Role Sepsis Steroids Stroke Structure Structure-Activity Relationship Testing Therapeutic Vertebral column experimental study human disease inhibitor/antagonist innovation lipophilicity mortality nervous system disorder neurosteroids novel novel strategies pregnenolone sulfate prevent programs release factor repaired small molecule

项目摘要

Project Summary Pyroptosis is a program of cell death involved in the pathogenesis of leading global causes of mortality. Caspase-1 family proteases initiate pyroptosis by releasing the pore-forming portion of gasdermin D, which inserts into the plasma membrane leading to cell lysis. Cellular factors released during pyroptotic lysis cause local and systemic pathology, but processes regulating gasdermin D pore formation and lysis are not well understood. We recently identified the neurosteroid pregnenolone sulfate as a novel inhibitor of pyroptotic lysis, but its mechanism of action is not yet known. Neurosteroids including pregnenolone sulfate have a lipophilic backbone and interact with plasma membrane proteins and lipids, suggesting the hypothesis that these molecules may disrupt formation of gasdermin D pores. This proposal aims to understand how pregnenolone sulfate prevents pyroptotic lysis and determine whether other steroids share this activity. Our preliminary data demonstrate that pregnenolone sulfate prevents lysis during pyroptosis without affecting upstream activation of inflammasomes or caspase-1. The experiments outlined in this proposal will test the hypothesis that pregnenolone sulfate affects pore formation during pyroptosis and assess gasdermin D cleavage, plasma membrane localization and oligomerization. We will also address the alternative hypotheses that pregnenolone sulfate may regulate cell volume to prevent lysis or potentiate membrane repair processes. Together, the results of these experiments will inform a precise molecular understanding of the mechanism of action and reveal a novel strategy to disrupt pyroptotic lysis. The steroid backbone is shared by a large number of molecules, each with specific biological activities. Our preliminary data suggest that there may be molecular determinants for steroid inhibition of pyroptotic lysis, as a related steroid demonstrates reduced potency. We will test a rationally selected library of structurally distinct steroids for the ability to prevent pyroptotic lysis. These experiments will determine whether endogenous steroids demonstrate potency consistent with potential physiologic relevance. In addition, these results will identify the structure-activity relationship for inhibition of pyroptotic lysis to facilitate development of potent and specific molecules and define a new paradigm for disease intervention.

项目摘要凋亡是一个细胞死亡的程序，涉及主要全球死亡率的发病机理。 caspase-1家族蛋白酶通过释放加油的孔形成部分D启动凋亡插入导致细胞裂解的质膜中。凋亡过程中释放的细胞因子导致局部和系统的病理学，但是调节加油的过程的过程不好理解。我们最近确定神经类固醇硫酸盐是一种新型的凋亡抑制剂裂解，但其作用机理尚不清楚。包括硫酸妊娠烯醇酮在内的神经蛋白酶具有亲脂性主链并与质膜蛋白和脂质相互作用，表明假设是这些分子可能会破坏加油孔D孔的形成。该建议旨在了解如何孕妇硫酸盐可防止凋亡裂解，并确定其他类固醇是否共享这种活性。我们的初步数据表明，怀孕硫酸盐可防止凋亡过程中的裂解而不会影响炎症或caspase-1的上游激活。该提案中概述的实验将测试假设孕妇硫酸盐会影响凋亡过程中的孔形成并评估加油动物D 裂解，质膜定位和低聚。我们还将解决替代假设硫酸盐妊娠可以调节细胞体积以防止裂解或增强膜修复过程。总之，这些实验的结果将为对机制的机制提供精确的分子理解。动作并揭示了破坏凋亡裂解的新策略。类固醇主链由大量分子共享，每个分子都具有特定的生物学活性。我们的初步数据表明，可能存在类固醇抑制凋亡裂解的分子决定因素，作为一种相关的类固醇表现出降低的效力。我们将测试一个结构上不同的合理选择的库类固醇可预防凋亡裂解的能力。这些实验将确定内源性是否类固醇表现出与潜在的生理相关性一致的效力。此外，这些结果将确定抑制凋亡裂解的结构活性关系，以促进有效和特定的分子并定义了用于疾病干预的新范式。