Molecular regulation of anti-inflammatory cytokine receptor in sepsis

脓毒症抗炎细胞因子受体的分子调控

基本信息

批准号：
9912821
负责人：
Yutong Zhao
金额：
$ 42.5万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-11-10 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9912821
关键词：
Acute Acute Lung Injury Anti-Inflammatory Agents Antiinflammatory Effect Bacterial Infections Cause of Death Cells Cytokine Receptors Data Deubiquitinating Enzyme Deubiquitination Docking Endotoxins Etiology Excision Exhibits Failure Feedback Foundations Gram-Negative Bacterial Infections Immunoglobulins Impairment Infection Inflammation Inflammatory Inflammatory Response Interleukin-1 Interleukin-1 Receptors Interleukins Interruption Lead Ligand Binding Ligands Link Lung Lysosomes Maintenance Mediating Mediator of activation protein Molecular Molecular Profiling Morbidity - disease rate Multi-Drug Resistance Natural Immunity Pathogenesis Phosphorylation Polyubiquitination Post-Translational Protein Processing Process Protein Kinase Proteins Receptor Signaling Regulation Research Role Scheme Sepsis Septic Shock Severities Signaling Protein Site Survival Rate Tissues Toll-like receptors Ubiquitin Ubiquitination Up-Regulation Virulent antimicrobial cytokine release syndrome glycogen synthase kinase 3 beta knock-down lung injury member mortality multicatalytic endopeptidase complex novel therapeutics pathogen protein degradation receptor receptor function response septic therapeutic target tissue injury ubiquitin-specific protease

项目摘要

Abstract Sepsis is the 10th leading cause of death in the US. An unresolved systemic cytokine storm caused by bacterial infection is a hallmark of sepsis. The robust acute inflammatory response, through Toll Like Receptors (TLRs) and interleukin-1R like receptors (ILRs), trigger detrimental effects including multi-organ failure. Despite extensive research, therapies for sepsis focus on the use of antimicrobials that lead to multi-drug resistance. Hence, an unmet scientific need is to understand the molecular regulation of anti-inflammatory responses that diminish the severity of tissue injury. Single immunoglobulin interleukin-1-related receptor (SIGIRR), which is also known as Toll/IL-1 receptor 8, exhibits an anti-inflammatory effect against TLRs and ILRs signaling. Recently, IL-37, which is a suppressor of innate immunity, has been identified as the SIGIRR ligand. Both IL-37 and SIGIRR have been recognized as major therapeutic targets to lessen cytokine storm, however, very little is known regarding the molecular regulation of SIGIRR stability. Receptor degradation, a negative feedback regulation of receptor function, is a highly regulated process by post-translational modification, such as phosphorylation and ubiquitination. Ubiquitination is a molecular signal for protein degradation in either the proteasome or lysosome. De-ubiquitination, which is mediated by deubiquitinating enzymes (DUBs), tightly controls protein stability by removal of ubiquitin chains from target proteins. In our preliminary data, we discovered that (i) SIGIRR is poly-ubiquitinated and degraded in the proteasome in response to its ligand binding; (ii) Ubiquitin-specific protease (USP13), a member of DUBs, targets and stabilizes SIGIRR by hydrolyzing the ubiquitin chains from SIGIRR; (iii) glycogen synthase kinase 3β (GSK3β) phosphorylates SIGIRR and interrupts the association between SIGIRR and USP13, thereby reducing SIGIRR stability; (iv) USP13 increases survival rate in experimental sepsis. These observations have led to the following hypothesis: USP13 ameliorates cytokine storm and septic shock through deubiquitination and stabilization of the anti-inflammatory receptor, SIGIRR. To evaluate this hypothesis we will determine 1) molecular signature of USP13-promoted SIGIRR stability; 2) if GSK3β-induced disruption of USP13/SIGIRR interaction lessens anti-inflammatory effects of SIGIRR; 3) if stabilization of SIGIRR by USP13 mitigates endotoxin-induced pro-inflammatory responses. In summary, this application provides molecular mechanisms by which SIGIRR is degraded via phosphorylation-driven ubiquitination. Our preliminary data has uncovered two previously unrecognized post-translational modifications of SIGIRR: phosphorylation and ubiquitination. Two mediators were revealed: GSK3β, which phosphorylates SIGIRR; and USP13, which de- ubiquitinates SIGIRR. These studies will be the first to elucidate that phosphorylation of SIGIRR promotes its ubiquitination by disassociating USP13 from SIGIRR. These studies will lay the foundation for a significant mechanistic advance regarding the molecular regulation of the inflammatory response through modulation of anti-inflammatory receptor stability.

抽象的脓毒症是美国第十大死因，是一场尚未解决的系统性细胞因子风暴。由细菌感染引起的脓毒症的一个标志是强烈的急性炎症反应。通过 Toll 样受体 (TLR) 和白细胞介素 1R 样受体 (ILR)，引发痛苦尽管进行了广泛的研究，脓毒症的治疗仍集中在包括多器官衰竭在内的影响上。因此，未满足的科学需求是使用导致多重耐药性的抗菌药物。了解抗炎反应的分子调节，从而减轻炎症的严重程度单一免疫球蛋白白细胞介素 1 相关受体 (SIGIRR)，这也是组织损伤。称为 Toll/IL-1 受体 8，对 TLR 和 ILR 具有抗炎作用最近，IL-37（一种先天免疫抑制剂）已被确定为 SIGIRR 配体。IL-37 和 SIGIRR 均已被认为是主要的治疗靶点。减少细胞因子风暴，然而，关于细胞因子的分子调节知之甚少 SIGIRR 稳定性是受体功能的负反馈调节。通过翻译后修饰（例如磷酸化和泛素化是蛋白质降解的分子信号。蛋白酶体或溶酶体，由去泛素化酶介导。（DUB），通过从目标蛋白质中去除泛素链来严格控制蛋白质稳定性。我们的初步数据发现，(i) SIGIRR 是多泛素化的，并且在蛋白酶体对其配体结合作出反应；(ii) 泛素特异性蛋白酶 (USP13)，一种 DUB 成员，通过水解泛素链来靶向并稳定 SIGIRR SIGIRR；(iii) 糖原合酶激酶 3β (GSK3β) 磷酸化 SIGIRR 并中断 SIGIRR 和 USP13 之间的关联，从而降低 SIGIRR 稳定性；增加实验性脓毒症的存活率这些观察结果导致以下结果。假设：USP13 通过去泛素化和改善细胞因子风暴和败血性休克抗炎受体 SIGIRR 的稳定性为了评估这一假设，我们将进行评估。确定 1) USP13 促进的 SIGIRR 稳定性的分子特征 2) 是否由 GSK3β 诱导； USP13/SIGIRR 相互作用的破坏会降低 SIGIRR 的抗炎作用 3) 如果 USP13 稳定 SIGIRR 可减轻内毒素诱导的促炎症反应。总之，该应用提供了 SIGIRR 降解的分子机制我们的初步数据之前发现了两个磷酸化驱动的泛素化。 SIGIRR 未被识别的翻译后修饰：磷酸化和泛素化。揭示了两种介质：GSK3β（磷酸化 SIGIRR）和 USP13（去磷酸化）；这些研究将首次阐明 SIGIRR 的磷酸化。通过将 USP13 与 SIGIRR 分离来促进其泛素化。这些研究将奠定基础。为分子调控方面的重大机械进展奠定了基础通过调节抗炎受体稳定性来调节炎症反应。