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

• 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
• 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

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 败血症是美国第十大死因。尚未解决的系统性细胞因子风暴 由细菌感染引起是败血症的标志。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)，触发有害的 影响包括多器官衰竭。尽管进行了广泛的研究，脓毒症的治疗方法仍集中在 使用导致多重耐药性的抗菌药物。因此，一个未满足的科学需求是 了解抗炎反应的分子调节，从而减轻炎症的严重程度 组织损伤。单一免疫球蛋白白细胞介素 1 相关受体 (SIGIRR)，也是 称为 Toll/IL-1 受体 8，对 TLR 和 ILR 具有抗炎作用 发信号。最近，IL-37 被认为是先天免疫的抑制因子。 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。受体降解是受体功能的负反馈调节，是一种 通过翻译后修饰（例如磷酸化和 泛素化。泛素化是蛋白质降解的分子信号 蛋白酶体或溶酶体。去泛素化，由去泛素化酶介导 （DUB），通过从目标蛋白中去除泛素链来严格控制蛋白稳定性。在 根据我们的初步数据，我们发现 (i) SIGIRR 在 蛋白酶体对其配体结合做出反应； (ii) 泛素特异性蛋白酶 (USP13) DUB 成员，通过水解泛素链来靶向并稳定 SIGIRR 信号灯； (iii) 糖原合成酶激酶 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； (iv) USP13 提高实验性脓毒症的存活率。这些观察导致了以下结果 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。为了评估这个假设，我们将 确定 1) USP13 促进的 SIGIRR 稳定性的分子特征； 2) 如果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。在 总之，该应用提供了 SIGIRR 降解的分子机制 磷酸化驱动的泛素化。我们的初步数据此前已发现了两个 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，它de- 泛素化 SIGIRR。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 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。