Inflammasome activation in trauma-hemorrhagic shock

项目摘要/摘要 控制炎症和细胞损伤是防治多器官衰竭的关键 (MOF)创伤后。然而，许多调节炎症和细胞死亡的机制 具体的器官仍不清楚。因此，尽管在多器官功能衰竭患者的支持性护理方面取得了进展，但 在多器官功能衰竭的治疗方面，或在我们充分预防多器官功能衰竭发病的能力方面，进展甚微。我们的首要目标是 最终为创伤患者开发基于调节炎症反应和 创伤和复苏失血性休克对细胞死亡的影响(HS/R)。在这份提案中，我们 将继续研究caspase-4(人)/11(鼠)对下游细胞和组织的特异性影响 我们还将评估一种新的caspase-4/11抑制剂降低HS/R的能力。 对全身炎症、细胞死亡和终末器官损伤的中介作用。了解多重影响 Caspase-11在炎症和器官损伤方面的研究将使我们能够评估caspase-11的翻译潜力 靶向抑制caspase-11可改善创伤和出血后患者的预后。 关于这一授权的最新工作定义了caspase-4/11激活的新机制(非 创伤/HS/R中的典型炎症体)Caspase-4/11是内毒素的细胞内受体 内毒素(LPs)，是感染和败血症过程中炎症反应的重要介质。我们的工作显示了 在非内毒素驱动的炎症模型中，我们定义了一种新的激活caspase-4/11的机制 Caspase-11由内源性氧化心磷脂(CLox)作用于线粒体外部。Clox是 受细胞色素c调节，细胞色素c的特异性抑制抑制caspase-11的激活，并高度 在HS/R小鼠模型中的器官保护作用，然而，caspase有害作用的机制基础是- 11在HS/R中的激活尚不清楚，可能是多因素的，因为其具有多细胞特异性炎症。 作用，包括诱导下垂(炎性细胞死亡)，主动释放炎性HMGB1 HS/R后肝细胞的胞外小泡(EV)及其对凝血的影响 通过增强组织因子(TF)与内皮细胞(EC)的结合，在脓毒症中发挥重要作用。因此，我们的主要 假设caspase-4/11在HS/R过程中通过多个细胞特异性激活是有害的 效果。在这项建议中，我们将：1：确定caspase-4/11介导的炎症在器官中的作用 HS/R时的损伤；2：确定caspase-4/11-激活对凝血和器官损伤的影响 在HS/R期间；3：在小鼠模型中确定caspase-11特异性抑制的安全性和有效性 我们期望在HS/R中显示caspase-4/11的特异性抑制是器官保护的，并且 保护机制是多因素的。我们还期望显示抑制caspase-4/11是一个有吸引力的 治疗目标是减少创伤引起的炎症、凝血障碍和器官损伤。

PROJECT SUMMARY/ABSTRACT Controlling inflammation and cellular damage is key to preventing and treating multiple organ failure (MOF) following trauma. However, many of the mechanisms that regulate inflammation and cell death in specific organs remain unknown. So despite advances in supportive care for MOF patients, there have been few advances in MOF treatments, or in our ability to adequately prevent MOF onset. Our overarching goal is to ultimately develop new therapeutics for trauma patients based on regulating inflammatory responses and effects on cell death following trauma and hemorrhagic shock with resuscitation (HS/R). In this proposal we will continue to investigate downstream cell and tissue-specific effects of caspase-4 (human)/11 (mouse) activation after HS/R. We will also assess the ability of a novel caspase-4/11 inhibitor to reduce HS/R- mediated effects on systemic inflammation, cell death and end-organ injury. Understanding the multiple effects of caspase-11 on inflammation and organ damage will enable us to assess the translational potential of targeted caspase-11-inhibition for improved patient outcomes after trauma and hemorrhage. The most recent work on this grant defined a novel mechanism of caspase-4/11 activation (non- canonical inflammasome) in trauma/HS/R. Caspase-4/11 is the intracellular receptor for lipopolysaccharide (LPS) and is an important mediator of inflammation during infection and sepsis. Our work showed activation of caspase-4/11 in non-LPS-driven models of inflammation, and we defined a novel mechanism of activation of caspase-11 by endogenous oxidized cardiolipin (CLox) on the outside of stressed mitochondria. CLox is regulated by cytochrome c, and specific inhibition of cytochrome c inhibits caspase-11 activation and is highly organ protective in a mouse model of HS/R. However, the mechanistic basis of detrimental effects of caspase- 11 activation in HS/R are not clear, and may be multifactorial given its multiple cell-specific inflammatory effects, including induction of pyroptosis (inflammatory cell death), active release of inflammatory HMGB1 in extracellular vesicles (EV) from hepatocytes after HS/R, and more recently-described effects on coagulation through enhanced binding of tissue factor (TF) on endothelial cells (EC) in sepsis. Therefore, our main hypothesis is that caspase-4/11 activation is detrimental during HS/R through multiple cell-specific effects. In this proposal we will: 1: determine the role of caspase-4/11-mediated inflammation on organ injury during HS/R; 2: determine the effects of caspase-4/11-activation on coagulation and organ injury during HS/R; 3: determine safety and effectiveness of caspase-11-specific inhibition in mouse models of HS/R. We expect to show specific inhibition of caspase-4/11 in HS/R is organ protective and the mechanisms of protection are multi-factorial. We also expect to show inhibition of caspase-4/11 is an attractive therapeutic target to reduce trauma-induced inflammation, coagulopathy and organ damage.