Necroinflammatory Cell Death in Sepsis

项目摘要/摘要： 脓毒症是一种致命的感染，其特征是宿主免疫反应失调。结果却没能 尽管进行了几十年的研究，但仍在不断改进。败血症的免疫反应是多种多样的。免疫疗法失败了 部分原因是该综合征的异质性。除了免疫沉默的细胞凋亡外， 坏死性炎症细胞死亡，通常是坏死性上睑下垂，具有免疫刺激性，可持续存在。 脓毒症中的炎症。坏死炎性细胞死亡的启动和协调是复杂的。肿瘤坏死因子相关 凋亡诱导配体(TRAIL)协调与细胞凋亡增加相关的细胞过程 坏死性炎症细胞死亡。受体相互作用丝氨酸/苏氨酸激酶3(RIPK3)在坏死性上皮病中的作用 细胞死亡。我们的工作表明在重症监护病房的脓毒症患者中RIPK3平行增加 器官功能障碍加重，并与不良结局相关。在重症监护室，我们已经证明了 较低的TRAIL与较高的RIPK3和增加的器官功能障碍相关。在这个项目中，我们将检查 TRAIL和RIPK3在三个时间点，在急诊科和ICU。我们假设 以高RIPK3和低TRAIL为特征的坏死性炎症细胞死亡将识别进展为 脓毒症和感染性休克，患者将出现新的坏死性炎症细胞死亡模式。 脓毒症死亡风险增加。目标1将在三个关键时间点创建一个人类患者队列 在医院的一次急性入院期间。第一次是在急诊科入院后不久 复苏和抗菌治疗的管理。后续的抽血结果如下 当器官功能障碍确定且治疗已开始时，进入病房或ICU。目标2将 通过两种途径研究TRAIL和RIPK3与败血症、感染性休克和死亡率的关系 方法论。第一项研究将检查TRAIL和RIPK3是否将提高我们诊断脓毒症的能力 结合急诊科的生理性脓毒症预测工具。第二个将评估 在对时间依赖性患者的影响建模后，随访TRAIL和RIPK3对预后的影响， 病原菌和治疗因素。对于AIM 3，我们将测量定向机械性细胞死亡小组的水平 包括RIPK1、RIPK3、MLKL，以及与损伤相关的关键分子模式、线粒体DNA和HMGB1。 我们还将评估蛋白质组学平台中更广泛的坏死性炎症生物标记物小组。到时候我们会的 评估是否存在由相对生物标记物水平和生理指标定义的患者群 变量。我们将检查这些患者群是否具有不同的结果。如果这项提案的目的是 ，我们将获得有关坏死性炎症细胞死亡在人类脓毒症中的作用的有用信息。 从多个时间点。这项研究的结果可能为研究生物标记物的发展提供洞察力 针对坏死性炎症细胞死亡的临床试验的预测性丰富。

PROJECT SUMMARY/ABSTRACT: Sepsis is a deadly infection characterized by a dysregulated host immune response. Outcomes have failed to improve despite decades of research. The immune response in sepsis is varied. Immunologic therapy has failed in part due to the heterogeneity of the syndrome. Beyond the immunologically silent apoptosis, necroinflammatory cell death, commonly necroptosis, is immunologically stimulating and can perpetuate inflammation in sepsis. The initiation and coordination of necroinflammatory cell death is complex. TNF related apoptosis inducing ligand (TRAIL) coordinates cellular processes associated with increased apoptosis and necroinflammatory cell death. Receptor interacting serine/threonine kinase 3 (RIPK3) is essential to necroptotic cell death. Our work has shown that RIPK3 is increased in septic patients in the intensive care unit in parallel with increased organ dysfunction and is associated with poor outcomes. In the ICU, we have demonstrated that lower TRAIL is associated with higher RIPK3 and increased organ dysfunction. In this project, we will examine TRAIL and RIPK3 at three time points, in the emergency department and ICU. We hypothesize that necroinflammatory cell death, characterized by high RIPK3 and low TRAIL will identify those who progress to sepsis and septic shock and that there will be novel patterns of necroinflammatory cell death in patients at increased risk of death with sepsis. AIM 1 will create a human cohort of patients at three critical time points during an acute admission to the hospital. The first is soon after admission to the emergency department prior to resuscitation and the administration of antimicrobial therapy. The follow up blood draws are obtained following admission to the ward or ICU when organ dysfunction is established and therapy has been initiated. AIM 2 will examine the relationship between TRAIL and RIPK3 and sepsis, septic shock and mortality through two methodologies. The first will examine whether TRAIL and RIPK3 will increase our ability to diagnose sepsis when combined with physiologic sepsis prediction tools in the emergency department. The second will evaluate the effect of the follow up TRAIL and RIPK3 on outcomes, after modeling the effect of time dependent patient, pathogen and treatment factors. For AIM 3, we will measure levels of a targeted mechanistic cell death panel including, RIPK1, RIPK3, MLKL, along with key damage associated molecular patterns, mtDNA and HMGB1. We will also evaluate a broader necroinflammatory biomarker panel in a proteomics platform. We will then evaluate whether there are clusters of patients defined by relative biomarker levels together with physiologic variables. We will examine if these patient clusters have differential outcomes. If the aims of this proposal are achieved, we will have useful information concerning the role of necroinflammatory cell death in human sepsis from multiple time points. Results from this study may offer insight into the development of biomarkers for predictive enrichment of clinical trials targeting necroinflammatory cell death.