The Inflammasome: A Novel Biomarker in ALI/ARDS

描述（由申请人提供）：炎性小体最近被描述为一种重要的蛋白质复合物，其中非nfkb介导的信号通路导致关键的促炎细胞因子上调，包括白细胞介素(IL)- 1b和IL-18。我们小组的成员最近报道了炎症小体在小鼠脓毒症的前炎症反应中起关键作用，这种反应依赖于线粒体完整性和对损伤的完整自噬反应。我们现在提供的数据支持炎症小体在预测人类感染相关ALI/ARDS的严重程度和死亡率方面的重要作用。此外，我们的初步动物研究表明，他汀类药物通过激活炎性体加重肺损伤和炎症。因此，我们假设炎症小体的激活在感染相关ALI/ARDS的发展中起着关键作用，并且他汀类药物可能会增加肺损伤期间炎症小体相关的下游细胞因子。特别值得一提的是，我们与ARDSnet SAILS试验研究人员（他汀类药物与安慰剂在感染相关ALI/ARDS中的随机试验）在临床试验RFA （HL-12-012）中的合作提供了一个独特的机会来获得额外的关键人类样本收集，以解决这些重要的过程。因此，我们提出：具体目标1：通过前瞻性收集安慰剂和他汀类药物治疗的风帆受试者的血液（n=100）和血浆样本（n=600），确定感染相关ALI/ARDS期间炎症体的基因表达和蛋白质水平。炎症小体的基因表达和蛋白水平将与60天死亡率和其他SAILS试验次要结果相关。我们假设循环炎症小体水平将作为感染相关ALI/ARDS严重程度和死亡率的生物标志物，并且他汀类药物治疗的受试者的炎症小体水平将与临床结果相关。特异性目的2：确定炎性小体复合物表达的细胞定位以及炎性小体激活对细胞反应和功能的作用，使用从前瞻性招募的安慰剂和他汀类药物治疗的sail受试者（n=100）中分离的原代中性粒细胞和单核细胞，以及从对照ICU受试者（n=100）分离的原代细胞。我们假设，确定哪些循环细胞是炎性小体激活的主要来源将增加炎性小体水平与临床结果相关性的敏感性，并且炎性小体的局部激活将导致线粒体功能障碍，这将在他汀类药物的存在下增强。

DESCRIPTION (provided by applicant): The inflammasome has recently been described as an important protein complex in which a non-NFkB- mediated signaling pathway leads to up-regulation of key pro-inflammatory cytokines, including interleukin (IL)- 1b and IL-18. Members of our group recently reported that the inflammasome plays a critical role in pro- inflammatory response in murine sepsis, and this response is dependent upon mitochondrial integrity and an intact autophagy response to injury. We now present data supporting an important role for the inflammasome in predicting severity and mortality during human infection-related ALI/ARDS. Moreover, our preliminary animal studies demonstrate that statins exacerbate lung injury and inflammation via activation of the inflammasome. We therefore hypothesize that activation of the inflammasome plays a critical role in the development of infection-related ALI/ARDS and that statin administration may increase inflammasome-related downstream cytokines during lung injury. In particular, our collaboration with the ARDSnet SAILS trial investigators (a randomized trial of statins vs. placebo in infection-related ALI/ARDS) in this Ancillary Studies in Clinical Trials RFA (HL-12-012) provides a unique opportunity to obtain additional collection of key human samples to address these important processes. We therefore propose: Specific Aim 1: To determine gene expression and protein levels of the inflammasome during infection-related ALI/ARDS using prospectively collected blood (n=100) and banked plasma samples (n=600) from placebo- and statin-treated SAILS subjects. Gene expression and protein levels of the inflammasome will be correlated with 60-day mortality and additional SAILS trial secondary outcomes. We hypothesize that circulating inflammasome levels will serve as a biomarker of severity and mortality of infection-related ALI/ARDS and that inflammasome levels in statin-treated subjects will correlate with clinical outcomes. Specific Aim 2: To determine the cellular localization of expression of the inflammasome complex and role of inflammasome activation on cellular responses and function, using primary neutrophils and monocytes isolated from prospectively enrolled placebo- and statin-treated SAILS subjects (n=100), as well as primary cells isolated from control ICU subjects (n=100). We hypothesize that determining which circulating cells are the predominant source of inflammasome activation will increase sensitivity of the correlation of inflammasome levels with clinical outcomes and that localized activation of the inflammasome will result in mitochondrial dysfunction that will be enhanced in the presence of statin administration.