Targeting DAMPs in Alcoholic Hepatitis

描述(申请人提供)：酒精性肝炎(AH)是酒精性肝病(ALD)的一种特别严重的形式，影响多达20%的酗酒者，具有高死亡率。然而，引发急性肝炎的分子途径仍不清楚，酒精性肝炎的治疗在过去30年中没有改变。值得注意的是，靶向炎性细胞因子，如肿瘤坏死因子！或减少ROS的产生都不成功，使用皮质类固醇的非特异性抗炎治疗只能起到适度的效果。在这项提案中，我们将 研究来自死亡细胞的标志性分子，称为损伤相关分子模式(DAMP)，有助于急性肝炎时肝脏和全身炎症，祖细胞增殖，并加重肝脏损伤和预后。这项拟议的研究将研究几种已知在肝脏无菌炎症和中性粒细胞募集中起作用的DAMP，特别关注(I)线粒体(MT)DAMP，因为它们在肝细胞中含量丰富，以及ALD中的线粒体变化，以及(Ii)HMGB1，坏死性肝损伤中的关键DAMP。我们的应用将结合对急性脑出血患者的人体研究和功能性小鼠研究，以确定与急性呼吸窘迫综合征高度相关的关键和可用药的湿通路。拟议的人体研究将与InTeam财团的项目1、人体生物库核心和10个临床中心合作进行。对于功能研究，我们将在急性-慢性急性-慢性模型中测试湿通路，该模型是与该联盟的小鼠模型核心共同开发的，并部分由其执行。我们将通过测量AH患者的HMGB1水平，以及分别在新的条件性HMGB1基因敲除小鼠和RAGE缺陷小鼠的两种急性-慢性AH模型中确定HMGB1及其受体RAGE的贡献来确定HMGB1在AH中的贡献(目标1)。为了确定线粒体DAMPS甲酰肽、线粒体DNA和三磷酸腺苷在急性脑出血中的作用，我们将检测患者的线粒体DNA、甲酰肽受体1(FPR1)、P2X7受体和Toll样受体9，并利用P2X7、FPR1和TLR9基因敲除小鼠(目标2)来确定它们在急性脑缺血模型中的功能贡献。对于AIM 1和AIM 2，在AH患者中测量的DAMP和DAMP受体水平也将与临床参数和该联盟项目1中产生的下一代RNA测序数据相关联。最后，我们将确定药物湿润和DAMP受体抑制对预防和治疗小鼠AH的作用，重点是抑制那些在AIMS 1和AIMS 2(AIM 3)中表现出最显著贡献的通路。我们期望揭示湿气在急性肝炎肝脏和全身炎症中的作用，并建立精选的湿气及其受体作为急性肝炎的“可用药”靶点。好了！

DESCRIPTION (provided by applicant): Alcoholic hepatitis (AH) is particularly severe form of alcoholic liver disease (ALD) that affects up to 20% of alcoholics and has a high mortality rate. However, the molecular pathways that trigger AH remain unknown, and therapy for alcoholic hepatitis has not changed over the past 30 years. Notably, targeting inflammatory cytokines such as TNF! or decreasing ROS production have been unsuccessful, and non-specific anti- inflammatory therapy with corticosteroids is only moderately effectively. In this proposal, we will investigate the hypothesis that signature molecules from dying cells, termed damage-associated molecular patterns (DAMPs), contribute to hepatic and systemic inflammation in AH, progenitor cell proliferation, and worsen liver injury and outcome. The proposed research will investigate several DAMPs with known role in sterile inflammation and neutrophil recruitment in the liver with a particular focus on (i) mitochondrial (mt) DAMPs due to their abundance in hepatoyctes and mitochondrial changes in ALD, and (ii) HMGB1, a key DAMP in necrotic liver injury. Our application will employ a combination of human studies in patients with AH and functional mouse studies, to identify key and druggable DAMP pathways with high relevance to AH. The proposed human studies will be performed in collaboration with Project 1, the Human Biorepository Core and the 10 clinical centers of the InTeam Consortium. For functional studies, we will test DAMP pathways in acute-on-chronic models of AH developed together with and performed in part by the Mouse Models Core of this consortium. We will determine the contribution of HMGB1 to AH by measuring HMGB1 levels in AH patients, and determining the contribution of HMGB1 and its receptor RAGE in two acute-on-chronic models of AH in novel conditional HMGB1 knockout mice and RAGE-deficient mice, respectively (Aim 1). To determine the contribution of mtDAMPs formyl peptide, mtDNA and ATP to AH, we will measure mtDNA, formyl peptide receptor 1 (FPR1), P2X7 receptor and Toll-like receptor 9 in patients, and determine their functional contribution in murine AH models using P2X7, FPR1 and TLR9 knockout mice (Aim 2). For both Aim 1 and Aim 2, DAMP and DAMP receptor levels measured in AH patients will be also correlated with clinical parameters and next generation RNA sequencing data generated in Project 1 of this consortium. Finally, we will determine the effect of pharmacologic DAMP and DAMP receptor inhibition for the prevention and treatment of murine AH focusing on inhibition of those pathways that showed the most significant contribution in Aims 1 and 2 (Aim 3). We anticipate to unravel the contribution of DAMPs to hepatic and systemic inflammation in AH, and to establish select DAMPs and their receptors as "druggable" targets in AH. !