Inhibition of Sterile Inflammation by Digoxin in Alcoholic Hepatitis

摘要：酒精性肝炎（AH）是一种主要的肝脏疾病，住院死亡率在25- 40%之间。 肝脏炎症是AH的一个关键特征，但驱动这种炎症反应的因素尚不清楚。 我们已经确定了肝脏炎症的新的关键驱动因素，这是本申请的主题。我们还 确定了新的蛋白质组学和分子标记物在AH，这将用于预测预后。 我们已经证明，激活核（缺氧抑制因子1-α：HIF-1α）途径是必需的， 对于持续性无菌性炎症的发展，这表明抑制HIF-1α可能是 治疗中的AH[1]。在高通量筛选中，强心苷被鉴定为具有显著的能力， 抑制HIF-1α[2]。初步数据表明i）肝组织中HIF-1α依赖性基因的上调 与重度AH相比，早期AH，来自InTeam联盟ii）地高辛减少组织 酒精模型中的损伤，以及其他形式的肝损伤。iii）地高辛与丙酮酸激酶结合 iv）地高辛减少PKM 2与HIF-1α启动子的结合并限制HIF-1α的上调，和 HIF-1α反应基因。v）基于适体的血清蛋白质组学分析显示，在AH患者中， 和全身炎症反应（SIRS）中存在肿瘤坏死因子相关蛋白的增加， 低亲和力免疫球蛋白γ Fc区受体II、补体成分、激肽释放酶和成纤维细胞 生长因子vi）已知血清DNA是促炎性配体，并且血清DNA水平与 AH患者外周血白色细胞计数。 目标1。获得支持地高辛治疗酒精性肝炎的临床数据。 目标二。确定酒精性肝炎中PKM 2调控的主要和新靶点。 目标3：获得血浆蛋白质组学和分子数据，以便早期识别SIRS患者。 总的来说，这将使我们能够获得必要的数据，用于在AH中临床测试低剂量地高辛。在 此外，它将使我们能够在患者血清中识别新的蛋白质标记物和促炎信号 与AH。最后，我们将能够确定是否有任何新的蛋白质标记物与新的PKM 2相关 我们发现的路径。

ABSTRACT: Alcoholic hepatitis (AH) is a major liver disease, and has an inpatient mortality of between 25-40%. Liver inflammation is a key feature of AH, yet the factors which drive this inflammatory response are not known. We have identified novel key drivers of liver inflammation which are the subject of this application. We have also identified novel proteomic and molecular markers in AH which will be used to predict prognosis. We have shown that activation of the nuclear (hypoxia inhibitory factor 1-α: HIF-1α) pathway was required for the development of sustained sterile inflammation, which suggested that inhibition of HIF-1α may be therapeutic in AH[1]. In a high throughput screen cardiac glycoside were identified to have significant ability to inhibit HIF-1α[2]. The preliminary data demonstrates i) Up-regulation of HIF-1α dependent genes in liver tissues from early AH, as compared to severe AH, from the InTeam consortium ii) Ability of digoxin to reduce tissue damage in a model of alcohol, and others forms of liver injury. iii) Digoxin binds to the enzyme pyruvate kinase M2 (PKM2), iv) Digoxin reduces PKM2 binding to the HIF-1α promoter and limits up-regulation of HIF-1α, and HIF-1α response genes. v) An aptamer based proteomic analysis of serum shows that in patients with the AH and the systemic inflammatory response (SIRS) there is an increase in tumor necrosis factor related proteins, low affinity immunoglobulin gamma Fc region receptor II, complement components, kallikrein and fibroblast growth factors. vi) Serum DNA is known to be a pro-inflammatory ligand and serum DNA levels correlated with peripheral blood white cell count in AH. Aim 1. Obtain clinical data supporting the therapeutic use of digoxin in alcoholic hepatitis. Aim 2. Identify dominant and novel targets that are regulated by PKM2 in alcoholic hepatitis. Aim 3. Obtain plasma proteomic and molecular data to allow for early identification of patients with SIRS. Collectively this will allow us to obtain the necessary data towards clinically testing low dose digoxin in AH. In addition, it will allow us to identify novel protein markers and pro-inflammatory signals in the serum of patients with AH. Finally, we will be able to identify if any of the novel protein markers are associated with the novel PKM2 pathway we have identified.