Alcohol Promotes Hepatitis B Progression by Impairment of Innate Immunity in Liver Cells

这个项目是关于曝光组和酒精的P50资助的一部分。它专注于理解先天的 免疫机制，通过环境暴露乙醇代谢物增加水平 乙肝病毒(乙肝)感染，并促进其持久性和肝脏损伤。尽管有预防性疫苗 据估计，全球慢性乙肝病毒携带者的数量高达2.5亿人，其中 年死亡率为80万。慢性乙肝病毒感染率、病毒载量及预后 终末期疾病在滥用酒精的乙肝患者中更高，但背后的机制 事件仍不清楚。在这里，我们将研究接触病毒、酒精和不饱和脂肪酸如何影响 肝细胞对干扰素的反应。我们还将研究巨噬细胞对小鼠心脏的保护作用。 干扰素刺激基因(ISGs)的激活和肝细胞中乙肝病毒水平的降低。一种抗病毒药物 ISGS，APOBEC3G，因其能降解HBVccDNA而具有特殊的重要性。巨噬细胞--肝细胞 细胞外小泡(EVS)是细胞外信息传递的媒介。然而，电动汽车的保护作用 当细胞暴露在酒精中时，来自巨噬细胞的物质可能会被破坏。高脂肪饮食进一步加重肝炎 酒精增强了B的致病作用。我们的中心假设是：接触乙醇代谢物会损害 乙肝病毒感染的肝细胞和病毒激活的巨噬细胞中的干扰素信号，通过限制 抗病毒ISG诱导，以及通过破坏EV介导的保护性串扰间接地 巨噬细胞和肝细胞，从而促进病毒在肝内扩散，增强致病作用。这个 乙醇对表达乙肝病毒的肝细胞天然免疫的抑制作用可能进一步增强 细胞暴露于不饱和脂肪酸。在这项研究中，我们提出了三个目标： 目的1：研究RIG1-MAVS-1激活ISGs对肝细胞内乙肝病毒感染的调节作用。 IRF3或cGAS-STING-IRF3和JAK-STAT1/2途径在乙醇代谢产物和 不饱和脂肪酸。 目的2：在酒精和不饱和脂肪酸暴露的情况下，评估EV介导的串扰 在乙肝病毒感染的肝细胞和巨噬细胞之间，通过天然免疫调节抗病毒保护。 目的3：阐明酒精和高脂饮食对乙肝病毒表达、免疫球蛋白诱导和肝脏的影响。 转基因复制乙肝病毒小鼠的脂肪变性/炎症，并研究这些暴露是否会影响其他 这些小鼠的器官/组织(肺、心、脾、胰腺、脂肪组织)。 所获得的信息有很强的潜力转化为临床实践和干预措施，通过 先天免疫功能障碍生物标志物的研究进展及可能治疗方法的确定 目标。

This project is a part of a P50 grant on exposome and alcohol. It is focused on understanding the innate immunity mechanisms, by which environmental exposure to ethanol metabolites increases the levels of hepatitis B viral (HBV) infection and promotes its persistence and liver injury. Despite a prophylactic vaccine availability, the number of chronic HBV carriers is estimated to be as high as 250 million worldwide, with an annual death rate of 800,000. The incidences of chronic HBV -infection, the viral load and the outcomes of end-stage diseases are higher in hepatitis B patients abusing alcohol, but the mechanisms behind these events are still unclear. Here, we will study how exposures to virus, alcohol and unsaturated fatty acids affect interferon (IFN) response in hepatocytes. We will also study the protective effect of macrophages on the activation of interferon-stimulated genes (ISGs) and the reduction of HBV levels in hepatocytes. One anti-viral ISGs, APOBEC3G, is of special importance since it degrades HBV cccDNA. This macrophage-hepatocyte communication is mediated via extracellular vesicles (EVs). However, the protective effects of EVs secreted from macrophages may be ruined when cells are exposed to alcohol. High-fat diet further exacerbates hepatitis B pathogenesis enhanced by alcohol. Our central hypothesis is: that exposure to ethanol metabolites impairs interferon signaling in HBV-infected hepatocytes and virus-activated macrophages both directly, via limiting anti-viral ISG induction, and indirectly, via disrupting the protective EV-mediated crosstalk between macrophages and hepatocytes, thereby promoting intrahepatic viral spread and enhanced pathogenesis. The suppressive effects of ethanol on innate immunity in HBV-expressing hepatocytes may be further enhanced by cell exposure to unsaturated fatty acids. For this study, we proposed 3 Aims: Aim 1: To study the regulation of HBV-infection in hepatocytes by the activation of ISGs via the RIG1-MAVS- IRF3 or the cGAS-STING-IRF3 and the JAK-STAT1/2 pathways under exposure to ethanol metabolites and unsaturated fatty acids. Aim 2: In the settings of alcohol and unsaturated fatty acid exposures, to assess the EV-mediated crosstalk between HBV-infected hepatocytes and macrophages, which regulates anti-viral protection by innate immunity. Aim 3: To elucidate the contribution of ethanol and high-fat diet to HBV expression, ISG induction, and liver steatosis/inflammation in transgenic HBV-replicating mice, and to study whether these exposures affect other organs/tissues (lung, heart, spleen, pancreas, adipose tissue) in these mice. The obtained information has the strong potential to be translated to clinical practice and interventions through the development of biomarkers for innate immunity dysfunction and the identification of possible treatment targets.