Role of macrophage evolution in hepatic adaptation to alcohol.

巨噬细胞进化在肝脏适应酒精中的作用。

• 批准号: 10292924
• 负责人: STEVEN A WEINMAN
• 金额: --
• 依托单位: KANSAS CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-10-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292924
• 关键词: Acute Alcoholic Hepatitis; Affect; Alcoholic Hepatitis; Alcoholic Liver Cirrhosis; Alcoholic Liver Diseases; Alcoholic beverage heavy drinker; Alcoholic steatohepatitis; Alcohols; Animals; Anti-Inflammatory Agents; Apoptotic; Appearance; Autopsy; Awareness; Bacterial Translocation; Cell Count; Cell Death; Cells; Chronic; Cirrhosis; Complex; Data; Development; Diet; Disease; Equilibrium; Ethanol; Etiology; Evolution; Excision; Failure; Fibrosis; Gene Expression; Gene Expression Profile; Goals; Hepatic; Hepatocyte; Human; IL4 gene; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Infiltrate; Inflammatory Response; Injury; Interleukin-13; Intestines; Knowledge; Kupffer Cells; Lead; Liver; Liver diseases; Maintenance; Measures; Minority; Mus; Natural regeneration; Nature; Patients; Pattern; Phenotype; Play; Population; Prevention; Process; Production; Property; Recovery; Research; Research Personnel; Role; Severities; Signal Transduction; Source; Specimen; Techniques; Time; Tissues; Transcriptional Activation; Work; acute liver disease; alcohol exposure; base; binge drinking; cytokine; experimental study; feeding; genetic signature; hepatocyte injury; in vivo; intrahepatic; liver inflammation; liver injury; liver transplantation; macrophage; monocyte; mortality; mouse model; novel therapeutic intervention; peripheral blood; receptor; repaired; response; single-cell RNA sequencing

PROJECT TITLE: Role of macrophage evolution in hepatic adaptation to alcohol PROJECT SUMMARY / ABSTRACT Acute alcoholic hepatitis (AH) is a severe inflammatory liver disease triggered by binge drinking. The disease has a 30-day mortality of approximately 20%. One of the most striking features of AH is that it affects only a small minority of heavy drinkers suggesting that most individuals are protected from developing alcoholic liver disease by as yet unknown mechanisms. We have recently shown that alcohol exposure in mice causes early changes in liver macrophage (MΦ) populations with a loss of up to 50% of Kupffer cells and entry of infiltrating macrophages. By 10 days, Kupffer cell numbers are restored but their gene expression patterns have become more like anti-inflammatory M2 macrophages. By 35 days of alcohol exposure, the hepatic MΦ gene expression pattern changes further with a decrease in some of the classic M2 markers and the development of a gene expression pattern associated with a restorative MΦ phenotype. The macrophage population changes correlate with changes in the sensitivity of the liver to a challenge with LPS. We hypothesize that the “lost” Kupffer cells are rapidly replaced by “adaptive” macrophages leading to a state in which liver inflammation is minimal. This adaptive macrophage formation requires Kupffer cell-derived apoptotic bodies and Th2 cytokines such as IL4 and IL13. The balance of pro-and anti-inflammatory MΦs and the nature of the hepatic adaptive MΦ populations change over time so that with prolonged alcohol exposure, adaptation can be lost and liver inflammation can occur. Better understanding of the nature of the adaptive macrophages and the factors that lead to their formation, maintenance and loss would provide new approaches for the therapy of alcoholic liver disease. We will explore this hypothesis with the following specific aims: Aim 1: To determine the origins, gene expression patterns and functional properties of the mouse liver MΦ subtypes that appear after alcohol exposure. We will use lineage tracing techniques and single cell RNA sequencing to define the origin and diversity of the macrophage populations present. We will then isolate the adaptive macrophage populations to determine their functional properties both in vivo and in vitro. Aim 2: To define the signals responsible for production and maintenance of alcohol adaptive MΦ populations. We will examine the role of specific apoptotic body receptors, the impact of different sources of apoptotic bodies, timing of apoptotic body formation within the liver, and the role of hepatocyte derived factors in the formation of adaptive macrophage populations. Aim 3: To identify macrophage/monocyte populations that contribute to alcohol adaptation in humans. These experiments will leverage the mouse findings made in the first two aims to identify alcohol adaptive macrophage populations in humans. This will be done by immunohistochemical analysis of macrophages in liver autopsy specimens from non-drinkers and chronic alcohol drinkers without liver disease, and liver explants from patients with alcoholic hepatitis. These will be compared to circulating blood monocytes in these patient groups and finally will be compared with macrophages isolated from liver transplant explants of patients with alcoholic cirrhosis and cirrhosis due to etiologies unrelated to alcohol. These studies will enhance our knowledge of how macrophage phenotype changes protect the liver from alcohol, will identify the signals responsible for these changes and will define which changes protect humans from alcoholic liver disease. The long-range goal of this research is to develop macrophage directed therapies to modulate the course of alcoholic hepatitis and enhance anti- inflammatory and tissue restorative effects in a wide range of inflammatory liver diseases.

项目名称：巨噬细胞进化在肝脏适应酒精中的作用 项目概要/摘要 急性酒精性肝炎（AH）是一种由酗酒引发的严重炎症性肝病。 AH 的 30 天死亡率约为 20%，其最显着的特征之一是它仅影响患者。 少数酗酒者表明大多数人都不会患上酒精肝 我们最近发现，小鼠的酒精暴露会导致早期疾病。 肝巨噬细胞 (MΦ) 群体发生变化，库普弗细胞损失高达 50% 并进入浸润细胞 10 天后，库普弗细胞数量恢复，但其基因表达模式发生变化。 更像是抗炎M2巨噬细胞 通过酒精暴露35天，肝脏MΦ基因表达。 随着一些经典 M2 标记的减少和基因的发展，模式进一步发生变化 与恢复性 MΦ 表型相关的表达模式巨噬细胞群体变化相关。 随着肝脏对 LPS 的敏感性发生变化，我们追踪“丢失”的库普弗细胞。 迅速被“适应性”巨噬细胞取代，导致肝脏炎症最小化。 适应性巨噬细胞的形成需要库普弗细胞来源的凋亡小体和 Th2 细胞因子，例如 IL4 促炎和抗炎 MΦ 的平衡以及肝适应性 MΦ 群体的性质。 随着时间的推移而发生变化，因此，长期接触酒精，可能会丧失适应能力，并可能导致肝脏炎症 更好地了解适应性巨噬细胞的性质以及导致其发生的因素。 形成、维持和丧失将为酒精性肝病的治疗提供新的方法。 将通过以下具体目标探索这一假设： 目标 1：确定起源、基因表达 酒精暴露后出现的小鼠肝脏 MΦ 亚型的模式和功能特性。 我们将使用谱系追踪技术和单细胞 RNA 测序来定义物种的起源和多样性。 然后我们将分离适应性巨噬细胞群以确定它们的存在。 体内和体外的功能特性 目标 2：定义负责产生和释放的信号。 我们将研究特定凋亡小体的作用。 受体，不同来源的凋亡小体的影响，肝脏内凋亡小体形成的时间， 以及肝细胞衍生因子在适应性巨噬细胞群形成中的作用 目标 3： 确定有助于人类酒精适应的巨噬细胞/单核细胞群。 实验将利用前两个目标中的小鼠发现来识别酒精适应性巨噬细胞 这将通过肝脏尸检中巨噬细胞的免疫组织化学分析来完成。 来自不饮酒者和长期饮酒者但无肝病的标本，以及来自患者的肝外植体 这些将与这些患者组中的循环血液单核细胞进行比较，最后。 将与从酒精性肝硬化患者的肝移植外植体中分离出的巨噬细胞进行比较 这些研究将增强我们对巨噬细胞如何导致肝硬化的认识。 表型变化保护肝脏免受酒精侵害，将识别导致这些变化的信号，并将 确定哪些变化可以保护人类免受酒精性肝病的影响 这项研究的长期目标是 开发巨噬细胞定向疗法来调节酒精性肝炎的病程并增强抗- 在多种炎症性肝病中具有炎症和组织恢复作用。