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: 10515320
• 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/10515320
• 关键词: Acute Alcoholic Hepatitis; Affect; Alcoholic Hepatitis; Alcoholic Liver Cirrhosis; Alcoholic Liver Diseases; Alcoholic beverage heavy drinker; Alcoholic steatohepatitis; Alcohols; Animal Experiments; Animals; Anti-Inflammatory Agents; Apoptotic; Appearance; Autopsy; Awareness; Bacterial Translocation; Cell Count; Cell Death; Cells; Cessation of life; Chronic; Cirrhosis; Complex; Data; Development; Diet; Disease; Equilibrium; Etiology; Evolution; Excision; Failure; Fibrosis; Gene Expression Profile; Goals; Hepatic; Hepatocyte; Human; IL4 gene; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Inflammatory Infiltrate; Inflammatory Response; Injury; Interleukin-13; Intestines; Knowledge; Kupffer Cells; Liver; Liver diseases; Macrophage; Maintenance; Measures; Minority; Mus; Natural regeneration; Nature; Patients; Pattern; Phenotype; Play; Population; Prevention; Process; Production; Property; Recovery; Research; Research Personnel; Resolution; Role; Severities; Signal Transduction; Source; Specimen; Techniques; Time; Tissues; Transcriptional Activation; Work; acute liver disease; alcohol exposure; binge drinking; cytokine; experimental study; feeding; genetic signature; hepatocyte injury; in vivo; intrahepatic; liver inflammation; liver injury; liver transplantation; monocyte; mortality; mouse model; novel therapeutic intervention; peripheral blood; receptor; repaired; response; single-cell RNA sequencing; tissue injury

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：巨噬细胞演变在肝适应酒精中的作用 项目摘要 /摘要 急性酒精性肝炎（AH）是一种严重的炎症性肝病，是由暴饮暴食触发的。疾病 30天死亡率约为20％。 AH最引人注目的特征之一是它仅影响一个 少数少数饮酒者建议大多数人都受到保护，免受饮酒肝的发展 疾病尚未清楚的机制。我们最近表明，小鼠的酒精暴露会早期引起 肝巨噬细胞（Mφ）种群的变化，损失高达50％的库普弗细胞和渗透的进入 巨噬细胞。到10天，库普夫细胞数已经恢复，但它们的基因表达模式已成为 更像是抗炎的M2巨噬细胞。到35天的酒精暴露，肝Mφ基因表达 图案随着某些经典M2标记和基因的发展而进一步变化 与恢复性Mφ表型相关的表达模式。巨噬细胞的变化相关 随着肝脏对LPS挑战的敏感性的变化。我们假设“丢失”的库普弗细胞 迅速被“自适应”巨噬细胞替代，导致肝脏注射最少的状态。 自适应巨噬细胞形成需要库普弗细胞衍生的凋亡人体和Th2细胞因子（例如IL4） 和IL13。抗炎和抗炎的平衡以及肝自适应Mφ种群的性质 随着时间的流逝，随着时间的流逝，随着酒精的长时间暴露，适应可能会丢失，肝脏炎症可以 发生。更好地理解自适应巨噬细胞的性质以及导致其自适应的因素 形成，维持和损失将为酒精性肝病治疗提供新的方法。我们 将以以下特定目的探索这一假设：目标1：确定起源，基因表达 酒精暴露后出现的小鼠肝Mφ亚型的模式和功能特性。 我们将使用谱系跟踪技术和单细胞RNA测序来定义 存在巨噬细胞种群。然后，我们将分离自适应巨噬细胞以确定其 体内和体外的功能性能。目标2：定义负责生产的信号和 维持酒精自适应Mφ种群。我们将检查特定凋亡人体的作用 受体，凋亡人物不同来源的影响，肝脏内凋亡人体形成的时机， 肝细胞衍生因子在形成自适应巨噬细胞种群中的作用。目标3：到 确定巨噬细胞/单核细胞种群，导致人类的酒精适应。这些 实验将利用前两个目的中的小鼠发现来识别酒精自适应巨噬细胞 人类的人口。这将通过对肝尸检中巨噬细胞的免疫组织化学分析来完成 来自无肝病的非饮酒者和慢性酒精饮用者的标本，以及患者的肝外植体 与酒精性肝炎。这些将与这些患者组的循环血单核细胞进行比较，最后 将将酒精性肝硬化患者和 由于病因与酒精无关的肝硬化。这些研究将增强我们对巨噬细胞的了解 表型变化可保护肝脏免受酒精的影响，将确定负责这些变化的信号，并将 定义改变人类免受酒精性肝病的影响。这项研究的远程目标是 开发巨噬细胞定向疗法以调节酒精性肝炎的病程并增强抗 在多种炎症性肝病中的炎症和组织修复作用。