Mechanism of neutrophilic NCF1 in alcohol-associated liver disease pathogenesis

中性粒细胞NCF1在酒精相关性肝病发病机制中的作用机制

• 批准号: 10723321
• 负责人: JING MA
• 金额: $ 14.72万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-08 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10723321
• 关键词: Address; Alcoholic Hepatitis; Alcoholic Liver Diseases; Animal Model; Automobile Driving; Basic Science; Bilirubin; Binding; Biogenesis; Cell Communication; Cell model; Cells; Cellular Stress; Clinical; Complex; Data; Disease; Docking; Ethanol; Generations; Genes; Hepatic; Hepatitis; Hepatocyte; Homeostasis; Immune; Inflammasome; Inflammation Mediators; Inflammatory Response; Knock-out; Knowledge; Link; Liver; Liver diseases; MAP Kinase Gene; Mediating; Medical; Membrane; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Multienzyme Complexes; Mus; Myeloid Cells; NADP; Neutrophil Infiltration; Oxidases; Oxidative Stress; Oxidative Stress Induction; Pathogenesis; Pathway interactions; Patients; Pattern; Phagocytes; Phenotype; Phosphotransferases; Play; Production; Proteins; Public Health; Reactive Oxygen Species; Research Activity; Role; Serum; Severity of illness; Signal Pathway; Translational Research; United States; Up-Regulation; alcohol use disorder; cell injury; end stage liver disease; experimental study; extracellular; extracellular vesicles; hepatocyte injury; improved; intercellular communication; interpatient variability; intrahepatic; liver injury; mortality; nanoscale; neutrophil; p38 Mitogen Activated Protein Kinase; systemic inflammatory response; targeted treatment; tissue injury; transcriptome sequencing; vesicular release

Project Summary Alcohol-associated liver disease (ALD) is a complex disease and represents a spectrum of histopathological changes in the liver. Accumulating evidence suggests that multiple types of immune cells are involved in the pathogenesis of alcohol-associated hepatitis (AH), particularly neutrophils. However, the mechanisms underlying neutrophils in mediating AH pathogenesis are not well understood. Our exploratory experiments had led to an important and clinical observation on the inter-patient variability and two distinct patterns of hepatic neutrophil infiltration. Our data also indicated that higher level of hepatic parenchymal neutrophils was associated with AH disease severity. The key question we would like to address in this application is how neutrophils drives disease severity and mediates liver injury in ALD. Mechanistically, neutrophils, as an innate inflammatory response, mediates tissue injury by producing inflammatory mediators and reactive oxygen species (ROS). ROS production in neutrophils is regulated by the multi-meric transmembrane enzyme complex, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX). Among them, the NCF1 gene, which encodes the phagocytic oxidase (phox) unit p47phox, is predominantly expressed in neutrophils and plays an important role in controlling ROS production in neutrophils. Our data suggested that miR-223, the most abundant neutrophilic miRNA, may be a downstream target of NCF1 and its induction ameliorates alcohol-induced liver injury. In specific aim#1, we will determine the molecular mechanism on the inhibition of neutrophilic miR-223 by NCF1. Our data and planned experiments in specific aim#1 will expand our current knowledge on the signaling pathway in the neutrophils, neutrophilic Ncf1-induced oxidative stress, and miR- 223 expression. The next important question to address is how the inhibition of miR-223 by NCF1-induced oxidative stress leads to hepatocyte injury. In specific aim #2, we will determine the downstream crosstalk between neutrophil-induced ROS generation and hepatocytes via extracellular vesicles (EVs) leading to alcohol-induced liver injury. Taken together, we have developed animal and cellular models to mechanistically examine the roles of neutrophils in ALD pathogenesis. Understanding the mechanism linking neutrophils to ALD pathogenesis is of importance; this will pave a way for the discovery of potential targeted therapy for patients with ALD.

项目概要 酒精相关性肝病（ALD）是一种复杂的疾病，代表了一系列组织病理学特征 肝脏的变化。越来越多的证据表明多种类型的免疫细胞参与了 酒精相关性肝炎（AH）的发病机制，特别是中性粒细胞。然而，这些机制 中性粒细胞在介导 AH 发病机制中的作用尚不清楚。我们的探索性实验 导致了对患者间变异性和两种不同模式的重要临床观察 肝脏中性粒细胞浸润。我们的数据还表明，肝实质中性粒细胞水平较高 与 AH 疾病的严重程度相关。我们想在这个应用程序中解决的关键问题是如何 中性粒细胞会导致 ALD 疾病严重程度并介导肝损伤。从机制上讲，中性粒细胞作为一种先天的 炎症反应，通过产生炎症介质和活性氧介导组织损伤 物种（ROS）。中性粒细胞中 ROS 的产生受多聚跨膜酶的调节 复合物，烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶（NOX）。其中，NCF1 编码吞噬氧化酶 (phox) 单位 p47phox 的基因主要在中性粒细胞中表达 并在控制中性粒细胞中ROS的产生中发挥重要作用。我们的数据表明 miR-223， 最丰富的中性粒细胞 miRNA，可能是 NCF1 的下游靶点及其诱导改善 酒精引起的肝损伤。在具体目标#1中，我们将确定抑制的分子机制 NCF1 影响中性粒细胞 miR-223。我们在特定目标#1 中的数据和计划实验将扩展我们当前的 中性粒细胞信号通路、中性粒细胞 Ncf1 诱导的氧化应激和 miR- 223 表达。下一个要解决的重要问题是NCF1如何诱导miR-223的抑制 氧化应激导致肝细胞损伤。在具体目标#2中，我们将确定下游串扰 中性粒细胞诱导的 ROS 生成和肝细胞之间通过细胞外囊泡 (EV) 产生 酒精引起的肝损伤。总而言之，我们开发了动物和细胞模型，以机械方式 检查中性粒细胞在 ALD 发病机制中的作用。了解中性粒细胞与 ALD 发病机制很重要；这将为发现潜在的靶向治疗铺平道路 酒精性肝病（ALD）患者。