The Trojan Horse Hypothesis: Neutrophil Elastase Reprograms Macrophage Function

特洛伊木马假说：中性粒细胞弹性蛋白酶重新编程巨噬细胞功能

基本信息

批准号：
10683401
负责人：
Judith A Voynow
金额：
$ 46.95万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10683401
关键词：
Acetylation Biological Assay Biological Markers Cell Nucleus Cell Survival Cells Chromatin Chromatin Structure Citrulline Confocal Microscopy Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Cytoplasm Cytoplasmic Organelle Cytoplasmic Tail DNA Data Disease Progression Equus caballus Failure Gender Gene Expression Genetic Transcription Glutathione Disulfide HMGB1 gene Histone Deacetylase Histone H3 Human Infection Control Inflammation Inflammatory Inflammatory Response Interruption Leukocyte Elastase Lung diseases Lysine Macrophage Maps Mitochondria Modification Molecular Morbidity - disease rate Natural Immunity Nuclear Pathway interactions Pattern Peptide Hydrolases Phagocytosis Phenotype PicoGreen Production Proteins Reactive Oxygen Species SOD2 gene Signal Induction Signal Pathway Sirtuins Small Interfering RNA Source Stimulus Structure Superoxide Dismutase TNF gene Testing Up-Regulation airway inflammation cystic fibrosis airway cytokine extracellular healthy volunteer histone deacetylase 2 knock-down monocyte mortality new therapeutic target novel programs

项目摘要

PROJECT SUMMARY In Cystic Fibrosis (CF), airway innate immunity is breached. Macrophage phagocytosis and efferocytosis fail, and macrophages augment airway inflammation by production of excess cytokines and release of High Mobility Group Box 1 (HMGB1), a damage associated molecular pattern. Although loss of CFTR impacts macrophage function, the CF airway milieu, which is typified by micromolar concentrations of neutrophil elastase (NE), is an overwhelming stimulus, activating a robust pro-inflammatory response in macrophages from both CF and healthy subjects. We propose that NE, a biomarker for lung disease progression in CF subverts macrophage function from protective to pro- inflammatory, yet the mechanisms by which NE reprograms the macrophage are not completely understood. In this application, we present an unprecedented hypothetical mechanism to explain how NE alters macrophage function. Extracellular NE is rapidly endocytosed by the macrophage. But instead of being degraded, proteolytically active NE is localized to both cytoplasmic domains and the nucleus, resulting in increased cytokine expression and release of macrophage extracellular traps (METs). Thus, NE functions like a Trojan Horse to subvert macrophage function. We will test this hypothesis in primary human blood monocyte derived macrophages from healthy volunteers and subjects with CF. The Specific Aims follow: Aim 1. To determine whether NE degrades HDACs, resulting in acetylation of downstream targets, leading to transcriptional upregulation of TNFα and release of HMGB1. Aim 2. To evaluate whether NE protease activity and/or NE-generated reactive oxygen species increase release of mitochondrial and nuclear METs. RELEVANCE: Results from this project will define a novel mechanism utilized by NE to promote sustained airway inflammation, the major cause of morbidity and mortality in CF, and identify targets for new therapies to interrupt the relentless progression of lung disease.

项目摘要在囊性纤维化（CF）中，气道先天免疫受到破坏。巨噬细胞吞噬作用和递增细胞增多症失败，巨噬细胞通过产生多余的细胞因子增强气道注入和释放高迁移率组框1（HMGB1），损伤相关的分子模式。虽然CFTR的损失影响巨噬细胞函数，CF气道环境，该环境的特征是微摩尔浓度的中性粒细胞弹性酶（NE）是一种压倒性的刺激，激活了强大的促炎反应在CF和健康受试者的巨噬细胞中。我们建议NE，一种肺部疾病的生物标志物 CF的进展颠覆了巨噬细胞的功能，从受保护到炎症性，但是 NE重编程巨噬细胞的机制尚未完全理解。在此应用中，我们提出了一种前所未有的假设机制，以解释NE如何改变巨噬细胞功能。细胞外NE被巨噬细胞迅速内吞。但是不是降解，蛋白水解活性NE位于细胞质结构域和细胞核上，导致在增加的细胞因子表达和巨噬细胞外陷阱（MetS）的释放中。那，ne 功能像木马马一样颠覆巨噬细胞功能。我们将在初级中检验这一假设人类血液单核细胞从健康的志愿者和CF受试者中衍生出巨噬细胞。具体目的如下：目标1。确定NE是否降低HDAC，导致下游靶标的乙酰化，导致 TNFα的转录上调和HMGB1的释放。目的2。评估NE蛋白酶活性和/或NE产生的活性氧增加线粒体和核MET的释放。相关性：该项目的结果将定义NE使用的新机制来促进持续的气道注入，是CF发病率和死亡率的主要原因，并确定中断肺部疾病远程进展的新疗法。