Mitochondrial Dysfunction in the Endothelium as a Mediator of Inflammatory Injury

内皮细胞线粒体功能障碍是炎症损伤的介质

基本信息

批准号：
10706520
负责人：
Jalees Rehman
金额：
$ 42.85万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-20 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10706520
关键词：
Acute Address Affect Bacterial Proteins Binding Biogenesis Biosensor Blood Vessels Brain Cardiovascular Diseases Cell Death Cell Nucleus Cells Cellular Metabolic Process Chronic Disease Coupled Data Disease Outcome Electron Transport Endothelial Cells Endothelium Enzymes Epigenetic Process Equilibrium Functional disorder Gene Expression Profile Generations Genes Genetic Genetic Induction Genetic Models Heart Homeostasis Host Defense Human Image Immune In Vitro Individual Inflammation Inflammation Mediators Inflammatory Inflammatory Response Injury Innate Immune Response Innate Immune System Interferon Type I Knock-out Leukocytes Lung Mammalian Cell Mediator Metabolism Methionine Mitochondria Mitochondrial DNA Mitochondrial Proteins Modeling Molecular Mus Natural regeneration Nuclear Nuclear Proteins PINK1 gene Paracrine Communication Parkin Pattern Peptides Phagocytosis Phase Phenotype Phosphoric Monoester Hydrolases Play Pneumonia Production Protein Kinase Proteins Pseudomonas Reactive Oxygen Species Regulation Regulator Genes Role Signal Pathway Signal Transduction Site T-Lymphocyte TNF gene Testing Translations Ubiquitination beta catenin cell regeneration chemokine cytokine endothelial regeneration fMet-Leu-Phe receptor formyl peptide in vivo lung injury migration mitochondrial autophagy mitochondrial dysfunction neutrophil recruit response restoration transcription factor transcriptomics translatome two-photon ubiquitin ligase vascular injury

项目摘要

ABSTRACT Studies have shown that the endothelium plays a critical role in host defense by regulating the influx and phenotype of immune cells. Project 3 focuses on delineating the role of mitochondrial dysfunction and mitophagy in endothelial cells in regulating lung vascular homeostasis and host-defense during injury. The proposed studies will identify mechanisms of mitochondrial injury in ECs as well as how adaptive mitophagy activates compensatory mitochondrial biogenesis. Project 3 will also examine how mitophagy and the release of mitochondrial peptides impact neutrophils during inflammatory lung injury. The fundamental questions that will be addressed by this proposal include: What are the mechanisms of TNFα induced mitochondrial injury and mitophagy? Can we use biosensors for mitochondria in vitro and in vivo to define key phases of mitophagy and compensatory mitochondrial biogenesis? How does endothelial mitophagy affect restoration of endothelial metabolism and endothelial regeneration? How does endothelial mitophagy impact neutrophil signaling, inflammatory injury and host defense. Project 3 will test the overall hypothesis that lung endothelial mitophagy is a central regulator of endothelial homeostasis and the innate immune response. This will be addressed in two Aims. Aim 1: Determine the role of endothelial mitophagy and compensatory mitochondrial biogenesis to restore endothelial regeneration and homeostasis during inflammatory lung injury. Here we will test the hypothesis that inflammation induces mitochondrial injury and adaptive mitophagy in the lung endothelium to orchestrate endothelial regeneration and homeostasis. Furthermore, we posit that the central mechanism of restoring homeostasis is through the compensatory mitochondrial biogenesis. We use intravital two photon imaging of endothelial mitophagy (with Core D) and perform in vivo studies using EC- specific genetic deletion of the mitophagy mediator PINK1, the mitochondrial biogenesis transcription factors PGC1α and TFAM, and the mitochondrial phosphatase PGAM5 (which transfers post-mitophagy signaling to the nucleus) with mechanistically driven in vitro studies in human lung ECs. These studies will be coupled to analysis of EC metabolism, mitochondrial biogenesis, and EC regeneration. Aim 2: Determine the role of endothelial mitophagy in activating the lung host-defense function during inflammatory injury. Here we will test the hypothesis that EC-mitophagy increases transendothelial neutrophil influx and thereby bacterial killing through secretion of formylated peptides and activation of formyl peptide receptors. We will use genetic models of EC-specific PINK1 deletion in distinct models of inflammatory lung injury (LPS and Pseudomonas pneumonia) and analyze EC epigenetic and transcriptomic regulation (with Core B) wrought by mitophagy and address how these contribute to host-defense function of EC and neutrophils downstream of EC PINK1. We will also assess how EC-PINK1 regulates neutrophil-induced lung injury and generation of N-formylated mitochondrial peptides. We will also determine the effects of lung EC mitophagy on neutrophil transmigration and bacterial phagocytosis (with Core D) and study the signaling pathways (with Core C) downstream of mitochondrial formyl peptides in neutrophils.

抽象的研究表明，内皮细胞通过调节流入和表型在宿主防御中发挥关键作用项目 3 重点描述线粒体功能障碍和线粒体自噬在内皮细胞中的作用。拟议的研究将确定细胞在损伤期间调节肺血管稳态和宿主防御的作用。 EC 中线粒体损伤的机制以及适应性线粒体自噬如何激活补偿性线粒体项目 3 还将研究线粒体自噬和线粒体肽的释放如何影响中性粒细胞。该提案将解决的基本问题包括： TNFα 诱导线粒体损伤和线粒体自噬的机制是什么？我们可以使用线粒体生物传感器吗？体外和体内确定线粒体自噬和补偿性线粒体生物发生的关键阶段是如何进行的？内皮线粒体自噬如何影响内皮代谢的恢复和内皮再生？内皮线粒体自噬对中性粒细胞信号传导、炎症损伤和宿主防御的影响项目 3 将进行测试。总体假设是肺内皮线粒体自噬是内皮稳态和先天性的中央调节器这将在两个目标中得到解决：确定内皮线粒体自噬和免疫反应的作用。补偿性线粒体生物发生以恢复内皮再生和稳态在这里，我们将检验炎症诱导线粒体损伤的假设。肺内皮细胞的适应性线粒体自噬协调内皮再生和稳态。我们认为恢复体内平衡的核心机制是通过补偿性线粒体生物发生。我们使用内皮线粒体自噬的活体双光子成像（使用 Core D）并使用 EC- 进行体内研究线粒体自噬介体 PINK1（线粒体生物合成转录因子 PGC1α）的特异性基因缺失和 TFAM，以及线粒体磷酸酶 PGAM5（将线粒体自噬后信号传递至细胞核）对人肺 EC 进行机械驱动的体外研究，这些研究将与 EC 的分析相结合。代谢、线粒体生物发生和 EC 再生目标 2：确定内皮线粒体自噬的作用。在炎症损伤期间激活肺宿主防御功能在这里我们将检验以下假设： EC-线粒体自噬增加跨内皮中性粒细胞流入，从而通过分泌我们将使用 EC 特异性 PINK1 的遗传模型。不同炎症性肺损伤模型（LPS 和假单胞菌肺炎）中的缺失并分析 EC 线粒体自噬引起的表观遗传和转录组调控（核心 B）并解决了这些调控如何促进线粒体自噬 EC 和 EC PINK1 下游中性粒细胞的宿主防御功能我们还将评估 EC-PINK1 如何调节。我们还将确定中性粒细胞诱导的肺损伤和 N-甲酰化线粒体肽的生成。肺 EC 线粒体自噬对中性粒细胞迁移和细菌吞噬作用的影响（核心 D）并研究中性粒细胞中线粒体甲酰基肽下游的信号通路（具有核心 C）。