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

• 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
• 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

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，内皮通过调节影响和表型在宿主防御中起关键作用 免疫细胞。项目3的重点是描述内皮中线粒体功能障碍和线粒体的作用 调节肺血管内稳态和宿主防御的细胞受伤。拟议的研究将确定 EC中线粒体损伤的机制以及自适应线粒体如何激活补偿性线粒体 生物发生。项目3还将研究线粒体胡椒体的释放和释放中性粒细胞 在炎症性肺部受伤期间。该提案将解决的基本问题包括：什么 TNFα的机制是否诱导线粒体损伤和线粒体？我们可以将生物传感器用于线粒体吗 体外和体内定义线粒体和补偿性线粒体生物发生的关键阶段？怎么样 内皮线粒体会影响内皮代谢和内皮再生的恢复？怎么样 内皮线粒体影响中性粒细胞信号传导，炎症性损伤和宿主防御。项目3将测试 总体假设肺内皮线粒体是内皮稳态和先天性的中心调节者 免疫反应。这将在两个目标中解决。目标1：确定内皮线粒体的作用和 补偿性线粒体生物发生以恢复内皮再生和稳态的生物发生 炎症性肺损伤。在这里，我们将检验以下假设：炎症会诱导线粒体损伤和 在肺部原始植物中自适应线索，以策划原始的再生和稳态。此外， 我们认为恢复体内稳态的核心机制是通过补偿性线粒体生物发生。 我们使用内皮线皮的两种光子成像（带有核心D），并使用EC-进行体内研究 线粒体生物发生转录因子PGC1α的特定遗传缺失 和TFAM，以及线粒体磷酸酶PGAM5（将线粒体后信号传递到细胞核） 通过机械驱动的人类肺EC的体外研究。这些研究将与EC的分析结合 代谢，线粒体生物发生和EC再生。目标2：确定内皮线粒体的作用 在炎症损伤期间激活肺部防御功能。在这里，我们将检验以下假设 ec-mitophagy增加了跨内皮性嗜中性粒细胞的影响，从而通过分泌细菌杀死细菌 甲基化的宠物和宠物受体的激活。我们将使用EC特异性PINK1的遗传模型 在炎症性肺损伤（LPS和PSEUDOMONAS肺炎）的不同模型中缺失并分析EC 表观遗传学和转录组调节（核心B）通过线粒体包裹，并解决这些贡献 EC Pink1下游EC和中性粒细胞的宿主防御功能。我们还将评估EC-Pink1如何调节 嗜中性粒细胞诱导的肺损伤和N型线粒体胡椒体的产生。我们还将确定 肺部线粒体对中性粒细胞传播和细菌吞噬作用（核心D）的影响并研究 信号通路（带有核心C）在中性粒细胞中的线粒体金酰基下游。