Neutrophil Priming in Trauma and Sepsis

创伤和脓毒症中的中性粒细胞启动

• 批准号: 7933278
• 负责人: MICHAEL B YAFFE
• 金额: $ 24.13万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7933278
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute; Acute Lung Injury; Adult; Adult Respiratory Distress Syndrome; Binding; Biochemical; Biochemistry; Cause of Death; Cells; Cellular biology; Complex; Critical Illness; Cytochromes; Cytoplasm; Cytoskeleton; Cytosol; Data; Development; Event; Funding; Goals; Human; Inflammatory; Injury; Intensive Care Units; Knowledge; Lipids; MAP Kinase Gene; MAPK14 gene; Mediating; Membrane; Modeling; Molecular; Monomeric GTP-Binding Proteins; Morbidity - disease rate; Mus; NADPH Oxidase; Organ failure; Oxidases; Pathogenesis; Pathway interactions; Patients; Phagocytes; Phosphotransferases; Play; Prevention strategy; Process; Production; Protein Binding Domain; Protein Kinase; Proteins; Public Health; Reactive Oxygen Species; Reagent; Regulation; Research; Respiratory Burst; Rest; Risk; Role; Sepsis; Series; Signal Pathway; Superoxides; Surgical Intensive Care; Syndrome; Therapeutic; Therapeutic Agents; Tissues; Trauma; Work; base; cytochrome b558; cytokine; design; extracellular; human CYBA protein; human disease; improved; in vivo; killings; lung injury; microbial; monomer; mortality; mouse model; neutrophil; neutrophil cytosol factor 40K; neutrophil cytosol factor 67K; novel diagnostics; pathogen; protein protein interaction

DESCRIPTION (provided by applicant): Neutrophil priming and activation following trauma and sepsis is a key event implicated in causing Adult Respiratory Distress Syndrome (ARDS), Acute Lung Injury (ALI) and Multi-Organ Failure Syndrome (MOSF). Priming of the respiratory burst by cytokines following injury and sepsis results in excessive superoxide production by the NADPH oxidase leading to auto-inflammatory tissue damage. Many of the molecular mechanisms involved in priming and activation of the NADPH oxidase, however, remain poorly defined. Our long-term goal is to develop a detailed molecular understanding of how protein kinase and lipid kinase signaling pathways, including the PI 3-kinase pathway, the p38MAPK pathway and the Erk1/2 pathways regulate the assembly, subcellular targeting, and activity of the neutrophil NADPH oxidase during priming and activation. Our previous work and preliminary observations identified PX domains in the p47phox and p40phox subunits as modular protein domains that bind to specific lipid products of PI 3-kinase, and showed that the priming agents PAF and TNF1 induced the assembly of a p47phox:p67phox:p40phox heterotrimeric complex in the cytoplasm of primed but un-activated cells. In the studies outlined in this proposal we investigate the importance of PX domain-mediated interactions with lipids and the cytoskeleton, and p47phox binding to PDZ- domain-containing proteins, in neutrophil priming and activation using biochemistry, cell biology and mouse models. In addition, we further explore protein kinase-dependent molecular mechanisms involved in heterotrimer formation during priming. The results from these studies may assist in the development of novel diagnostic or therapeutic reagents aimed at limiting the auto-inflammatory tissue damage patients suffer as a result of sepsis and trauma. PUBLIC HEALTH RELEVANCE3 Neutrophil priming and activation following trauma and sepsis is a key event implicated in causing Adult Respiratory Distress Syndrome (ARDS), Acute Lung Injury (ALI) and Multi-Organ Failure Syndrome (MOSF), which are the leading causes of death in adult surgical intensive care units. Priming of the neutrophil respiratory burst by cytokines following injury and sepsis results in excessive superoxide production by the NADPH oxidase and contributes to the auto-inflammatory tissue damage seen in these syndromes. Our research is designed to better understand regulation of the NADPH oxidase following trauma and sepsis, and facilitate the development of novel diagnostic and therapeutic agents that will reduce tissue damage, morbidity and mortality in these critically ill patients.

描述（由申请人提供）：创伤和败血症后的中性粒细胞启动和激活是一个关键事件，涉及引起成人呼吸窘迫综合征（ARDS），急性肺损伤（ALI）和多器官衰竭综合征（MOSF）。损伤和败血症后细胞因子爆发的呼吸爆发会导致NADPH氧化酶产生过度的超氧化物，从而导致自动炎性组织损伤。然而，许多参与NADPH氧化酶启动和激活的分子机制仍然很差。我们的长期目标是对蛋白激酶和脂质激酶信号传导途径（包括Pi 3-激酶途径，p38MAPK途径以及ERK1/2途径）的详细分子了解如何调节组装，亚细胞靶向和中性粒细胞NADPH氧化酶的活性。 Our previous work and preliminary observations identified PX domains in the p47phox and p40phox subunits as modular protein domains that bind to specific lipid products of PI 3-kinase, and showed that the priming agents PAF and TNF1 induced the assembly of a p47phox:p67phox:p40phox heterotrimeric complex in the cytoplasm of primed but un-activated细胞。在该提案中概述的研究中，我们研究了PX结构域介导的与脂质与细胞骨架的相互作用的重要性，以及使用生物化学，细胞生物学和小鼠模型在中性粒细胞启动和激活中与含PDZ-域的蛋白质的P47Phox结合。此外，我们进一步探讨了启动过程中参与异构三聚体形成的蛋白激酶依赖性分子机制。这些研究的结果可能有助于开发新的诊断或治疗试剂，旨在限制自身炎性组织损害患者因脓毒症和创伤而遭受的损失。公共卫生相关3嗜中性粒细胞启动和创伤和败血症后的激活是一个关键事件，涉及引起成人呼吸窘迫综合征（ARDS），急性肺损伤（ALI）和多器官衰竭综合征（MOSF），它们是成人外科手术强化护理部门的主要死亡原因。损伤和败血症后细胞因子爆发中性粒细胞呼吸道爆发会导致NADPH氧化酶产生过多的超氧化物，并导致这些综合征中看到的自动炎性组织损伤。我们的研究旨在更好地了解创伤和败血症后NADPH氧化酶的调节，并促进新型诊断和治疗剂的发展，从而降低这些重症患者的组织损伤，发病率和死亡率。