Pulmonary Collectins, Hyaluronan and Macrophages

肺集合素、透明质酸和巨噬细胞

基本信息

批准号：
7279188
负责人：
RASHMIN C SAVANI
金额：
$ 32万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7279188
关键词：
2-mercaptoethylguanidine 3-nitrotyrosine Adult Respiratory Distress Syndrome Aftercare Alveolar Macrophages Animal Model Animals Anions Antioxidants Apoptosis Apoptotic Appearance Architecture Arginine Asthma Bacteria Behavior Binding Biochemical Birth Bleomycin Blocking Antibodies Bone Marrow Breathing CD44 gene Cations Caveolae Caveolins Cell Count Cell membrane Cells Cessation of life Chemicals Chemotaxis Co-Immunoprecipitations Collectins Condition Consensus Sequence Copper Cysteine Cytokine Gene Cytoskeleton DNA DNA Sequence Rearrangement Data Diffusion Disease Disruption Effectiveness of Interventions Electrons Enzymes Exposure to Family Fc Receptor Fibrosis GAG Gene Gelatinase A Gene Expression Generations Growth Factor HMMR gene Heat shock proteins Host Defense Hyaluronan Hyaluronic Acid Hyaluronic Acid Binding Hydrogen Peroxide Hydroxide Ion Hydroxides Hyperoxia In Vitro Infection Infiltration Inflammation Inflammatory Inflammatory Infiltrate Inflammatory Response Injury Irrigation Knock-out Knockout Mice Lipid Peroxidation Lipids Literature Localized Lung M40401 Macrophage Activation Measures Mediating Mediator of activation protein Membrane Lipids Membrane Microdomains Mitochondria Modeling Modification Molecular Molecular Weight Mus Nature Nitrates Nitric Oxide Nitric Oxide Synthase Nitrites Nitrogen Nitrogen Dioxide Nucleic Acids Nystatin Oxygen Oxygenases Pattern Peptide antibodies Peptides Peroxidase Peroxides Peroxonitrite Phagocytosis Phospholipids Physiological Pneumonia Positioning Attribute Post-Translational Protein Processing Principal Investigator Production Property Protein Family Protein Isoforms Proteins Pulmonary Surfactant-Associated Protein A Pulmonary Surfactant-Associated Protein D Pulmonary Surfactants Pump Rate Reaction Reactive Nitrogen Species Reactive Oxygen Species Regulation Relative (related person)Reporting Respiratory Burst Respiratory Chain Rodent Role Saline Selenomethionine Side Signal Transduction Site Sodium Stress Superoxide Dismutase Superoxides Surface Surface Tension Testing Therapeutic Intervention Thinking Time Tissues Toxic effect Transgenic Organisms Transition Elements Tyrosine Virus Wild Type Mouse adduct caveolin 1 cell injury cell motility density enzyme activity ethyl nitrite human disease immune function in vivo indium-bleomycin inhibitor/antagonist injured lung injury macrophage member methyl-beta-cyclodextrin molecular size neutrophil nitrate nitration nitrogen tetroxide nitrogen trioxide nitroxyl novel oxidation protein function receptor research study response response to injury scaffold stress protein surfactant

项目摘要

DESCRIPTION (provided by applicant): Oxidative and nitrative stresses are thought to be critical mediators of lung injury and subsequent induction of inflammatory responses. Proteins constitute a major target of superoxide (O) and nitric oxide (NO) reactivity. Under exposure to these reactive stresses, protein modifications such as S-nitrosylation (SNO), carbonylation and 3-nitrotyrosine (3NT) occur and have been shown to act both physiologically and pathologically by altering protein function. The pulmonary collectins, Surfactant Proteins (SP)-A and SP-D modulate immune functions in the lung and nitration of SP-A abrogates its functions. Preliminary data indicate differential localization and expression of SNO and 3NT after bleomycin-induced lung injury in rodents. In addition, we have documented dramatic increases in lavage hyaluronic acid (HA) in injured lungs and HA-binding peptide treatment limits the inflammatory and fibrotic response to injury. Interestingly, reactive oxygen and nitrogen species (RONS) fragment high molecular weight (HMVV) HA into low molecular weight (LMW) forms that promote macrophage activation, cytokine gene expression and chemotaxis. The HA receptors CD44 and RHAMM have been implicated in these inflammatory responses both in vitro and in vivo. Preliminary data indicate that anti-RHAMM antibody blocks SP-A-mediated macrophage chemotaxis and that membrane lipid rafts are required for SP and HA signaling. Using the rodent intratracheal bleomycin model of lung injury, we will test the hypothesis that RONS, occurring as a result of lung injury, generate both GAG adducts and the post.translational modification of specific protein targets that collectively regulate macrophage activation, inflammatory cytokine gene expression and chemotaxis so as to promote pulmonary inflammation. Using both pharmacologic (chemical blockers) and transgenic (SP-ND knockouts) approaches, Aim 1 will determine the contribution of RONS to post-translational modifications of SPND, formation of LMW HA and inflammation after bleomycin injury. In addition, using specific antibody and peptide blockers, Aim 2 will focus on defining the roles of HA, CD44 and RHAMM in the regulation of macrophage functions by native and nitrated SP-ND in vitro. In Aim 3, the mechanisms of altered macrophage function will be further examined by determining the formation of a lipid raft-associated signaling unit that regulates SP-ND and LMW HA-mediated macrophage behavior. Therapeutic intervention with specific blockers of lipid rafts will then be tested for their efficacy in blocking inflammation after lung injury. The experiments described in this proposal will define the molecular mechanisms of pulmonary collectin and LMW HA regulation of macrophage function and will explore novel targets for therapeutic intervention to limit the inflammatory response to lung injury.

描述（由申请人提供）：氧化和硝化应激被认为是肺损伤的关键介质，随后诱导炎症反应。蛋白质构成了超氧化物（O）和一氧化氮（NO）反应性的主要靶标。在暴露于这些反应性应力下，发生蛋白质修饰，例如S-亚硝基化（SNO），羰基化和3-硝基酪氨酸（3NT），并已证明通过改变蛋白质功能在生理和病理上起作用。肺部收集蛋白，表面活性剂蛋白（SP）-A和SP-D调节肺中的免疫功能，SP-A的硝化作用废除了其功能。初步数据表明博来霉素诱导的啮齿动物肺损伤后SNO和3NT的差异性位置和表达。此外，我们还记录了受伤的肺部灌洗透明质酸（HA）的急剧增加，HA结合肽治疗限制了对损伤的炎症和纤维化反应。有趣的是，活性氧和氮种（Rons）片段高分子量（HMVV）HA成低分子量（LMW）形式，促进巨噬细胞活化，细胞因子基因表达和趋化性。 HA受体CD44和RHAMM在体外和体内都与这些炎症反应有关。初步数据表明，抗rHAMM抗体会阻断SP-A介导的巨噬细胞趋化性，并且SP和HA信号传导需要膜脂质筏。使用啮齿动理内博霉素的肺损伤模型，我们将检验以下假设：肺部损伤导致的RON会产生GAG加合物和后。对特定蛋白质靶标的翻译修饰，这些蛋白质集体调节巨噬细胞激活，炎症性细胞因子基因表达和化学症，从而促进plosemation proplymation。使用药理学（化学阻滞剂）和转基因（SP-ND敲除）方法，AIM 1将确定RONS对SPND翻译后修饰的贡献，博来霉素损伤后的LMW HA和炎症的形成和炎症。此外，使用特定的抗体和肽阻滞剂，AIM 2将集中于定义HA，CD44和RHAMM在体外由天然和硝化的SP-ND在巨噬细胞功能中的作用。在AIM 3中，通过确定调节SP-ND和LMW HA介导的巨噬细胞行为的脂质筏相关信号单元的形成，将进一步研究改变巨噬细胞功能的机制。然后，将对脂质筏的特定阻滞剂进行治疗干预，然后测试其在肺损伤后阻断炎症的功效。该提案中描述的实验将定义巨噬细胞功能的肺收集蛋白和LMW HA调节的分子机制，并将探索用于治疗干预的新靶标，以限制对肺损伤的炎症反应。