NO-Modified Biomolecules and Pulmonary Signaling

NO 修饰的生物分子和肺部信号传导

• 批准号: 8707538
• 负责人: Andrew J Gow
• 金额: $ 33.65万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-11 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707538
• 关键词: Adoptive Transfer; Adult Respiratory Distress Syndrome; Alveolar; Alveolar Macrophages; Asthma; Attenuated; Biological; Bleomycin; Blood Vessels; Breathing; Cells; Chronic; Cystic Fibrosis; Data; Enzymes; Epithelial; Epithelium; Equilibrium; Fibrosis; Funding; Generations; Genes; Glutathione; Growth and Development function; Heterotrimeric GTP-Binding Proteins; Inflammation; Inflammatory; Inflammatory Response; Injury; Knock-out; Knowledge; Laboratories; Lung; Lung diseases; Lymphocyte; Measures; Mediating; Metabolism; Modeling; Modification; Molecular; Molecular Weight; Mus; NOS2A gene; Nitric Oxide; Nitric Oxide Synthase; Organ; Outcome; Oxidants; Oxidoreductase; Pathogenesis; Pathology; Patients; Phase; Phenotype; Physiological; Physiology; Play; Pneumonia; Process; Protein Isoforms; Proteins; Proteome; Pulmonary Pathology; Pulmonary Surfactant-Associated Protein D; Recruitment Activity; Resolution; Respiratory System; Role; S-Nitrosoglutathione; S-Nitrosothiols; Signal Transduction; Signaling Molecule; Signaling Protein; Source; Sulfhydryl Compounds; Surface; Tail; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Xenobiotics; design; extracellular; genetic regulatory protein; human NOS2A protein; inflammatory marker; inhaled nitric oxide; inhibitor/antagonist; lung injury; macrophage; novel; persistent pulmonary hypertension; public health relevance; pulmonary function; repaired; response; response to injury; therapeutic target

DESCRIPTION (provided by applicant): The central role that Nitric Oxide (NO) plays within pulmonary physiology is highlighted by the number of functions in which it plays a role including the maintenace of ariway tone, blood vessel tone, inflammation, and even lung growth and development. In addition to these important physiological roles, NO has also been implicated in a number of pulmonary diseases including ARDS, Asthma, and cystic fibrosis. As yet the molecular mechanisms by which this simple diatomic molecule can produce such a wide range of signals is unclear, furthermore, it is unclear how disruption of NO metabolism may play a role in pathology. In inflammation, the most important source of NO is the inducible form of the enzyme iNOS. A key downstream effect of iNOS-derived NO is S-nitrosylation of thiol residues to form S-nitrosothiol (SNO). We hypothesize that, by SNO modification of different target proteins, iNOS-derived NO can regulate both the pro-inflammatory and the resolution responses to injury. We have constructed a model in which NO produced early in the inflammatory response within resident macrophages serves to S-nitrosylate extracellular targets, such as Surfactant Protein-D (SP-D); while later in the response, with increasing fluxes of NO and the generation of other oxidants, intracellular S-nitrosylation of targets, such as NF-?B, promotes resolution and repair. We plan to investigate how the presence of iNOS and the SNO-degrading enzyme, GSNOR, in resident and recruited macrophages alters the outcome of bleomycin-mediated lung injury. We have chosen this injury model as it has both an inflammatory and a resolution/repair phase and is therefore ideal for examining our hypothesis. In the first aim differential expression of these enzymes that balance the S-nitrosylation response will be achieved with the use of adoptive transfer. We will determine the effects of loss of iNOS and GSNOR within resident and recruited macrophages at the molecular, cellular, and organ function level. In the second aim, we will examine how we can use knowledge of the signaling mechanisms of a particular SNO target protein, SP-D, to either accentuate or exacerbate bleomycin-mediated lung injury. These studies use state of the art techniques to determine how NO can signal through S-nitrosylation of different target proteins and may provide novel avenues for therapeutic design.

描述（由申请人提供）：一氧化氮（NO）在肺生理学中发挥的中心作用通过其发挥作用的功能的数量来强调，包括气道张力、血管张力、炎症的维持，甚至肺的生长和发育。除了这些重要的生理作用之外，NO还涉及许多肺部疾病，包括ARDS、哮喘和囊性纤维化。到目前为止，这种简单的双原子分子可以产生如此广泛的信号的分子机制尚不清楚，此外，还不清楚NO代谢的破坏如何在病理学中发挥作用。在炎症中，NO的最重要来源是酶iNOS的诱导形式。iNOS衍生的NO的关键下游效应是硫醇残基的S-亚硝基化以形成S-亚硝基硫醇（SNO）。我们推测，通过SNO修饰不同的靶蛋白，iNOS衍生的NO可以调节促炎反应和对损伤的消退反应。我们已经构建了一个模型，在该模型中，NO产生早期的炎症反应内居民巨噬细胞的S-亚硝基化细胞外的目标，如表面活性蛋白-D（SP-D），而后来的反应，随着流量的增加NO和其他氧化剂的产生，细胞内的S-亚硝基化的目标，如NF-？B，促进消退和修复。我们计划研究iNOS和SNO降解酶GSNOR在巨噬细胞中的存在如何改变博莱霉素介导的肺损伤的结果。我们选择了这种损伤模型，因为它具有炎症和消退/修复阶段，因此是检验我们假设的理想模型。在第一个目标中，平衡S-亚硝基化反应的这些酶的差异表达将通过使用过继转移来实现。我们将在分子、细胞和器官功能水平上确定在驻留和募集的巨噬细胞内iNOS和GSNOR的损失的影响。在第二个目标中，我们将研究如何利用特定SNO靶蛋白SP-D的信号传导机制的知识来加重或加剧博来霉素介导的肺损伤。这些研究使用最先进的技术来确定NO如何通过不同靶蛋白的S-亚硝基化发出信号，并可能为治疗设计提供新的途径。