NO-Modified Biomolecules and Pulmonary Signaling

描述（由申请人提供）：一氧化氮（NO）在肺生理中发挥的核心作用被强调为它发挥的功能的数量，包括维持气道张力，血管张力，炎症，甚至肺的生长和发育。除了这些重要的生理作用外，NO还与许多肺部疾病有关，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产生如此广泛信号的分子机制尚不清楚，此外，NO代谢的破坏如何在病理中发挥作用也不清楚。在炎症中，NO的最重要来源是酶iNOS的诱导形式。inos衍生的NO的一个关键下游效应是硫醇残基的s -亚硝基化生成s -亚硝基硫醇（SNO）。我们假设，通过SNO修饰不同的靶蛋白，inos衍生的NO可以调节促炎和损伤的消退反应。我们已经构建了一个模型，其中巨噬细胞内炎症反应早期产生的NO可用于s -亚硝基酸细胞外靶标，如表面活性剂蛋白d (SP-D)；而在反应后期，随着NO通量的增加和其他氧化剂的产生，细胞内的s -亚硝基化目标，如NF-？B，促进解决和修复。我们计划研究在常驻和招募的巨噬细胞中iNOS和sno降解酶GSNOR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，因为它同时具有炎症和消退/修复阶段，因此是检验我们假设的理想选择。在第一个目标中，这些酶的差异表达将通过过继转移来平衡s -亚硝基化反应。我们将在分子、细胞和器官功能水平上确定巨噬细胞中iNOS和GSNOR缺失的影响。在第二个目标中，我们将研究如何利用特定SNO靶蛋白SP-D的信号机制知识来加重或加剧博莱霉素介导的肺损伤。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。

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.