Redox-Derived Pulmonary Anti-Inflammatory Mediators

描述（由申请人提供）：拟议的研究计划将由一个对开发和测试治疗气道炎症和哮喘的新药策略感兴趣的基础和医生科学家团队进行。初步数据显示，硝基（RNO 2）和α，<$-不饱和酮（R=O）亲电脂肪酸氧化产物（EFOX）是由炎症激活的巨噬细胞产生的，并以增加的浓度存在于哮喘患者支气管肺泡灌洗获得的气道衬里液中。EFOX主要以蛋白加合状态存在（解释了为什么这些物种是最近才发现的），并被视为生物体对炎症的适应性反应的组成部分。研究人员已经观察到EFOX信号通过两种受体依赖性机制（作为有效的PPAR？配体）和通过亲电响应转录因子调节机制。我们打算在a）培养的气道上皮细胞和炎性细胞，B）哮喘小鼠模型和c）从哮喘患者获得的痰液和灌洗液样本中表征与气道炎症相关的条件下EFOX产生的模式和机制。这一奋进将采用一个公正的HPLC-质谱为基础的“亲电捕鱼”的战略。在了解更多关于气道炎症中特定EFOX的结构特征、浓度和分子靶点的信息后，我们将通过体内给予结构相同的合成化合物来评估EFOX在哮喘小鼠模型中的治疗潜力。研究策略旨在验证以下假设：不饱和脂肪酸的亲电硝基和酮衍生物由哮喘气道的氧化炎症环境形成，并介导抗炎细胞信号传导反应。这一奋进具有重要意义，因为它a）提供了对导致哮喘中一类新的炎症调节物质产生的基本分子和细胞机制的关键见解，以及B）测试了用于治疗哮喘患者的有前途的新的基于内源性介体的治疗方法，该方法适合作为临床治疗策略快速部署。

DESCRIPTION (provided by applicant): The proposed research plan will be pursued by a basic and physician scientist team interested in the development and testing of new drug strategies to treat airway inflammation and asthma. Preliminary data reveal that nitro (RNO2) and a,¿-unsaturated keto (R=O) electrophilic fatty acid oxidation products (EFOX) are produced by inflammatory-activated macrophages and are present at increased concentrations in the airway lining fluids obtained by bronchoalveolar lavage of asthmatic patients. EFOXs are found in predominantly protein-adducted states (explaining why these species were just recently discovered), and are viewed as components of the adaptive response of organisms to inflammation. The investigators have observed that EFOX signal via both receptor-dependent mechanisms (as potent PPAR? ligands) and via electrophile-responsive transcription factor-regulated mechanisms. We intend to characterize the patterns and mechanisms of EFOX production during conditions relevant to airway inflammation in a) cultured airway epithelial cells and inflammatory cells, b) a murine model of asthma and c) sputum and lavage samples obtained from asthmatic patients. This endeavor will employ an unbiased HPLC-mass spectrometry-based "electrophile fishing" strategy. After learning more about the structural characteristics, concentrations and molecular targets of specific EFOXs in airway inflammation, we will evaluate the therapeutic potential of EFOXs in a murine model of asthma by in vivo administration of structurally identical synthetic compounds. The research strategy was designed to test the hypothesis that electrophilic nitro and keto derivatives of unsaturated fatty acids are formed by the oxidative inflammatory milieu of asthmatic airways and mediate anti-inflammatory cell signaling responses. This endeavor is of significance because it a) lends critical insight into fundamental molecular and cellular mechanisms leading to the generation of a new class of inflammatory-regulating species in asthma and b) tests a promising new endogenous mediator-based therapeutic approach for treating asthma patients that is amenable for rapid deployment as a clinical therapeutic strategy.