RTP-801 in Cigarette Smoke-Emphysema

RTP-801在香烟烟雾-肺气肿中的应用

• 批准号: 8064153
• 负责人: Rubin M. Tuder
• 金额: $ 33.21万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8064153
• 关键词: Abbreviations; Address; Adenoviruses; Air; Alveolar; Alveolar Cell; Antioxidants; Apoptosis; Brain; Bronchoalveolar Lavage; Cardiovascular Diseases; Cell Death; Cells; Cellular Stress; Cellular Stress Response; Chronic Obstructive Airway Disease; Cigarette smoke-induced emphysema; Complex; Data; Disease; Erythroid; Fibroblasts; Figs - dietary; Gene Expression; Gene Proteins; Genes; Growth Factor; Human; Hypoxia Inducible Factor; Immunofluorescence Immunologic; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Injury; Intervention; Knock-out; Knockout Mice; Knowledge; Left; Lung; Lung Inflammation; Lung diseases; Maintenance; Manuscripts; Mediating; Mediator of activation protein; Medicine; Molecular; Morbidity - disease rate; Mus; Muscle; NF-kappa B; Natural Immunity; Nature; Nuclear; Oxidative Stress; Oxygen; Paper; Pathogenesis; Patients; Physiological; Process; Proto-Oncogene Proteins c-akt; Pulmonary Emphysema; Role; Screening procedure; Signal Pathway; Signal Transduction; Sirolimus; Small Interfering RNA; Stress; Transduction Gene; Transgenic Mice; Tuberous sclerosis protein complex; Up-Regulation; Virus; Work; acquired immunity; biological adaptation to stress; cell growth; cell injury; cigarette smoke-induced; cigarette smoking; design; dynein light chain; human FRAP1 protein; in vivo; insight; lung injury; mTOR protein; mortality; novel; pollutant; programs; response; sensor; small hairpin RNA; transcription factor

DESCRIPTION (provided by applicant): Our work over the past 10 years has uncovered that the disruption of alveolar structural and molecular maintenance program by cigarette smoke contributes to the pathogenesis of emphysema. The present application is centered on the concept that environmental stresses, such as caused by cigarette smoke (CS) and pollutants activate "rheostat" molecules that govern harmful alveolar septal cell responses involving oxidative stress, apoptosis, and alveolar inflammation. In line with this overall paradigm, we propose mechanistic studies targeting the cellular stress response gene RTP801, which was discovered in a screening of genes upregulated by hypoxia inducible factor-1a (HIF-a) and shown to induce oxidative stress and apoptosis. RTP801 inactivates mTORC1 (for mammalian target for rapamycin complex 1), which controls a central cell signaling pathway that acts as a physiological molecular sensor of environmental stresses. mTORC therefore integrates growth factor signaling, cell growth, and survival adaptation to adverse environmental conditions. We hypothesize that oxidative stress caused by CS activates RTP801 expression, which suppresses mTOR signaling in alveolar cells, leading to alveolar cell inflammation and apoptosis. Upregulation of RTP801 causes oxidative stress-triggered release of dynein-light chain molecule 8 (LC8) from I?B and NF-?B-activation. Our preliminary data demonstrate that RTP801 knockout mice are protected against lung inflammation, apoptosis, and emphysema caused by cigarette smoke. Specific Aim 1 will demonstrate that the stress response gene RTP801 is required for cigarette smoke- induced emphysema in mice. Specific Aim 2 will demonstrate that RTP801-induced lung inflammation and apoptosis by cigarette smoke are mediated by suppression of its downstream target mTOR. Specific Aim3 will demonstrate that RTP801 suppression of mTOR signaling causes oxidative stress-dependent displacement of the dynein-light chain molecule 8 (LC8) from I?B, leading to activation of NF-?B. These aims rely on a translational and comprehensive approach that includes knockout and transgenic mice, in vivo and in vitro manipulation of RTP801/mTOR/AKT gene expression and signaling with adenoassociated virus or adenovirus gene transduction, and siRNA or short hairpin RNA, complementary pharmacological interventions aimed at mTOR and oxidative stress, and detailed morphometric and cellular assessments of alveolar injury. Our proposal advances the concept that CS-mediated lung injury in emphysema is controlled by alveolar septal cells, as the result of the interaction of oxidative stress and molecular switches involved in cell stress responses. This interaction mediated by RTP801 may constitute a critical modulator of innate and acquired immunity and of cellular protection by growth factors, which are central to the pathogenesis of emphysema. PUBLIC HEALTH RELEVANCE: Chronic obstructive pulmonary diseases (COPD) are the 4th leading cause of mortality and morbidity, with worldwide impact. Emphysema is part of COPD and compromises about 20-60% of patients with the disease. In emphysema, the lung literally disappears, leaving behind large holes, a profound inability to breath air into and out of the lungs, and decrease in the ability of the lungs to get oxygen. Our concept is that environmental stresses, such as those cause by cigarette smoke, cause the activation of cellular responses that are originally designed to protect the host. One of these sensor molecules is RTP801. The activation of RTP801 by cigarette smoke activates inflammation and causes lung cell death. Our proposal is to prove that RTP801 is involved in emphysema caused by cigarette smoke, and the potential mechanisms involved in this process. Our studies have relevance to lung diseases, and also to other manifestations of disease caused by cigarette smoke, such as weigh loss due to muscle damage and cardiovascular disease.

描述（由申请人提供）：过去10年中，我们的工作发现，香烟烟雾通过香烟烟雾造成了肺泡结构和分子维持程序的破坏有助于肺气肿的发病机理。本应用集中在以下概念上：环境应力（例如由香烟烟雾（CS）引起的，污染物和污染物都激活了涉及氧化应激，细胞凋亡和肺泡炎症的有害肺泡分子细胞反应的“风湿病”分子。与这一整体范式相一致，我们提出了针对细胞应激反应基因RTP801的机械研究，该研究是在筛选缺氧诱导因子-1a（HIF-A）上调的基因中发现的，并显示出诱导氧化应激和凋亡。 RTP801使MTORC1失活（对于雷帕霉素复合物1的哺乳动物靶标），该型雷帕霉素络合物控制着中央细胞信号通路，该途径充当环境应力的生理分子传感器。因此，MTORC整合了生长因子信号传导，细胞生长和对不利环境条件的生存适应。我们假设由CS引起的氧化应激激活RTP801的表达，这抑制了肺泡细胞中MTOR信号的传导，从而导致肺泡细胞炎症和凋亡。 RTP801的上调导致氧化应激触发的释放，从I？b和nf- b-激活中引起了动力蛋白光链分子8（LC8）。我们的初步数据表明，RTP801基因敲除小鼠受到烟雾烟雾引起的肺部炎症，凋亡和肺气肿的保护。具体的目标1将证明应力反应基因RTP801是小鼠烟雾诱导的肺气肿所必需的。具体的目标2将证明RTP801诱导的肺部炎症和香烟烟雾的凋亡是通过抑制其下游靶MTOR介导的。具体的AIM3将证明RTP801 MTOR信号传导的抑制会导致I？B的Dynein-Light链分子8（LC8）的氧化应激依赖性位移，从而导致NF-？B的激活。 These aims rely on a translational and comprehensive approach that includes knockout and transgenic mice, in vivo and in vitro manipulation of RTP801/mTOR/AKT gene expression and signaling with adenoassociated virus or adenovirus gene transduction, and siRNA or short hairpin RNA, complementary pharmacological interventions aimed at mTOR and oxidative stress, and detailed morphometric and cellular评估肺泡损伤。我们的提议提出了这样一个概念，即CS介导的肺气肿中的肺损伤受肺泡间隔细胞的控制，这是由于氧化应激和与细胞应激反应有关的分子开关的相互作用而导致的。 RTP801介导的这种相互作用可能构成了先天性和获得免疫力的关键调节剂，并通过生长因子对细胞保护，这对于肺气肿的发病机理至关重要。 公共卫生相关性：慢性阻塞性肺部疾病（COPD）是死亡率和发病率的第四个主要原因，具有全球影响。肺气肿是COPD的一部分，约有20-60％的患有该疾病的患者。在肺气肿中，肺实际上消失了，留下了大孔，深刻无法向肺部呼吸空气，并降低了肺部氧气的能力。我们的概念是，环境压力（例如香烟烟雾）会导致最初旨在保护宿主的细胞反应的激活。这些传感器分子之一是RTP801。香烟烟雾激活RTP801会激活炎症并导致肺部细胞死亡。我们的建议是证明RTP801参与了由香烟烟雾引起的肺气肿以及此过程中涉及的潜在机制。我们的研究与肺部疾病有关，也与香烟烟雾引起的其他疾病的表现有关，例如由于肌肉损伤和心血管疾病而引起的称重损失。