C/EBP Beta Regulation of Lung Inflammation

肺部炎症的 C/EBP Beta 调节

• 批准号: 6778762
• 负责人: Arlene A Stecenko
• 金额: $ 38.25万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-12 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6778762
• 关键词: adult respiratory distress syndrome; aerosols; chimeric proteins; disease /disorder model; endotoxins; enhancer binding protein; enzyme linked immunosorbent assay; fibronectins; gel mobility shift assay; gene expression; genetic transcription; immunoregulation; inflammation; interleukin 6; interleukin 8; intravenous administration; lung injury; molecular pathology; nuclear factor kappa beta; protein isoforms; protein structure function; respiratory function; sheep; tumor necrosis factor alpha

DESCRIPTION (provided by applicant): Ordinarily, neutrophilic inflammation is an acute response that either resolves or transitions to a chronic lymphocytic process. However, in several lung diseases including sepsis induced acute lung injury, neutrophil-dominated inflammation persists throughout the course of the disease. Activation of the transcription factor NF-?B is a proximal trigger for neutrophilic inflammation, but factors responsible for termination of the neutrophilic response (or failure to do so) and initiations of the repair process are less clear. In normal human airway epithelial cells and in preliminary experiments in animals, we have evidence that termination of the inflammatory response in the lungs depends on selective increased production of the inhibitory isoform of the transcription factor C/EBP( called p20. The DNA binding sites for C/EBP( and NF-?B are in close proximity in the promoter region of several genes and interactions between the two factors results in synergistic enhancement of gene expression by the activator C/EBP( isoform and inhibition by the dominant negative inhibitory isoform (p20). As both experimental tools and potential therapeutics, we have developed two unique recombinant fusion proteins rendered cell permeable by inclusion of a membrane translocation sequence (MTS). One of these proteins (I?B(((N)-MTS) inhibits activation of NF-?B and the other (p20-MTS) delivers the inhibitor isoform of C/EBP( to cell nuclei. Our goal is to use these tools to elucidate the roles of the two transcription factors, NF-?B and C/EBP(, in the pathogenesis of endotoxin induced lung injury and the early phase of lung repair in a well established sheep model. Specifically, we aim to: 1) in anesthetized sheep, determine the early time course of NF-?B and C/EBP( activity in liver and lung following endotoxemia, relate these changes to measurements of lung function and cellular and biochemical responses, and determine the effects of intravenous delivery of either the inhibitor of NF-?B or of C/EBP( on the response to endotoxemia; 2) in chronically instrumented, unanesthetized sheep, determine effects of intravenous administration of either the inhibitor of NF-?B or the inhibitor of C/EBP( on sub-acute physiologic, cellular and biochemical responses to endotoxin and on the early phase of recovery of the lungs from endotoxin-induced injury; 3) in chronically instrumented, unanesthetized sheep, determine effects of aerosol administration of either inhibitor on the response to endotoxin and on the early phase of recovery of the lungs from injury; and 4) from the above studies, select the most promising therapeutic regimen and determine effects of its administration four hours after endotoxemia on the subsequent course of the endotoxin response in chronically instrumented unanesthetized sheep.

描述（由申请人提供）：通常，中性粒细胞炎症是一种急性反应，要么消退，要么转变为慢性淋巴细胞过程。 然而，在包括败血症引起的急性肺损伤在内的几种肺部疾病中，中性粒细胞主导的炎症在整个疾病过程中持续存在。 转录因子 NF-κB 的激活是中性粒细胞炎症的近端触发因素，但导致中性粒细胞反应终止（或未能终止）和修复过程启动的因素尚不清楚。 在正常人气道上皮细胞和动物初步实验中，我们有证据表明，肺部炎症反应的终止取决于选择性增加转录因子 C/EBP（称为 p20）抑制性亚型的产生。C/EBP（和 NF-κB 的 DNA 结合位点在多个基因的启动子区域非常接近，两个因子之间的相互作用导致协同作用 通过激活剂 C/EBP( 同工型) 增强基因表达，并通过显性失活抑制同工型 (p20) 进行抑制。作为实验工具和潜在的治疗方法，我们开发了两种独特的重组融合蛋白，通过包含膜易位序列 (MTS) 使细胞具有通透性。其中一种蛋白 (I?B((N)-MTS) 抑制 NF-κB 和另一种 (p20-MTS) 将 C/EBP( 的抑制剂亚型) 递送至细胞核。我们的目标是使用这些工具来阐明 NF-κB 和 C/EBP( 两种转录因子在内毒素诱导的肺损伤的发病机制中以及在完善的绵羊模型中肺修复的早期阶段中的作用。具体来说，我们的目标是： 1) 在麻醉中 确定内毒素血症后 NF-κB 和 C/EBP（肝和肺活动）的早期过程，将这些变化与肺功能、细胞和生化反应的测量结果联系起来，并确定静脉注射 NF-κB 或 C/EBP 抑制剂（对内毒素血症反应的影响；2）在长期使用仪器、未麻醉的绵羊中，确定 静脉注射 NF-κB 抑制剂或 C/EBP 抑制剂（对内毒素的亚急性生理、细胞和生化反应以及内毒素引起的肺部损伤恢复的早期阶段；3）在长期使用仪器、未麻醉的绵羊中，确定任一抑制剂的气雾给药对内毒素反应和早期阶段的影响 肺部从损伤中恢复； 4) 从上述研究中，选择最有希望的治疗方案，并确定内毒素血症后四小时给药对长期使用仪器的未麻醉绵羊的内毒素反应的后续过程的影响。