C/EBP Beta Regulation of Lung Inflammation

肺部炎症的 C/EBP Beta 调节

• 批准号: 7228878
• 负责人: Arlene A Stecenko
• 金额: $ 35.96万
• 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/7228878
• 关键词: Acute; Acute Lung Injury; Aerosols; Affect; Alveolar Macrophages; Amino Acids; Anesthesia procedures; Animals; Binding Sites; Biochemical; Blood Vessels; Body Fluids; Bronchoalveolar Lavage Fluid; CCAAT-Enhancer-Binding Protein-beta; CCAAT-Enhancer-Binding Proteins; Cell Nucleus; Cells; Cessation of life; Chest; Chimeric Proteins; Chronic; Clinical; Collection; DNA Binding; Disease; Dominant-Negative Mutation; Dose; Elevation; Endotoxemia; Endotoxins; Epithelial Cells; Event; Failure; Gene Expression; Generations; Genes; Goals; Hour; Human; I Kappa B-Alpha; IL8 gene; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-6; Intervention; Intravenous; Investigation; Liver; Localized; Lung; Lung Inflammation; Lung diseases; Lymphocyte; Measurement; Mediating; Membrane; Methods; Modeling; NF-kappa B; Onset of illness; Operative Surgical Procedures; Organ; Pathogenesis; Phase; Physiological; Plasmid Cloning Vector; Play; Pneumonia; Preparation; Process; Production; Promoter Regions; Protein Isoforms; Proteins; Recombinant Fusion Proteins; Recovery; Regulation; Relative (related person); Respiratory physiology; Role; Route; Sepsis; Sheep; Stimulus; Structure; Structure of parenchyma of lung; Testing; Therapeutic; Time; Tissue Banks; Tissues; Transgenes; Treatment Protocols; Wound Healing; base; day; design; factor C; gene interaction; in vivo; inhibitor/antagonist; instrument; intravenous administration; lung injury; mortality; neutrophil; protein expression; repaired; research study; response; tool; transcription factor; vector

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(申请人提供)：通常，中性粒细胞炎症是一种急性反应，可分解或转变为慢性淋巴细胞过程。然而，在包括败血症所致的急性肺损伤在内的几种肺部疾病中，以中性粒细胞为主的炎症在整个病程中持续存在。转录因子核因子-βB的激活是中性粒细胞炎症的近端触发因素，但负责终止(或未能终止)中性粒细胞反应和修复过程的因素尚不清楚。在正常的人呼吸道上皮细胞和动物的初步实验中，我们有证据表明，肺部炎症反应的终止依赖于转录因子C/EBP抑制亚型(称为p20)选择性地增加。C/EBP(和核因子-βB)的DNA结合部位位于多个基因的启动子区域，两者之间的相互作用导致激活因子C/EBP(异构体)协同促进基因表达，并通过显性负抑制异构体(P20)抑制基因表达。作为实验工具和潜在的治疗手段，我们开发了两种独特的重组融合蛋白，通过包含膜转位序列(MTS)使细胞可渗透。其中一种蛋白(I？B(N)-MTS))抑制核因子-？B的激活，另一种(p20-MTS)将C/EBP的抑制亚型运送到细胞核。我们的目标是利用这些工具来阐明两种转录因子--核因子-β和C/EBP在内毒素诱导的绵羊肺损伤和早期肺修复中的作用。具体地说，我们的目标是：1)在麻醉绵羊中，测定内毒素血症后肝脏和肺组织中核因子-βB和C/EBP(活性)的早期时间进程，并将这些变化与肺功能和细胞和生化反应的测量联系起来，并确定静脉注射核因子-βB抑制剂或C/EBP(对内毒素血症反应的影响；2)在慢性仪器化的非麻醉绵羊中，确定静脉注射核因子-βB抑制剂或C/EBP抑制剂的效果(对内毒素的亚急性生理、细胞和生化反应以及内毒素诱导的肺损伤恢复的早期阶段)；3)在慢性仪器化的非麻醉绵羊中，确定雾化给药对内毒素的反应和肺损伤恢复的早期的影响；以及4)从上述研究中，选择最有希望的治疗方案，并确定其在内毒素血症后4小时给药对内毒素反应后续过程的影响。