C/EBP Beta Regulation of Lung Inflammation

描述（由申请方提供）：通常，嗜酸性炎症是一种急性反应，可消退或转变为慢性淋巴细胞过程。 然而，在包括脓毒症引起的急性肺损伤在内的几种肺部疾病中，嗜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-？B是嗜酸性炎症的近端触发因子，但导致嗜酸性反应终止（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的初步实验中，我们有证据表明，肺中炎症反应的终止取决于转录因子C/EBP（称为p20）的抑制性同种型的选择性增加。 C/EBP（和NF-？B在几个基因的启动子区域中非常接近，并且两个因子之间的相互作用导致激活剂C/EBP（同种型）协同增强基因表达和显性负抑制同种型（p20）的抑制。 作为实验工具和潜在的治疗剂，我们已经开发了两种独特的重组融合蛋白，通过包含膜易位序列（MTS）使细胞具有渗透性。 其中一种蛋白质（I？B（N）-MTS））抑制NF-？B和另一个（p20-MTS）将C/EBP的抑制剂同种型递送至细胞核。 我们的目标是使用这些工具来阐明两个转录因子，NF-？B和C/EBP（）在绵羊内毒素性肺损伤的发病机制和肺修复的早期阶段。 具体而言，我们的目标是：1）在麻醉羊，确定NF-？B和C/EBP（内毒素血症后肝和肺中的活性，将这些变化与肺功能和细胞及生化反应的测量相关，并确定静脉内递送NF-？B或C/EBP（对内毒素血症的反应; 2）在慢性仪器，未麻醉的羊，确定静脉给药的NF-？B或C/EBP抑制剂（对内毒素的亚急性生理、细胞和生物化学反应，以及对肺从内毒素诱导的损伤中恢复的早期阶段; 3）在慢性仪器化的、未麻醉的绵羊中，确定气雾剂施用任一抑制剂对内毒素反应和对肺从损伤中恢复的早期阶段的影响;和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4小时给药对慢性仪器化未麻醉绵羊中内毒素反应的后续过程的影响。

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.