Vascular endothelial growth factor induced lung edema

描述(申请人提供)：成人呼吸窘迫综合征(ARDS) 每年影响美国15万人，死亡率为40%。 目前的治疗只是支持性的。新的治疗方法将需要详细的 对这种疾病的分子基础的理解。世界上最早的 ARDS的表现是发展为非心源性肺水肿。我们 做出了一项新的观察，即肺水肿是由 血管内皮生长因子基因在肺组织中的过表达 使用改良腺病毒载体方法。基因转移与过度表达 经小鼠肺抽提物Northern分析和酶联免疫吸附试验证实。 在肺组织学中观察水肿，并用肺湿重/干重来量化。 伊文氏蓝法测定肺毛细血管对大分子的通透性 染色化验和131L-白蛋白肺漏。这项提议将充分体现出 血管内皮生长因子致肺水肿的内皮机制 通透性，肺泡功能障碍，血管内皮生长因子受体(S)特异性，信号 介导水肿的转导通路和下游效应器。 超微结构信息将通过电子显微镜获得 肺组织中血管内皮生长因子过度表达。血管内皮细胞生长因子亚型的结构与功能关系 肺水肿的发展将使用包含以下内容的Ad载体来定义 各种血管内皮生长因子亚型以及突变型血管内皮生长因子的基因 与特定的血管内皮生长因子受体结合的特异性。过度表达， 免疫抑制和药理抑制剂研究将确定 血管内皮生长因子下游的潜在介质，如内皮一氧化氮 合成酶和一氧化氮以及细胞内信号通路 在血管内皮生长因子诱导的水肿过程中被激活。评估其潜在作用的研究 活化的中性粒细胞在急性肺损伤中作为体内血管内皮生长因子的来源 被开发出来。将测定支气管肺泡灌洗液和血浆中血管内皮生长因子的水平 在急性肺损伤患者中发展一种假说 肺组织中血管内皮生长因子的过度表达可能是促进肺组织血管内皮生长因子升高的机制之一 渗透性。

DESCRIPTION (provided by applicant): Adult respiratory distress syndrome (ARDS) affects 150,000 individuals in the US annually with a mortality of 40 percent. Current treatment is supportive only. New therapies will require a detailed understanding of the molecular basis for this disorder. One of the earliest manifestations of ARDS is the development of noncardiogenic pulmonary edema. We have made the novel observation that pulmonary edema is induced by overexpression of the vascular endothelial growth factor (VEGF) gene in lung using a modified adenovirus vector approach. Gene transfer and overexpression of VEGF were confirmed by Northern analysis of mouse lung extracts and ELISA. Edema was observed in lung histology and quantified by lung wet-to-dry weight ratio and pulmonary capillary permeability to macromolecules by the Evan's blue dye assay and 131 l-albumin lung leak. This proposal will fully characterize the mechanism of VEGF-induced pulmonary edema with regard to endothelial permeability, alveolar dysfunction, VEGF receptor(s) specificity, signal transduction pathways and downstream effectors that mediate the edema. Ultrastructual information will be obtained by electron microscopy of VEGF-overexpressing lungs. Structure-function relationships of VEGF isoforms to the development of pulmonary edema will be defined using Ad vectors containing the genes for the various VEGF isoforms as well as mutant VEGFs which have specificity for binding to a particular VEGF receptor. Overexpression, immunoinhibition and pharmacologic inhibitor studies will define the roles of potential mediators downstream from VEGF such as endothelial nitric oxide synthase and nitric oxide and the intracellular signaling pathways that are activated during VEGF-induced edema. Studies to assess the potential role of activated neutrophils as a source of VEGF in vivo during acute lung injury will be developed. Bronchoalveolar lavage and plasma levels of VEGF will be measured in patients with acute lung injury to develop the hypothesis that overexpression of VEGF in lung may be one mechanism favoring increased permeability.