p47 binding partners in endothelial cell function

描述(申请人提供)：血管内皮细胞的表型在许多人类血管疾病的演变过程中发生了巨大的变化，概括了胚胎血管发育过程中的变化。一种类似于上皮细胞向间充质细胞转变的开关，激活迁移和增殖途径，在对肿瘤或愈合伤口的血管生成反应中，也在炎症损伤后血管的重建过程中看到。内皮细胞迁移和增殖的这种耦合反映了细胞命运决定和细胞骨架机制在功能、生化和空间水平上的联系。尤其是迁移细胞，其近端信号蛋白表现出显著的亚细胞极化，这些信号蛋白控制着细胞骨架的动态以及生存和增殖途径。值得注意的是，内源性产生的活性氧化剂已被证明集中在迁移的内皮细胞的前沿，似乎对运动和有丝分裂信号都是必需的。这些观察表明，内皮细胞氧化酶可能同样针对前沿结构，这种精确的靶向可能对于保持氧化剂相关信号的保真度至关重要。然而，这种假定的亚细胞氧化酶定位的分子基础和生物学原理几乎是未知的。在之前的资助期间，我们确定了主要NADPH氧化酶接头p47Phox的一些蛋白质结合伙伴，并证明了几种蛋白质-蛋白质相互作用参与了特定信号模块的氧化剂相关信号的指定。在这项应用中，我们建议结合显微镜、生化和功能研究，详细研究这些相互作用在改变内皮细胞表型中的作用。与公共卫生的相关性：在美国，占主要死亡原因的疾病，包括癌症和心血管疾病，涉及血管内皮细胞表型的根本变化。我们建议详细研究这些变化的生化基础的一个方面，希望逆转或防止这些变化。此应用程序是R01-HL67256的竞争性续订。

DESCRIPTION (provided by applicant): The phenotype of the vascular endothelium changes dramatically in the evolution of a number of human vascular diseases, recapitulating changes seen during embryonic vascular development. A switch similar to the epithelial-to-mesenchymal transition, activating migration and proliferation pathways, is seen during the angiogenic response to tumors or healing wounds and also during the restitution of vessels following inflammatory injury. This coupling of endothelial cell migration and proliferation reflects the linkage between cell fate decisions and cytoskeletal mechanics at functional, biochemical, and spatial levels. Migrating cells in particular display striking subcellular polarization of proximal signaling proteins which govern cytoskeletal dynamics as well as survival and proliferation pathways. Notably, endogenously- produced reactive oxidants have been shown to concentrate at the leading edge of migrating endothelial cells, and appear to be necessary for both locomotion and mitogenic signaling. These observations suggest that the endothelial cell oxidase may be similarly targeted to leading edge structures, and that such precise targeting may be essential to preserve the fidelity of oxidant-related signals. However, the molecular basis and biological rationale for such putative subcellular oxidase localization is virtually unknown. During the prior funding period, we identified a number of protein binding partners of the principal NADPH oxidase adapter, p47phox, and demonstrated the involvement of several protein-protein interactions in specifying oxidant-related signaling to specific signaling modules. In this application, we propose to examine in detail the role of these interactions in changing the endothelial phenotype, using a combination of microscopic, biochemical, and functional studies. Relevance to Public Health: Diseases that account for the leading causes of death in the United States, among them cancer and cardiovascular disease, involve fundamental changes in the phenotype of the vascular endothelium. We propose to investigate in detail one facet of the biochemical basis for these changes, in hopes of reversing or preventing these changes. This application is a competing renewal of R01-HL67256.