p47 binding partners in endothelial cell function

描述（由申请人提供）：血管内皮的表型在许多人类血管疾病的演变中发生显著变化，重现了胚胎血管发育期间观察到的变化。在对肿瘤或愈合伤口的血管生成反应期间以及在炎性损伤后的血管恢复期间，可以看到类似于上皮细胞向间充质细胞转化的转换，激活迁移和增殖途径。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力学在功能、生物化学和空间水平上的联系。特别是迁移细胞显示出显著的亚细胞极化的近端信号蛋白，其支配细胞骨架动力学以及存活和增殖途径。值得注意的是，内源性产生的反应性氧化剂已显示集中在迁移内皮细胞的前缘，并且似乎是运动和促有丝分裂信号传导所必需的。这些观察结果表明，内皮细胞氧化酶可以类似地靶向前缘结构，并且这种精确的靶向对于保持氧化剂相关信号的保真度可能是必要的。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实际上是未知的。在之前的资助期间，我们确定了一些主要的NADPH氧化酶适配器，p47 phox的蛋白质结合伙伴，并证明了几个蛋白质-蛋白质相互作用的参与，在指定氧化剂相关的信号传导到特定的信号传导模块。在这个应用程序中，我们建议详细研究这些相互作用在改变内皮细胞表型的作用，使用显微镜，生物化学和功能研究的组合。与公共卫生的相关性：在美国，占主要死亡原因的疾病，其中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。我们建议详细调查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。本申请是R 01-HL 67256的竞争性更新。

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.