Diversity Supplement of R01 grant "How Tetraspanins Regulate Vascular Morphogenesis"

R01 资助的多样性补充“四跨膜蛋白如何调节血管形态发生”

• 批准号: 9796052
• 负责人: XIN A ZHANG
• 金额: $ 3.79万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9796052
• 关键词: Affect; Animal Model; Blood Vessels; Cell Adhesion; Cell Adhesion Molecules; Cell membrane; Cell surface; Cell-Matrix Junction; Development; Disease; Endocytosis; Endothelial Cells; Enzymes; Event; Excision; Gangliosides; Glycosphingolipids; Goals; Grant; Integral Membrane Protein; KAI1 gene; Lead; Mediating; Membrane Lipids; Membrane Microdomains; Metabolic; Molecular; Morphogenesis; Organism; Pathologic; Pathologic Neovascularization; Pathology; Prevention; Recycling; Role; Signal Transduction; Stimulus; blood vessel development; cell motility; insight; neovascularization; novel; prevent; response; therapeutic development; trafficking

Vascular morphogenesis requires proper endothelial cell (EC) adhesion and migration. Transmembrane proteins tetraspanins are abundantly and ubiquitously present in endothelia. Our recent finding indicates that tetraspanin CD82 inhibits neovascularization in response to pathological stimuli. Our study also revealed that this tetraspanin inhibits neovascularization by restraining EC movement, restrains EC movement by confining EC adhesions, and confines EC adhesion by facilitating the endocytosis of cell adhesion molecules (CAMs) and preventing the aggregation of lipid rafts at the plasma membrane. The explicit and complete mechanisms that govern CD82-mediated inhibitions of pathological angiogenesis and EC movement, however, still remain largely unknown at the molecular, cellular, and organism levels. To elucidate how CD82 inhibits neovascularization, we hypothesize that, at the cellular level, CD82 down-regulates the dynamic EC-matrix adhesion, which is needed for proper EC movement. At the molecular level, CD82 reduces the functional cell adhesion proteins at the EC surface by altering the molecular landscape of membrane lipids and subsequently the endocytic machinery of ECs. In this project, we will first determine the mechanism by which CD82 selectively restrains pathological neovascularization. We will identify the CD82 effecter(s) that specifically affects pathological neovascularization, determine if CD82 confines angiogenic signaling that preferentially affects pathological neovascularization, and assess the effect of CD82 on the EC event(s) that mainly affects pathological neovascularization. Secondly, we will determine the mechanism by which CD82 alters the trafficking of cell adhesion molecules by examining the effects of CD82 on their endocytosis, recycling, and exosomal release in ECs. Finally, we will determine how CD82 organizes the membrane microdomains of ECs by assessing the activities of glycosphingolipid-metabolic enzymes upon CD82 removal, the regulatory effects of gangliosides on membrane microdomains, and the roles of gangliosides in CAM trafficking and in CD82- mediated inhibitions of EC movement and pathological angiogenesis. Thus, the goal of this project is to understand how CD82 selectively restrains pathological angiogenesis at the molecular, cellular, and organism levels. From these studies, we will delineate the mechanisms by which tetraspanins regulate vascular morphogenesis, establish a novel paradigm between pathological angiogenesis and membrane microdomain organization, and reveal the signaling axis that governs the crosstalk between EC movement and EC adhesion. From the in-depth mechanistic study, we will develop an integrated understanding of the unique features of CD82, which will ultimately lead to the development of therapeutic mean against pathological angiogenesis.

血管形态发生需要适当的内皮细胞（EC）粘附和迁移。 跨膜蛋白四翼烷蛋白在内皮中大量且普遍存在。我们的 最近的发现表明，四翼胺CD82抑制了新血管形成 病理刺激。我们的研究还表明，这种四跨载质抑制了通过 限制EC运动，通过限制EC粘连来限制EC运动，并限制EC 通过促进细胞粘附分子（CAM）的内吞作用并防止粘附 脂质筏在质膜上的聚集。明确和完整的机制 但是，CD82介导的病理血管生成和EC运动的抑制仍然 在分子，细胞和生物体水平上仍然在很大程度上未知。 为了阐明CD82如何抑制新血管形成，我们假设在细胞水平上， CD82下调了动态的EC-矩阵粘附，这是适当的EC运动所需的。 在分子水平上，CD82通过 改变膜脂质的分子景观，然后改变内吞机器 ECS。在这个项目中，我们将首先确定CD82选择性约束的机制 病理新血管形成。我们将确定特异性影响的CD82效应器 病理新生血管形成，确定CD82是否限制了血管生成信号传导 优先影响病理新血管形成，并评估CD82对EC的影响 主要影响病理新生血管形成的事件。其次，我们将确定 CD82通过检查细胞粘附分子的运输的机制 CD82对ECS中内吞作用，回收和外泌体释放的影响。最后，我们会的 确定CD82如何通过评估活动的活动来组织EC的膜微域 去除CD82后，糖脂果脂脂 - 代谢酶，神经节苷脂的调节作用 在膜微区以及神经节在凸轮运输和CD82-中的作用 介导的EC运动和病理血管生成的抑制作用。因此，这个项目的目标 是要了解CD82如何选择性地约束分子的病理血管生成， 细胞和生物水平。 从这些研究中，我们将描述四叠蛋白调节血管的机制 形态发生，在病理血管生成和膜之间建立一种新的范式 微区组织，并揭示控制EC之间串扰的信号轴 运动和EC粘附。从深入的机械研究中，我们将开发一个综合的 了解CD82的独特特征，这最终将导致发展 针对病理血管生成的治疗平均值。