How Tetraspanins Regulate Vascular Morphogenesis

四跨膜蛋白如何调节血管形态发生

• 批准号: 9362663
• 负责人: XIN A ZHANG
• 金额: $ 37万
• 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/9362663
• 关键词: 1-Phosphatidylinositol 4-Kinase; Ablation; Adhesions; Affect; Angiogenic Factor; Animal Model; Biochemical; Biophysics; Blood Vessels; C-terminal; CD44 gene; CD81 gene; Caveolae; Cell Adhesion; Cell Adhesion Molecules; Cell membrane; Cell surface; Cell-Matrix Junction; Clathrin; Cysteine; Cytoplasmic Tail; Cytosol; Development; Disease; Endocytosis; Endothelial Cells; Enzymes; Event; Excision; Extracellular Matrix; Gangliosides; Glycosphingolipids; Goals; Gray unit of radiation dose; Growth Factor Receptors; Heterogeneity; Integral Membrane Protein; Integrins; Intracellular Signaling Proteins; KAI1 gene; Lead; Lipids; Mediating; Membrane; Membrane Lipids; Membrane Microdomains; Membrane Proteins; Metabolic; Modeling; Molecular; Morphogenesis; Mus; Organism; Pathologic; Pathologic Neovascularization; Pathology; Pathway interactions; Prevention; Proteins; Recycling; Role; Signal Transduction; Sorting - Cell Movement; Stimulus; Structure; Surface; Testing; Transmembrane Domain; angiogenesis; blood vessel development; cell motility; disulfide bond; extracellular; insight; migration; neovascularization; novel; postnatal; 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)的黏附和迁移。 跨膜蛋白Tetraspanins广泛存在于血管内皮细胞中。我们的 最近的发现表明，Tetraspanin CD82抑制新生血管的形成 病理性刺激。我们的研究还显示，这种四环素通过以下方式抑制新生血管 抑制EC运动，通过限制EC粘连来抑制EC运动，限制EC 通过促进细胞黏附分子(CAM)的内吞作用和防止 脂筏聚集在质膜上。明确和完整的机制， 然而，控制CD82介导的病理性血管生成和EC运动的抑制 在分子、细胞和生物体水平上仍然很大程度上未知。 为了阐明CD82如何抑制新生血管，我们假设，在细胞水平上， CD82下调EC-基质的动态黏附，这是EC正常运动所必需的。 在分子水平上，CD82通过以下途径减少EC表面的功能性细胞黏附蛋白 改变膜脂的分子格局，进而改变血管内皮细胞的内吞机制 ECS。在这个项目中，我们将首先确定CD82选择性抑制的机制 病理性新生血管。我们将确定CD82效应器(S)，它特定地影响 病理性新生血管，确定CD82是否限制血管生成信号 优先影响病理性新生血管，并评估CD82对EC的影响 事件(S)，主要影响病理性新生血管。其次，我们将确定 CD82通过检测细胞黏附分子改变细胞黏附分子运输的机制 CD82对内皮细胞内吞、循环和胞外释放的影响。最后，我们会 通过评估CD82的活性来确定CD82如何组织内皮细胞的膜微域 神经节苷脂对CD82清除的调节作用 膜微区，以及神经节苷脂在CAM转运和CD82-中的作用 介导内皮细胞运动和病理性血管生成的抑制。因此，这个项目的目标是 是为了了解CD82如何从分子上选择性地抑制病理性血管生成， 细胞水平和生物体水平。 从这些研究中，我们将描述河豚毒素调节血管的机制。 形态发生，建立病理性血管生成和膜之间的新范式 微域组织，并揭示控制EC之间串扰的信号轴 运动和EC粘附性。从深入的机理研究出发，我们将开发出一套完整的 了解CD82的独特功能，这最终将导致 针对病理性血管生成的治疗手段。