Impaired Glycosylation of Vascular Endothelial Growth Factor Receptor 2 in Tumor Angiogenesis

肿瘤血管生成中血管内皮生长因子受体 2 的糖基化受损

• 批准号: 9404509
• 负责人: Kevin Brown Chandler
• 金额: $ 0.03万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9404509
• 关键词: Affect; Angiogenic Switch; BAY 54-9085; Binding Proteins; Blood Vessels; Cell membrane; Cell surface; Clathrin; Development; Drug resistance; Endocytosis; Endosomes; Endothelial Cells; Exposure to; Glycopeptides; Goals; Golgi Apparatus; Growth; Hypoxia; Imaging Techniques; Immunoprecipitation; Impairment; KDR gene; Knowledge; Lead; Ligands; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Membrane; Molecular Biology; Neoplasm Metastasis; Organelles; Pathologic; Pathologic Neovascularization; Pattern; Phosphorylation; Play; Polysaccharides; Population; Process; Protein Glycosylation; Proteins; Proteomics; Receptor Activation; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Recycling; Residencies; Resistance; Resistance development; Role; Signal Transduction; Subcellular Fractions; Tumor Angiogenesis; Tumor Cell Line; UV Radiation Exposure; Vascular Endothelial Growth Factors; Vascularization; Western Blotting; Work; angiogenesis; antiangiogenesis therapy; bevacizumab; cancer therapy; crosslink; extracellular; glycoproteomics; glycosylation; inhibitor/antagonist; link protein; molecular imaging; multicatalytic endopeptidase complex; novel; proteostasis; public health relevance; receptor; screening; sugar; trafficking; tumor; tumor growth

 DESCRIPTION (provided by applicant): Angiogenesis (the development of new blood vessels from established vasculature) is essential for tumor growth and metastasis. Vascular endothelial growth factor receptor 2 (VEGFR-2) is a receptor tyrosine kinase (RTK) involved in angiogenic signaling in normal and pathological conditions. Tumors frequently express high levels of hypoxia-induced VEGF ligands that target VEGFR-2 to support tumor vascularization and growth. While VEGFR-2 inhibitors including bevacizumab, sorafenib and sunitinib are used for cancer treatment, the development of resistance to these agents poses an obstacle to effective management of cancer. Recently, impaired trafficking and ligand-independent VEGFR-2 activation and signaling in the Golgi have been demonstrated. Intriguingly, disruption of N-glycosylation affects VEGFR-2 trafficking and may play a role in this process. However, the impact of VEGFR-2 glycosylation on trafficking and receptor activation is largely unexplored. We hypothesize that altered protein glycosylation in pathological circumstances contributes to the sustained localization of VEGFR-2 to the Golgi apparatus by affecting trafficking of VEGFR-2 and leads to ligand-independent activation and signaling from the Golgi. Therefore, we propose to study the impact of VEGFR-2 glycosylation on VEGFR-2 trafficking and activation. We plan to explore this hypothesis by (A) establishing the glycosylation status of membrane and subcellular fractions of VEGFR-2 and examining their impact on receptor activation, (B) screening for glycosylation-dependent VEGFR-2 protein interactions in cell membrane and Golgi fractions, and (C) investigating the role of VEGFR-2 glycosylation in protein recycling, turnover and degradation. A major goal of this proposal is to explore the role of glycosylation in VEGFR-2 trafficking, signaling and aberrant tumor angiogenesis. I will analyze VEGFR-2 glycopeptides and released N-glycans and screen for VEGFR-2 glycosylation-dependent protein interactions using UV-activated crosslinking sugars via mass spectrometry. Results from this study will illuminate the role of glycosylation in VEGFR-2 trafficking and signaling, including: characterization of differences in glycosylation between Golgi and extracellular fractions, assessment of changes in receptor activation and signaling associated with changes in glycosylation, identification of glycosylation-specific VEGFR-2 interacting partners that modulate VEGFR-2 trafficking, and determination of the impact of receptor glycosylation on protein recycling and degradation.

 描述（由申请人提供）：血管生成（从已建立的血管系统发育出新血管）对肿瘤生长和转移至关重要。血管内皮生长因子受体2（VEGFR-2）是一种受体酪氨酸激酶（RTK），在正常和病理条件下参与血管生成信号。肿瘤经常表达高水平的低氧诱导的VEGF配体，其靶向VEGFR-2以支持肿瘤血管形成和生长。虽然VEGFR-2抑制剂包括贝伐单抗、索拉非尼和舒尼替尼用于癌症治疗，但对这些药物的耐药性的发展对癌症的有效管理构成障碍。最近，受损的运输和配体非依赖性VEGFR-2的激活和信号转导在高尔基体已被证明。有趣的是，N-糖基化的破坏影响VEGFR-2的运输，并可能在这一过程中发挥作用。然而，VEGFR-2糖基化对运输和受体活化的影响在很大程度上尚未探索。我们假设，改变蛋白质糖基化在病理情况下，通过影响VEGFR-2的运输，导致配体非依赖性激活和信号转导的高尔基体的VEGFR-2的持续本地化。因此，我们建议研究VEGFR-2糖基化对VEGFR-2运输和活化的影响。我们计划通过（A）建立VEGFR-2的膜和亚细胞组分的糖基化状态并检查其对受体活化的影响，（B）筛选细胞膜和高尔基体组分中糖基化依赖性VEGFR-2蛋白相互作用，以及（C）研究VEGFR-2糖基化在蛋白质再循环、周转和降解中的作用来探索这一假设。该提案的一个主要目标是探索糖基化在VEGFR-2运输、信号传导和异常肿瘤血管生成中的作用。我将分析VEGFR-2糖肽和释放的N-聚糖，并通过质谱法使用UV活化的交联糖筛选VEGFR-2糖基化依赖性蛋白质相互作用。这项研究的结果将阐明糖基化在VEGFR-2运输和信号传导中的作用，包括：高尔基体和细胞外部分之间糖基化差异的表征，与糖基化变化相关的受体活化和信号传导变化的评估，调节VEGFR-2运输的糖基化特异性VEGFR-2相互作用配偶体的鉴定，以及测定受体糖基化对蛋白质再循环和降解的影响。