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（应用程序提供）：血管生成（已建立脉管系统的新血管发展）对于肿瘤生长和转移至关重要。血管内皮生长因子受体2（VEGFR-2）是在正常和病理条件下参与血管生成信号传导的受体酪氨酸激酶（RTK）。肿瘤经常表达高水平的缺氧诱导的VEGF配体，这些配体靶向VEGFR-2以支持肿瘤血管形成和生长。虽然包括贝伐单抗，索拉非尼和舒尼尼在内的VEGFR-2抑制剂用于癌症治疗，但对这些药物的抗性发展却是有效治疗癌症的障碍。最近，已证明了高尔基体中的运输和非配体无关的VEGFR-2激活和信号传导。有趣的是，N-糖基化的破坏会影响VEGFR-2运输，并可能在此过程中发挥作用。然而，我们假设在病理情况下改变蛋白质糖基化的影响会导致VEGFR-2通过影响VEGFR-2的运输并导致与配体无依赖性的激活和Golgi信号传递，从而有助于将VEGFR-2持续定位到高尔基体中。因此，我们建议研究VEGFR-2糖基化对VEGFR-2运输和激活的影响。 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退化。该提案的主要目的是探索糖基化在VEGFR-2运输，信号传导和异常肿瘤血管生成中的作用。我将通过质谱法分析VEGFR-2糖肽并释放N-聚糖和筛选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：表征：表征高尔基体和细胞外部分之间糖基化差异的差异，评估受体激活的变化以及与糖基化的变化相关的信号传导的变化，与糖基化的变化相关，糖基化的脉络脉络和素质范围的范围均可互动范围和素质范围的范围，并构成了2-2相互作用的范围，即彼此的互动范围彼此综合综合综合综合群体的综合部分，受体糖基化在蛋白质回收和降解上。