Endothelial cell decisions integrate soluble and matrix-bound VEGF gradients

内皮细胞决策整合了可溶性和基质结合的 VEGF 梯度

• 批准号: 7511209
• 负责人: Feilim C Mac Gabhann
• 金额: $ 8.27万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7511209
• 关键词: Adherence; Age related macular degeneration; Arteries; Arts; Binding; Blood Vessels; Cell Proliferation; Cells; Clinical Trials; Computer Simulation; Coronary Arteriosclerosis; Cultured Cells; Cytoskeleton; Decision Making; Decision Modeling; Diabetic Retinopathy; Diffuse; Disease; Embryonic Development; Endothelial Cells; Environment; Equilibrium; Exercise; Experimental Designs; Extracellular Matrix; Family; Fibronectins; Frequencies; Genes; Hypoxia; In Vitro; Integrins; Laboratories; Left; Ligand Binding Domain; Ligands; Macular degeneration; Malignant Neoplasms; Masks; Methods; Modeling; Morbidity - disease rate; Mus; Muscle Cells; Mutation; Pathogenesis; Pathway interactions; Pattern; Peripheral; Process; Proliferating; Protein Isoforms; Proteins; Protocols documentation; Public Health; Receptor Activation; Receptor Gene; Receptor Protein-Tyrosine Kinases; Regulation; Relative (related person); Reproduction; Research; Research Personnel; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Stimulus; Techniques; Testing; Therapeutic; Tissues; Training; Transcriptional Activation; Transfection; United States Food and Drug Administration; Up-Regulation; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factors; angiogenesis; base; cell motility; cytokine; density; design; extracellular; genetic manipulation; in vitro Assay; in vivo; inhibitor/antagonist; migration; mortality; mutant; neovascularization; novel; programs; receptor; research study; response; therapeutic target

DESCRIPTION (provided by applicant): Project Summary: The vascular endothelial growth factor (VEGF) family consists of critical cytokines that are implicated in controlling vascular survival and angiogenic sprouting. Recently VEGF inhibitors have been approved by the FDA to treat diseases of hypervascularization (cancer and macular degeneration), while increasing VEGF expression is a major research avenue for ischemic diseases such as peripheral artery disease and coronary artery disease, though clinical trials are as yet unsuccessful. VEGF is involved in at least 70 diseases as either cause or therapeutic target. Regulation of VEGF signaling is therefore of critical importance for many therapeutic strategies. Recent evidence suggests that more VEGF is bound to the extra cellular matrix in vivo than freely diffuses, and that these matrix-binding isoforms of VEGF control branching frequency and network density in vivo. The presentation of VEGF to the receptors by a matrix molecule may initiate a distinct signaling mode, but traditional in vitro cell culture models are unable to differentiate the soluble VEGF signal from the matrix-bound VEGF signal. In this proposal, we develop a combined experimental-computational approach to separate the soluble and matrix-bound VEGF signals. We will develop a novel fibronectin mutant that does not bind VEGF (Aim 1) and we will use state-of-the-art micro patterning techniques to plate endothelial cells on a substrate with gradients of VEGF binding ability (Aim 2). In vitro assays of proliferation, migration and survival will definitively show whether soluble VEGF or matrix-bound VEGF is the more potent stimulator of these critical endothelial cell decisions (Aim 2). We then propose to use in vivo transfection techniques to increase VEGF gradients and to block presentation of matrix-bound VEGF to receptor tyrosine kinases in vivo (Aim 3). This will test whether the in vitro endothelial cell culture is a good model of the decision-making of endothelial cells in vivo. Relevance to Public Health: Inhibition or up regulation of vascular endothelial growth factor (VEGF) is a therapeutic target in over 70 diseases as diverse as cancer, coronary artery disease and diabetic retinopathy. The proposed research uses a novel combined computational and experimental method to elicit how VEGF initiates its signals and, if successful, will better define how to target it therapeutically.

描述(由申请人提供)： 项目摘要：血管内皮生长因子(VEGF)家族由控制血管存活和新生血管萌发的关键细胞因子组成。最近，血管内皮生长因子抑制剂已被FDA批准用于治疗血管增多的疾病(癌症和黄斑变性)，而增加血管内皮生长因子的表达是治疗周围动脉疾病和冠状动脉疾病等缺血性疾病的主要研究途径，尽管临床试验尚未成功。作为病因或治疗靶点，至少有70种疾病与血管内皮生长因子有关。因此，对血管内皮生长因子信号的调节对于许多治疗策略至关重要。最近的证据表明，体内更多的血管内皮生长因子结合在细胞外基质上，而不是自由扩散，这些基质结合的血管内皮生长因子亚型控制着体内的分支频率和网络密度。通过基质分子向受体递呈血管内皮生长因子可能启动一种不同的信号转导方式，但传统的体外细胞培养模型不能区分可溶性的血管内皮生长因子信号和基质结合的血管内皮生长因子信号。在这个建议中，我们开发了一种实验和计算相结合的方法来分离可溶的和基质结合的血管内皮生长因子信号。我们将开发一种不与血管内皮生长因子结合的新型纤维连接蛋白突变体(目标1)，我们将使用最先进的微图案化技术在具有梯度血管内皮生长因子结合能力的底物上电镀内皮细胞(目标2)。体外的增殖、迁移和存活分析将明确表明，在这些关键的内皮细胞决策中，可溶性血管内皮生长因子还是基质结合的血管内皮生长因子是更有效的刺激因子(目标2)。然后，我们建议使用体内转基因技术来增加血管内皮生长因子的梯度，并阻断体内基质结合的血管内皮生长因子对受体酪氨酸激酶的呈递(目标3)。这将检验体外培养内皮细胞是否是体内内皮细胞决策的良好模型。 与公共健康相关：抑制或上调血管内皮生长因子(VEGF)是70多种疾病的治疗目标，包括癌症、冠状动脉疾病和糖尿病视网膜病变。这项拟议的研究使用了一种新的计算和实验相结合的方法来得出血管内皮生长因子是如何启动其信号的，如果成功，将更好地定义如何在治疗上针对它。