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: 7862651
• 负责人: Feilim C Mac Gabhann
• 金额: $ 24.88万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-10 至 2012-07-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7862651
• 关键词: Adherence; Arteries; Arts; Binding; Blood Vessels; Cell Culture Techniques; Cell Proliferation; Cells; Clinical Trials; Computer Simulation; Coronary Arteriosclerosis; Decision Making; Diabetic Retinopathy; Diffuse; Disease; Endothelial Cells; Environment; Equilibrium; Experimental Designs; Extracellular Matrix; FDA approved; Family; Fibronectins; Frequencies; Genes; In Vitro; Integrins; Left; Ligand Binding Domain; Ligands; Macular degeneration; Malignant Neoplasms; Methods; Modeling; Mus; Muscle Cells; Pathway interactions; Pattern; Peripheral arterial disease; Proliferating; Protein Isoforms; Proteins; Protocols documentation; Public Health; Receptor Activation; Receptor Gene; Receptor Protein-Tyrosine Kinases; Regulation; Relative (related person); Research; Signal Pathway; Signal Transduction; Skeletal Muscle; Techniques; Testing; The Sun; Therapeutic; Tissues; Transfection; Up-Regulation; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factors; base; cell motility; cytokine; density; design; genetic manipulation; in vitro Assay; in vivo; inhibitor/antagonist; migration; mutant; neovascularization; novel; receptor; research study; response; therapeutic target

PROJECT SUMMARY (See Instmctions): 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 extracellular 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 experimentalcomputational approach to separate the soluble and matrix-bound VEGF signals. Using novel or available fibronectin mutants that do not bind VEGF (Aim 1), we will use state-of-the-art micropatteming 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.

项目摘要（参见说明）： 血管内皮生长因子 (VEGF) 家族由参与控制血管存活和血管生成的关键细胞因子组成。最近，VEGF 抑制剂已被 FDA 批准用于治疗血管过多疾病（癌症和黄斑变性），而增加 VEGF 表达是外周动脉疾病和冠状动脉疾病等缺血性疾病的主要研究途径，尽管临床试验正在进行中。 但没有成功。 VEGF 与至少 70 种疾病有关，无论是作为病因还是治疗靶标，因此 VEGF 信号传导的调节对于许多治疗策略至关重要。最近的证据表明，体内与细胞外基质结合的 VEGF 多于自由扩散的 VEGF，并且 VEGF 的这些基质结合亚型控制着体内的分支频率和网络密度。通过基质分子将 VEGF 呈递给受体可能 启动独特的信号传导模式，但传统的体外细胞培养模型无法区分可溶性 来自基质结合 VEGF 信号的 VEGF 信号。在本提案中，我们开发了一种组合实验计算 分离可溶性和基质结合 VEGF 信号的方法。使用不结合 VEGF 的新型或现有纤连蛋白突变体（目标 1），我们将使用最先进的微图案化技术将内皮细胞铺板在具有 VEGF 结合能力梯度的基质上（目标 2）。增殖、迁移和存活的体外测定将明确显示可溶性 VEGF 或基质结合 VEGF 是否是这些关键内皮细胞决策的更有效刺激剂（目标 2）。然后我们建议使用体内转染技术来增加 VEGF 梯度并阻断基质结合的 VEGF 向体内受体酪氨酸激酶的呈递（目标 3）。这将检验体外内皮细胞培养是否是体内内皮细胞决策的良好模型。