MECHANISMS OF ENDOTHELIAL CELL-COLLAGEN INTERACTIONS

内皮细胞-胶原蛋白相互作用的机制

• 批准号: 7385399
• 负责人: James D SanAntonio
• 金额: $ 4.54万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2008-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7385399
• 关键词: Address; Anabolism; Antibodies; Apical; Binding; Binding Sites; Biological Assay; Blocking Antibodies; Blood Vessels; Blood capillaries; Cell surface; Cells; Chemicals; Chick Embryo; Collagen; Collagen Fibril; Collagen Type I; Data; Development; Developmental Process; Dominant-Negative Mutation; Endothelial Cells; Extracellular Matrix Proteins; Fibrin; Fibronectins; Focal Adhesion Kinase 1; Focal Adhesions; GAG Gene; Gel; Growth Factor; In Vitro; Inorganic Sulfates; Integrin Binding; Integrins; Ischemia; Ligation; Link; MAP Kinase Gene; MAPK14 gene; Malignant Neoplasms; Mediating; Modeling; Morphogenesis; Neoplasm Metastasis; PTK2 gene; Pathology; Pathway interactions; Peptides; Polymers; Proteoglycan; Research; Role; Serum-Free Culture Media; Signal Pathway; Site; Staging; Study models; Support System; Testing; Tube; Unspecified or Sulfate Ion Sulfates; Work; angiogenesis; antibody inhibitor; capillary; chorioallantoic membrane; crosslink; cytokine; extracellular; human disease; in vivo; inhibitor/antagonist; mimetics; monolayer; novel; receptor; response; scaffold; sulfation; tumor growth

Angiogenesis depends upon proper collagen biosynthesis and crosslinking, moreover, type I collagen is an ideal scaffold for angiogenesis in vitro. Despite this, mechanisms of type I collagen-mediated angiogenesis remain poorly understood. We propose to define the features of the type / collagen fibril, endothelial cell surface, and intracellutar signaling pathways that act together to mediate type I collagen- induced angiogenesis. We have developed a unique model for studying endothelial tube morphogenesis in vitro. Our oreliminary data using our system sUPport the hypothe_i_ that engagement between the c_2B1 integrin receotor and integrin-binding sequences of _pe I collagen result in D38MAPK activation, and inactivation of Focal Adhesion Kinase during endothelial tube morphogenesis. We will test this hypothesis in the following aims:l) Define the roles of endothelial cell surface a1_1, a2111, and c_V_3 integrin receptors, sulfated proteoglycans, and fibronectin by testing the activities of integrin or fibronectin function-blocking antibodies and inhibitors of GAG function on angiogenesis; 2) Synthesize triple helical, type I collagen mimetic peptides (THPs) including putative integrin-binding sites, and study their capacities to inhibit cell- collagen attachment, bind integrin receptors, and influence angiogenesis; 3) Study the consequences of integrin function-blocking antibodies, integrin-binding THPs, and chemical and dominant-negative construct inhibitors of signaling pathway function on p38MAPK and FAK activation and angiogenesis; and 4) Examine the role of c_2_1 integrin ligation of discrete collagen sequences, and the p38MAPK and FAK pathways, a) in vivo in the chick CAM, and b) in angiogenesis induction by other polymers including heterotypic collagen fibrils and fibrin. Our work will define the type I collagen structural features critical for its morphogenic activity, probe the functional link between o_2_1 integrin-collagen ligation, p38 MAPK and FAK activation, and langiogenesis, and contribute to the understanding and treatment of human diseases involving vascular insufficiencies, such as ischemia, or abnormal angiogenesis, as in tumor growth.

血管生成依赖于适当的胶原蛋白生物合成和交联，此外，I型胶原蛋白， 胶原是体外血管生成的理想支架。尽管如此，I型胶原介导的机制 血管生成仍然知之甚少。我们建议定义类型/胶原纤维的特征， 内皮细胞表面和细胞内信号通路共同作用，介导I型胶原蛋白- 诱导血管生成。我们已经开发了一个独特的模型，用于研究内皮管形态发生， 体外本系统的初步数据支持c_2B_1和c_2B_2相互作用的假说。 _pe I胶原的整联蛋白受体和整联蛋白结合序列导致D38 MAPK活化， 内皮管形态发生过程中粘着斑激酶失活。我们将测试这个假设， 以下目的：1）确定内皮细胞表面α 1_1、α 2111和c_V_3整联蛋白受体的作用， 硫酸化蛋白聚糖和纤维连接蛋白，通过测试整合素或纤维连接蛋白功能阻断剂的活性， GAG抗体和抑制剂对血管生成的作用; 2）合成三螺旋I型胶原 模拟肽（THP），包括推定的整合素结合位点，并研究其抑制细胞增殖的能力。 胶原附着，结合整合素受体，并影响血管生成; 3）研究 整联蛋白功能阻断抗体、整联蛋白结合THP以及化学和显性阴性构建体 对p38 MAPK和FAK活化和血管生成的信号传导途径功能的抑制剂;和4）检查 c_2_1整合素连接离散胶原序列的作用，以及p38 MAPK和FAK途径，a）在 在鸡CAM中的体内，和B）在通过其它聚合物包括异型胶原的血管生成诱导中 纤维和纤维蛋白。我们的工作将确定I型胶原蛋白的结构特征，这些特征对其形态发生活性至关重要， 探索o_2_1整合素-胶原连接、p38 MAPK和FAK活化之间的功能联系， 血管生成，并有助于了解和治疗人类疾病涉及血管 在某些实施方案中，所述药物组合物可用于治疗诸如局部缺血或异常血管生成（如在肿瘤生长中）的不良反应。