MECHANISMS OF ENDOTHELIAL CELL-COLLAGEN INTERACTIONS

内皮细胞-胶原蛋白相互作用的机制

• 批准号: 6739011
• 负责人: James D SanAntonio
• 金额: $ 31.4万
• 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 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6739011
• 关键词: angiogenesis; antibody; biological signal transduction; cell membrane; chick embryo; chorioallantoic membrane; collagen; enzyme activity; fibrin; fibronectins; focal adhesion kinase; integrins; intermolecular interaction; laboratory rat; mitogen activated protein kinase; mucopolysaccharides; protein structure function; proteoglycan; tissue /cell culture

DESCRIPTION (provided by applicant): 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 intracellular 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 preliminary data using our system support the hypothesis that engagement between the alpha2beta1 integrin receptor and integrin-binding sequences of type 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:1) Define the roles of endothelial cell surface alpha1beta1, alpha2beta1, and alphaVbeta3 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 alpha2beta1 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 alpha2beta1 integrin-collagen ligation, p38 MAPK and FAK activation, and angiogenesis, and contribute to the understanding and treatment of human diseases involving vascular insufficiencies, such as ischemia, or abnormal angiogenesis, as in tumor growth.

描述（申请人提供）：血管生成取决于适当的胶原蛋白生物合成和交联，而且，I型胶原蛋白是体外血管生成的理想支架。尽管如此，I 型胶原介导的血管生成机制仍然知之甚少。我们建议定义 I 型胶原原纤维、内皮细胞表面和细胞内信号通路的特征，这些通路共同作用以介导 I 型胶原诱导的血管生成。我们开发了一种独特的模型来研究体外内皮管形态发生。我们使用我们的系统的初步数据支持以下假设：在内皮管形态发生过程中，α2β1 整合素受体和 I 型胶原整合素结合序列之间的结合导致 D38MAPK 激活和粘着斑激酶失活。我们将通过以下目的检验这一假设：1）通过测试整合素或纤连蛋白功能阻断抗体和GAG功能抑制剂对血管生成的活性，定义内皮细胞表面α1β1、α2β1和αVβ3整合素受体、硫酸化蛋白聚糖和纤连蛋白的作用； 2) 合成三螺旋、I 型胶原模拟肽（THP），包括假定的整合素结合位点，并研究其抑制细胞胶原附着、结合整合素受体和影响血管生成的能力； 3) 研究整合素功能阻断抗体、整合素结合THP以及信号通路功能的化学和显性失活结构抑制剂对p38MAPK和FAK激活和血管生成的影响； 4) 检查离散胶原序列的 α2β1 整合素连接以及 p38MAPK 和 FAK 途径的作用，a) 在小鸡 CAM 体内，b) 在其他聚合物（包括异型胶原原纤维和纤维蛋白）诱导血管生成中的作用。我们的工作将定义对其形态发生活性至关重要的 I 型胶原结构特征，探讨 α2β1 整联蛋白-胶原连接、p38 MAPK 和 FAK 激活以及血管生成之间的功能联系，并有助于理解和治疗涉及血管功能不全的人类疾病，例如缺血或肿瘤生长中的异常血管生成。