MMP-2 and Fibronectin Fragmentation in Periodontal Diseases and Oral Cancer

牙周病和口腔癌中的 MMP-2 和纤连蛋白断裂

• 批准号: 7014708
• 负责人: BJORN STEFFENSEN
• 金额: $ 34.68万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7014708
• 关键词: binding sites; fibronectins; mouth neoplasms; peptides; tissues

DESCRIPTION (provided by applicant): The enzymes in the family of matrix metalloproteinases (MMPs) are important to regular tissue maintenance by their collective capacity to degrade major tissue molecules and activating growth factors. The MMPs contribute hereby beneficially to normal development and tissue adaptation. Nevertheless, certain MMPs, including MMP-2, have been found at elevated levels and may abnormally degrade tissues in several diseases, such as periodontal disease, arthritis, and cancer. Recent studies by ourselves and other investigators have shown that the major cell adhesion molecule fibronectin is degraded in those diseases and that MMP-2 can efficiently cleave fibronectin. This degradation requires specific binding between fibronectin and a unique subdomain of MMP-2. Since the behavior of cells changes upon exposure to the fibronectin cleavage fragments, such fragments may alter the disease progression. Therefore, understanding the precise mechanism of binding between the two proteins may provide the basis for developing inhibitors that can block MMP-2 degradation of fibronectin. In a collaborative effort that involves investigators from different disciplines and institutions, we will apply molecular biology, biochemical, mass spectrometry, and protein engineering methods to define the precise binding sites for fibronectin and MMP-2. Based on results gained from screening of phage-displayed random peptide libraries, we will design synthetic peptides, which mimic the binding sites, and test their capacity to block both MMP-2 binding and degradation of fibronectin. Introduction of site-specific mutations in binding domains will confirm the binding sites. After precisely mapping the fibronectin cleavage sites and fragments, we will characterize altered periodontal and cancer cell behavior by multiple parameters, MMP expression, and apoptosis in response to contact with a series of recombinant fibronectin cleavage fragments. We will then test the capacity of the inhibitory peptides to rescue cell functions. This experimental strategy should define the specific binding sites for the interactions between MMP-2 and fibronectin and pursues a novel strategy to inhibit degradation of specific molecules by individual MMPs, such as cleavage of fibronectin by MMP-2. Successful inhibition of fibronectin fragmentation could be of significant benefit for the management of both periodontal disease and oral cancer.

描述（由申请人提供）：基质金属蛋白酶（MMP）家族中的酶通过其降解主要组织分子和激活生长因子的集体能力对于正常组织维护很重要。 MMP 有助于正常发育和组织适应。然而，某些 MMP（包括 MMP-2）被发现水平升高，并且可能在牙周病、关节炎和癌症等多种疾病中异常降解组织。我们和其他研究人员最近的研究表明，主要细胞粘附分子纤连蛋白在这些疾病中被降解，而 MMP-2 可以有效地裂解纤连蛋白。这种降解需要纤连蛋白和 MMP-2 的独特子结构域之间的特异性结合。由于细胞的行为在暴露于纤连蛋白裂解片段后发生变化，因此此类片段可能会改变疾病进展。因此，了解两种蛋白之间结合的精确机制可能为开发能够阻断MMP-2纤连蛋白降解的抑制剂提供基础。在来自不同学科和机构的研究人员的共同努力下，我们将应用分子生物学、生化、质谱和蛋白质工程方法来定义纤连蛋白和 MMP-2 的精确结合位点。根据噬菌体展示的随机肽库筛选结果，我们将设计模拟结合位点的合成肽，并测试其阻断 MMP-2 结合和纤连蛋白降解的能力。在结合域中引入位点特异性突变将确认结合位点。在精确绘制纤连蛋白裂解位点和片段后，我们将通过多个参数、MMP 表达和与一系列重组纤连蛋白裂解片段接触时的细胞凋亡来表征牙周和癌细胞行为的改变。然后我们将测试抑制肽拯救细胞功能的能力。该实验策略应定义 MMP-2 和纤连蛋白之间相互作用的特异性结合位点，并寻求一种新策略来抑制单个 MMP 对特定分子的降解，例如 MMP-2 对纤连蛋白的裂解。成功抑制纤连蛋白断裂可能对牙周病和口腔癌的治疗具有显着的益处。