MMP-2 in Periodontal Disease and Oral Cancer

描述（由申请人提供）：申请人 Bjorn Steffensen，DDS，MS，PhD，正在申请个人科学家奖（K02）以支持他的研究职业发展。该职业发展计划的目标是：1) 扩展正在进行的研究，以确定抑制牙周病和口腔癌相关酶的方法，2) 获得新型蛋白质组学技术的新知识和技能，以加强正在进行和未来的研究，3) 发展成为一名富有成效的独立科学家，开展动态和渐进的研究计划，重点关注伤口愈合、牙周病和口腔癌的机制。 该研究计划探讨基质金属蛋白酶-2 (MMP-2) 降解组织的机制。 MMP-2 属于基质金属蛋白酶家族，它们共同可以降解大多数组织成分。 MMP 是正常发育和组织适应的有益特征，但不受控制的 MMP-2 活性与炎症性疾病、癌症和伤口愈合不良密切相关。 该提案旨在开发能够特异性抑制MMP-2的化合物。由于 MMP-2 的裂解需要结合底物分子，因此我们建议研究 MMP-2 结合其主要胶原底物的机制。与其他科学家合作，结合分子生物学和蛋白质结构分析，首先筛选与 MMP-2 的胶原蛋白结合域 (CBD) 相互作用的随机肽库，并绘制胶原蛋白上的 MMP-2 结合位点图谱。随后的核磁共振结构研究将使用含有来自 MMP-2 的 CBD 和类胶原肽的复合物来识别 MMP-2 上的胶原蛋白结合位点。通过将特定突变引入已识别的结合位点来验证特定的相互作用。随后，合成肽的生物活性将通过基于结构的修饰得到增强。 后续实验将确定合成的生物活性合成肽和结合位点区域是否可以抑制 MMP-2 并改变培养物中细胞的行为。这些研究将整合新颖的蛋白质组学方法，以充分了解此类肽和结合位点区域影响细胞行为的机制。 拟议的研究应明确MMP-2和胶原蛋白的特异性结合位点，并探索在牙周病和口腔癌中抑制MMP-2的新策略。

DESCRIPTION (provided by applicant): The applicant, Bjorn Steffensen, DDS, MS, PhD, is applying for the Individual Scientist Award (K02) to support his research career development. The goals of this career development program are to: 1) Extend the ongoing research to identify approaches to inhibit enzymes involved in periodontal disease and oral cancer, 2) Gain new knowledge and skills on novel proteomics technologies to strengthen ongoing and future research, and 3) Evolve into a productive and independent scientist with a dynamic and progressive research program focused on the mechanisms of wound healing, periodontal disease, and oral cancer. The research plan addresses the mechanism by which matrix metalloproteinase-2 (MMP-2) degrades tissues. The MMP-2 belongs to the matrix metalloproteinase family of enzymes, which collectively can degrade most tissue components. The MMPs are beneficial features of normal development and tissue adaptation, but uncontrolled MMP-2 activity has been associated strongly with inflammatory diseases, cancer, and poor wound healing. The proposal is designed to develop compounds that can specifically inhibit MMP-2. Since cleavage by the MMP-2 requires binding of the substrate molecules, we propose to investigate the mechanism by which MMP-2 binds its main collagen substrate. In collaboration with other scientists, molecular biology and protein structural analyses are combined to first screen random peptide libraries for interaction with the collagen-binding domain (CBD) of MMP-2 and map the MMP-2 binding sites on collagen. Subsequent nuclear magnetic resonance structural studies will identify the collagen binding site on the MMP-2 using complexes containing the CBD from MMP-2 and collagen-like peptides. The specific interactions will be verified by introducing specific mutations into the identified binding site. Subsequently the bioactivity of the synthetic peptides will be enhanced by structure-based modifications. Subsequent experiments will determine whether the synthetic bioactive synthetic peptides and binding site regions may inhibit MMP-2 and alter the behavior of cells in cultures. The studies will integrate novel proteomics approaches to fully understand the mechanisms by which such peptides and binding site regions influence cell behavior. The proposed studies should define the specific binding sites of MMP-2 and collagen and explore a new strategy to inhibit MMP-2 in periodontal disease and oral cancer.