MMP-2 in Periodontal Disease and Oral Cancer

申请人描述(申请人提供)：申请人比约恩·斯特芬森，DDS，硕士，博士，正在申请个人科学家奖(K02)，以支持其研究事业发展。这一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症中涉及的酶的方法；2)获得新的蛋白质组学技术方面的新知识和技能，以加强正在进行和未来的研究；3)发展成为一名多产的独立科学家，拥有专注于伤口愈合、牙周病和口腔癌症机制的动态和进步的研究计划。 该研究计划阐述了基质金属蛋白酶-2(MMP2)降解组织的机制。基质金属蛋白酶-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。基质金属蛋白酶是正常发育和组织适应的有益特征，但不受控制的基质金属蛋白酶-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基质金属蛋白酶-2的化合物。由于被基质金属蛋白酶-2切割需要底物分子的结合，我们建议研究基质金属蛋白酶-2结合其主要胶原底物的机制。在与其他科学家的合作中，分子生物学和蛋白质结构分析相结合，首先筛选与基质金属蛋白酶-2的胶原结合结构域(CBD)相互作用的随机多肽文库，并将基质金属蛋白酶-2的结合位点定位在胶原上。随后的核磁共振结构研究将利用含有基质金属蛋白酶-2的CBD和胶原样多肽的复合体来确定基质金属蛋白酶-2上的胶原结合部位。具体的相互作用将通过在已识别的结合位点中引入特定突变来验证。随后，合成肽的生物活性将通过基于结构的修饰而得到增强。 随后的实验将确定合成的生物活性合成肽和结合部位区域是否会抑制基质金属蛋白酶-2并改变细胞在培养中的行为。这些研究将整合新的蛋白质组学方法，以充分了解这些多肽和结合部位区域影响细胞行为的机制。 建议的研究应明确基质金属蛋白酶-2和胶原的特定结合部位，并探索抑制基质金属蛋白酶-2在牙周病和口腔癌中的新策略。

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.