Cloaking Key MMP-9 Substrates to Probe the role of Their Cleavage in Plaque Ruptu

描述(申请人提供)：基质金属蛋白酶-9与斑块破裂有关，斑块破裂是心血管疾病临床表现的主要原因。然而，目前还不清楚这种多效性酶裂解哪些关键底物来控制疾病过程。利用再现人类动脉粥样硬化斑块破裂许多特征的独特小鼠模型，我们建议开发潜在的治疗适用的“隐形装置”，以防止在我们已证明被基质金属蛋白酶-9破坏的通路中的两种蛋白质的裂解：凝血级联中的组织因子途径抑制物(TFPI)和参与消退炎症的整合素？2。不是结合和抑制酶，而是专门结合TFPI和整合素2的裂解位点的药物将从已被证明是丰富的蛋白质结合剂来源的类肽的组合文库中鉴定出来。在基于细胞的系统中，将有效地抑制基质金属蛋白酶-9介导的TFPI和整合素？2的蛋白分解，并在必要时增强类肽结合和抑制的亲和力。然后将测试类肽防止斑块破裂的能力，并将有助于确定底物蛋白分解在疾病进展中的生物学功能。探索类肽对底物选择性限制蛋白降解的能力也有可能为心血管疾病提供新的干预策略。 公共卫生相关性：基质金属蛋白酶-9与斑块破裂有关，斑块破裂是心血管疾病的大多数临床表现，但目前尚不清楚这种多效性酶裂解哪些关键底物来控制疾病过程。这项提议的目标是开发和测试潜在的可用于治疗的“隐形装置”，以防止参与凝血和炎症途径的两种关键蛋白质的裂解，我们已经证明，这两种途径被基质金属蛋白酶-9蛋白分解所破坏。这些研究将确定特定底物蛋白降解在疾病进展中的生物学功能，并可能建立一种选择性限制蛋白降解作为心血管疾病潜在干预策略的新方法。

DESCRIPTION (provided by applicant): Matrix metalloproteinase (MMP)-9 is implicated in plaque rupture that is responsible for the majority of clinical manifestations of cardiovascular disease. However, it is still unclear what key substrates this pleiotropic enzyme cleaves to control the disease process. Using a unique mouse model which reproduces many features of human atherosclerotic plaque rupture, we propose to develop potential therapeutically applicable "cloaking devices" to prevent cleavage of two proteins in pathways that we've shown to be disrupted by MMP-9: tissue factor pathway inhibitor (TFPI) in the coagulation cascade and integrin ¿2 involved in the resolution of inflammation. Rather than binding and inhibiting the enzyme, agents that specifically bind the cleavage site of TFPI and integrin ¿2 will be identified from combinatorial libraries of peptide-like peptoids that have been shown to be a rich source of protein-binding agents. Effective inhibition of MMP-9-mediated proteolysis of TFPI and integrin ¿2 will be established in cell-based systems, and the affinity of peptoid binding and inhibition enhanced if necessary. The ability of the peptoids to prevent features of plaque rupture will then be tested, and will help determine the biological function of substrate proteolysis in disease progression. Exploration of the capacity of peptoids to mediate substrate-selective limitation of proteolysis also has the potential to provide new interventional strategies for cardiovascular disease. PUBLIC HEALTH RELEVANCE: Matrix metalloproteinase (MMP)-9 is implicated in plaque rupture that is responsible for the majority of clinical manifestations of cardiovascular disease, but it is still unclear what key substrates this pleiotropic enzyme cleaves to control the disease process. The goal of this proposal is to develop and test potential therapeutically applicable "cloaking devices" to prevent cleavage of two key proteins involved in the coagulation and inflammatory pathways that we've shown to be disrupted by MMP-9 proteolysis. The proposed studies will determine the biological function of specific substrate proteolysis for disease progression, and may establish a new approach for selective limitation of proteolysis as a potential interventional strategy for cardiovascular disease.