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

描述（由申请方提供）：基质金属蛋白酶（MMP）-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的原因。然而，目前还不清楚这种多效性酶切割什么关键底物来控制疾病过程。使用重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”，以防止我们已证明被MMP-9破坏的途径中的两种蛋白质的切割：组织因子途径抑制剂（TFPI）参与凝血级联反应和参与炎症消退的整合素² 2。不是结合和抑制酶，而是从肽样类肽的组合文库中鉴定特异性结合TFPI和整联蛋白2的切割位点的试剂，所述肽样类肽已被证明是蛋白结合剂的丰富来源。将在基于细胞的系统中建立对MMP-9介导的TFPI和整联蛋白2的蛋白水解的有效抑制，并且必要时增强类肽结合和抑制的亲和力。然后将测试类肽预防斑块破裂特征的能力，并将有助于确定底物蛋白水解在疾病进展中的生物学功能。探索类肽介导蛋白水解底物选择性限制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（MMP）-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的原因，但目前仍不清楚这种多效性酶切割什么样的关键底物来控制疾病过程。该提案的目标是开发和测试潜在的治疗适用的“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蛋白质被MMP-9蛋白水解破坏。这些研究将确定特定底物蛋白水解在疾病进展中的生物学功能，并可能建立一种新的方法，选择性限制蛋白水解作为心血管疾病的潜在干预策略。

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.