Targeting the PEX Domain of MT1-MMP: Novel Cancer Therapy

上皮向间充质转化(EMT)已成为癌症早期的关键步骤。更好 对EMT机制的了解将促进抑制物预防癌症的发展 进步。大量证据表明，膜型1-基质金属蛋白酶(MT1-MMPs)与血管内皮细胞瘤 EMT和转化为侵袭性癌症。MT1-MMPproMMP2的激活及其降解 细胞外基质(ECM)成分参与了癌症的进展。MT1-MMPs还切割了许多 细胞表面蛋白、生长因子和趋化因子。我们最近证明了MT1-MMP1：1)裂解 2)促进细胞在3DI型胶原凝胶中的迁移/分散； 3)在体外和体内调节细胞增殖；5)改变3DI型胶原凝胶的细胞形态。我们 同时也证明了MT1-MMPPEX结构域在癌细胞迁移中起着关键作用 并精确定位了MT1-MMPs介导的细胞迁移所需的区域。以PEX领域为目标 重组MT1-MMPPEX蛋白干扰MT1-MMPPEX诱导的细胞迁移。这个 这项资助的主要目的是确定MT1-基质金属蛋白酶的催化和非催化活性在 急诊室。这一特征将有助于我们开发特定的非催化结构域抑制剂 MT1-基质金属蛋白酶将被用来干扰癌症的进展。为了实现这一目标， 将检测3D培养的癌细胞EMT中MT1-MMPs的表达。EMT相关转录程序和 将对信号通路进行评估。基于MT1-MMPs的计算模型及其与其他组织的相似性 将确定癌细胞迁移所需的MMPs，MT1-MMPPEX结构域中的最小基序(S) 使用一种诱变方法。通过PEX形成MT1-基质金属蛋白酶的均二聚体和/或杂寡体 域名将被确定。基于关键PEX基序的鉴定，特定的MT1-基质金属蛋白酶抑制 多肽将使用分析化学和细胞生物学方法进行设计和表征。 功能抑制肽将在体内癌症模型中生产和评估。长期目标是 开发一种先导化合物，该化合物将被修饰以产生一种副作用最小的MT1-MMPI抑制药物 治疗癌症的效果。我认为目前的项目将帮助我们更好地了解癌症 这项研究取得了进展，并导致了治疗早期癌症的新的基本工具的开发。

Epithelial-to-mesenchymal transition (EMT) has emerged as a critical step in the early stage of cancer. Better understanding of the mechanism of EMT will foster the development of inhibitors to prevent cancer progression. Considerable evidence has implicated membrane type 1-matrix metalloproteinase (MT1-MMP) in EMT and conversion to aggressive cancers. MT1-MMP activation of proMMP-2 and degradation of extracellular matrix (ECM) components are involved in cancer progression. MT1-MMP also cleaves numerous cell surface proteins, growth factors, and chemokines. We recently demonstrated that MT1-MMP: 1) cleaves E-cadherin at the cell-cell adherins junctions; 2) promotes cell migration/scattering in 3D type I collagen gels; 3) regulates cell proliferation in vitro and in vivo; and 5) changes cell morphology in 3D type I collagen gels. We also demonstrated that the hemopexin (PEX) domain of MT1-MMP plays a crucial role in cancer cell migration and pinpointed the regions required for MT1-MMP-mediated cell migration. Targeting the PEX domain with recombinant MT1-MMP PEX protein resulted in interference with MT1-MMP-induced cell migration. The primary goal of this grant is to define the involvement of the catalytic and non catalytic activities of MT1 -MMP in EMT. This characterizationwill facilitate our development of specific non-catalytic domain inhibitors of MT1-MMP that will be employed to interfere with cancer progression. To achieve this aim, the function of MT1-MMP in EMT in 3D cultured cancer cells will be examined. EMT-related transcription programs and signaling pathways will be evaluated. Based on a computational model of MT1-MMP and similarity with other MMPs, minimum motif(s) in the PEX domain of MT1-MMP required for cancer cell migration will be identified using a mutagenesis approach. Formation of homodimer and/or heterooligomer of MT1-MMP through PEX domain will be determined. Based on the identification of crucial PEX motifs, specific MT1-MMP inhibitory peptides will be designed and characterized using analytical chemistry and cell biological approaches. Functional inhibitory peptides will be produced and evaluated in an in vivo cancer model. The long-term goal is to develop a lead compound that will be modified to produce a MT1-MMP inhibitory drug with minimal side effects for treatment of cancer. I propose that the current project will help us to better understand cancer progression and lead to the development of novel basic tools for treatment of early stage cancer.