Biochemistry of MT1-MMP activation in malignancy

恶性肿瘤中 MT1-MMP 激活的生物化学

• 批准号: 8826052
• 负责人: Alex Y Strongin
• 金额: $ 38.84万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8826052
• 关键词: Active Sites; Address; Adult; Agreement; Antibodies; Applied Research; Arabidopsis; Basic Science; Biochemical; Biochemistry; Bioinformatics; Biological Models; Biology; Biosensor; Cancer Biology; Cell membrane; Cell model; Cell physiology; Cell surface; Cell-Cell Adhesion; Cells; Cleaved cell; Clinical; Computer Simulation; Consensus; Data; Detection; Development; Diagnostic Neoplasm Staging; Disease; Drug Targeting; Dwarfism; Enzyme Precursors; Enzymes; Event; Exhibits; Extracellular Matrix Proteins; Funding; Gelatinase A; Genes; Grant; Human; Immunodeficient Mouse; In Vitro; Knockout Mice; Knowledge; Lead; Length; Link; Location; MMP14 gene; Malignant - descriptor; Malignant Neoplasms; Matrix Metalloproteinases; Membrane Proteins; Molecular Models; Mus; Neoplasm Metastasis; Normal Cell; Oncogenes; Pathology; Pathway interactions; Peer Review; Peptide Hydrolases; Phenotype; Plants; Play; Process; Proteolysis; Publications; Publishing; Recycling; Regulation; Research; Role; Scientist; Signal Transduction; System; Testing; Tumor Cell Biology; Tumor Cell Invasion; Tumor stage; Tumorigenicity; United States National Institutes of Health; Work; base; biochemical model; cancer cell; cell motility; cell type; design; human MMP14 protein; in vivo; inhibitor/antagonist; member; membrane-type matrix metalloproteinase; molecular modeling; neoplastic cell; novel; outcome forecast; prevent; programs; proteinase In; receptor; success; trafficking; tumor; tumor growth; tumor progression; tumor xenograft; tumorigenic

DESCRIPTION (provided by applicant): This application is a based on our previously funded 5-year project that generated over 40 peer-reviewed publications. Invasion-promoting, pro-tumorigenic MT1-MMP is implicated as the most important MMP in cancer biology and is known to play specialized roles as a modifier of cell function. There is a consensus among scientists that MT1-MMP is a key player in cell surface proteolytic events. We now know that in multiple model systems MT1-MMP acts as an oncogene, stimulates tumor cell invasion and metastasis, and takes over tumor growth control. In agreement, in humans MT1-MMP activity correlates significantly with metastases, clinical stage, and tumor size. Our knowledge of the MT1-MMP biology, however, is limited as yet especially when compared with the role this proteinase plays in malignancy. As of now, the mechanisms of MT1-MMP activation are not completely understood. As a result of our study we can now suggest that rather than exhibiting a uniform activation pathway in any cell type, invasion-promoting MT1-MMP appears to have an activation mechanism hierarchy. To increase their proteolytic arsenal, tumor cells exploit the mechanism leading to the complete inactivation of the MT1-MMP's autoinhibitory prodomain while in normal cells MT1- MMP activity is controlled by the intact prodomain that is released by furin cleavage alone. As a result, the intact prodomain prevents the potentially damaging function of pro-invasive, pro-tumorigenic MT1-MMP in the normal microenvironment. Autocatalytically activated furin plays an important role in the processing and activation of multiple proproteins including MT1-MMP and MMP-2, the downstream target of MT1-MMP. Because of its importance in cancer, MT1-MMP is a promising drug target in disease and its selective biosensor and inhibitors are urgently needed. Overall, it is obvious that additional work is needed to gain a better understanding of the MT1-MMP structural-functional relationships. Our three Specific Aims, which we have designed to achieve a better understanding of MT1-MMP are: (1) Determine the functional significance of a two-step prodomain processing mechanism of MT1-MMP activation in normal and cancer cells, (2) Design and test efficient and selective, cleavage-activated biosensors of MT1-MMP and furin (an activator of MT1-MMP), and (3) Design and evaluate the anti-tumor activity of the highly selective and potent, active site- targeting, inhibitory Fab/scFv human antibodies to MT1-MMP. The result of our proposed studies will be a thorough understanding of the many roles played by MT1-MMP and its activator, furin. Our program will use an integrated systems approach involving biochemical, molecular, cellular and computer modeling studies. We will also conduct studies using tumor xenografts in mice to validate our results gained in vitro. Our program will result in a refined understanding of how malignant cells traffic from their original location and spread throughout the body. This valuable information will lead directly to the development of novel and effective strategies for the detection, prognosis, and treatment of a broad range of malignancies.

描述（由申请人提供）：本申请是基于我们以前资助的5年项目，该项目产生了40多篇同行评审的出版物。 促侵袭、促肿瘤发生的MT 1-MMP被认为是癌症生物学中最重要的MMP，并且已知其作为细胞功能的修饰剂发挥专门作用。 科学家们一致认为MT 1-MMP是细胞表面蛋白水解事件的关键参与者。 我们现在知道，在多个模型系统中，MT 1-MMP作为癌基因，刺激肿瘤细胞侵袭和转移，并接管肿瘤生长控制。 一致的是，在人类中，MT 1-MMP活性与转移、临床分期和肿瘤大小显着相关。 然而，我们对MT 1-MMP生物学的了解是有限的，特别是与这种蛋白酶在恶性肿瘤中所起的作用相比。 到目前为止，MT 1-MMP激活的机制还不完全清楚。 作为我们研究的结果，我们现在可以表明，而不是在任何细胞类型中表现出统一的激活途径，促进侵袭的MT 1-MMP似乎有一个激活机制层次。 为了增加它们的蛋白水解武库，肿瘤细胞利用导致MT 1-MMP的自身抑制性前结构域完全失活的机制，而在正常细胞中，MT 1- MMP活性由单独通过弗林蛋白酶切割释放的完整前结构域控制。 因此，完整的前结构域防止了正常微环境中促侵袭、促肿瘤发生MT 1-MMP的潜在破坏性功能。 自催化活化的弗林蛋白酶在多种前体蛋白的加工和活化中起重要作用，包括MT 1-MMP和MMP-2，其是MT 1-MMP的下游靶标。 由于其在肿瘤中的重要性，MT 1-MMP是一个很有前途的药物靶点，其选择性生物传感器和抑制剂的需求是迫切的。 总体而言，很明显需要进行额外的工作才能更好地了解MT 1-MMP的结构-功能关系。 我们的三个具体目标，我们已经设计，以实现更好地了解MT 1-MMP是：（1）确定正常和癌细胞中MT 1-MMP激活的两步前结构域加工机制的功能意义，（2）设计和测试有效和选择性的MT 1-MMP和弗林蛋白酶的切割激活的生物传感器（MT 1-MMP的激活剂）的抗肿瘤活性的研究;（3）设计并评价抗MT 1-MMP的高选择性和有效的、活性位点靶向的、抑制性Fab/scFv人抗体的抗肿瘤活性。 我们提出的研究的结果将是一个由MT 1-MMP和它的激活剂，弗林蛋白酶发挥的许多作用的透彻理解。 我们的计划将使用一个综合系统的方法，涉及生物化学，分子，细胞和计算机建模研究。 我们还将使用小鼠肿瘤异种移植物进行研究，以验证我们在体外获得的结果。 我们的计划将导致对恶性细胞如何从其原始位置运输并在整个身体中扩散的精确理解。 这些有价值的信息将直接导致开发新的和有效的策略，用于检测，预后和治疗广泛的恶性肿瘤。