Biochemistry of MT1-MMP activation in malignancy

恶性肿瘤中 MT1-MMP 激活的生物化学

基本信息

批准号：
8270459
负责人：
Alex Y Strongin
金额：
$ 38.05万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
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/8270459
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 Delivery Systems 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

项目摘要

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）：该申请基于我们之前资助的 5 年项目，该项目产生了 40 多篇同行评审出版物。促进侵袭、促肿瘤发生的 MT1-MMP 被认为是癌症生物学中最重要的 MMP，并且已知作为细胞功能调节剂发挥着特殊作用。科学家们一致认为 MT1-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，MT1-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。一致认为，在人类中，MT1-MMP 活性与转移、临床分期和肿瘤大小显着相关。然而，我们对 MT1-MMP 生物学的了解还很有限，特别是与这种蛋白酶在恶性肿瘤中发挥的作用相比。迄今为止，MT1-MMP 激活的机制尚不完全清楚。根据我们的研究结果，我们现在可以表明，促进侵袭的 MT1-MMP 似乎具有激活机制层次结构，而不是在任何细胞类型中表现出统一的激活途径。为了增加其蛋白水解库，肿瘤细胞利用导致 MT1-MMP 的自抑制前结构域完全失活的机制，而在正常细胞中，MT1-MMP 活性由仅由弗林蛋白酶裂解释放的完整前结构域控制。因此，完整的前结构域可防止正常微环境中促侵袭、促肿瘤发生的 MT1-MMP 的潜在破坏性功能。自催化激活的弗林蛋白酶在多种前蛋白的加工和激活中发挥着重要作用，包括 MT1-MMP 和 MMP-2（MT1-MMP 的下游靶标）。由于其在癌症中的重要性，MT1-MMP 是一种有前途的疾病药物靶点，迫切需要其选择性生物传感器和抑制剂。总的来说，很明显需要做更多的工作来更好地理解 MT1-MMP 结构功能关系。我们为更好地理解 MT1-MMP 而设计的三个具体目标是：(1) 确定正常细胞和癌细胞中 MT1-MMP 激活的两步前结构域处理机制的功能意义，(2) 设计并测试高效、选择性、裂解激活的 MT1-MMP 和弗林蛋白酶（MT1-MMP 激活剂）生物传感器，以及 (3) 设计和评估高度选择性和裂解激活生物传感器的抗肿瘤活性。 MT1-MMP 的有效、活性位点靶向、抑制性 Fab/scFv 人类抗体。我们提出的研究结果将是对 MT1-MMP 及其激活剂弗林蛋白酶所发挥的多种作用的全面了解。我们的项目将使用涉及生化、分子、细胞和计算机建模研究的集成系统方法。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。我们的项目将深入了解恶性细胞如何从其原始位置转移并扩散到全身。这些有价值的信息将直接导致开发新颖有效的策略来检测、预后和治疗多种恶性肿瘤。