Regulation of MT-MMPs by Trafficking in Cancer Cells

癌细胞贩运对 MT-MMP 的调节

基本信息

批准号：
7163439
负责人：
DUANQING PEI
金额：
$ 22.54万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-12-05 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7163439
关键词：
3-Dimensional Animal Model Back Benign Biological Models Cell Surface Receptors Cell membrane Cell surface Cells Chemoprevention Chimera organism Clathrin-Coated Vesicles Collagen Type I Cytoplasmic Tail Development Doctor of Philosophy Down-Regulation Dynamin Early Endosome Endocytosis Endopeptidases Enzymes Epigenetic Process Excision Extracellular Matrix Genetic Golgi Apparatus Growth Growth Factor Invaded Invasive Lead Lysosomes MT3 gene Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Matrix Metalloproteinases Measures Mediating Neoplasm Metastasis Peptide Hydrolases Pharmacotherapy Phenotype Play Process Recycling Regulation Research Personnel Role Series Specimen TFAP2A gene Testing Time Transmembrane Domain Tumor Cell Invasion cancer cell cell growth extracellular gain of function human MMP14 protein insight late endosome member membrane-type matrix metalloproteinase mutant neoplastic cell novel proMMP-2 programs trafficking

项目摘要

Our long term objective is to determine how membrane-type matrix metalloproteinases (MT-MMPs) regulate tumor invasion and metastasis. Although matrix metalloproteinases (MMPs) have been implicated in the progression of malignant cancer, the mechanism by which tumor cells deploy these destructive proteinases remains unresolved. MT-MMPs are a subset of MMPs with transmembrane domains capable of focusing these enzymes on plasma membrane against extracellular matrix (ECM) barriers, representing a novel paradigm on how tumor cells deploy proteolytic activity during tumor invasion and metastasis. We have recently demonstrated that MT1-MMP is regulated by endocytosis in clathrin- coated vesicles to control its proteolytic activity on cell surface. We established that its cytoplasmic domain is required for endocytosis and its removal gave rise to a gain-of-function mutant of this enzyme. This endocytosis-deficient mutant is more potent than wild type MT1-MMP in promoting invasiveness in prostate cancer cells, suggesting that the invasiveness of tumor cells can be regulated through the trafficking of MT-MMPs. Employing a chimera between MT1-MMP and MT3-MMP, we obtained the first evidence that internalized MT-MMPs can be recycled back to cell surface. In this application, we propose to integrate the studies of MT1-MMP and MT3- MMP to test the hypothesis that tumor cells modulate their invasiveness by controlling the trafficking of MT- MMPs through their cytoplasmic domains. First, we will define the trafficking of MT1-MMP and MT3-MMP from cell surface to various intracellular compartmetns and characterize the impact of growth factors and extracellular matrix on their trafficking. Secondly, we will define the role of cytoplasmic domains of MT1- MMP and MT3-MMP in mediating their trafficking/recycling and characterize their interactions with cellular components. Thirdly, we will define the regulatory role of trafficking on MT1-MMP and MT3-MMP mediated cell growth and invasion within or through 3-dimensional type I collagen matrix. Results from these approaches may yield insights on how tumor cells gain a growth and invasive advantage by regulating the trafficking of MT-MMPs and such insights may lead to the development of novel drugs for therapy and chemoprevention against cancer.

我们的长期目标是确定膜型基质金属蛋白酶 (MT-MMP) 如何调节肿瘤的侵袭和转移。尽管基质金属蛋白酶 (MMP) 与恶性癌症的进展，肿瘤细胞部署这些破坏性蛋白酶的机制仍未解决。 MT-MMP 是 MMP 的一个子集，具有能够聚焦的跨膜域这些位于质膜上的酶对抗细胞外基质（ECM）屏障，代表了一种新颖的肿瘤细胞在肿瘤侵袭和转移过程中如何发挥蛋白水解活性的范例。我们有最近证明，MT1-MMP 通过网格蛋白包被的囊泡中的内吞作用来调节其细胞表面的蛋白水解活性。我们确定其胞质结构域是内吞作用所必需的其去除产生了该酶的功能获得突变体。这种内吞作用缺陷的突变体是比野生型 MT1-MMP 更有效地促进前列腺癌细胞的侵袭，这表明肿瘤细胞的侵袭性可以通过 MT-MMP 的运输来调节。使用嵌合体在 MT1-MMP 和 MT3-MMP 之间，我们获得了第一个证据，证明内化的 MT-MMP 可以回收回到细胞表面。在此应用中，我们建议整合 MT1-MMP 和 MT3- MMP 检验肿瘤细胞通过控制 MT- 的运输来调节其侵袭性的假设 MMP 通过其细胞质结构域。首先，我们来定义MT1-MMP和MT3-MMP的贩运从细胞表面到各个细胞内区室，并表征生长因子和细胞外基质对其贩运的影响。其次，我们将定义 MT1-细胞质结构域的作用 MMP 和 MT3-MMP 介导其运输/回收并表征其与细胞的相互作用成分。第三，我们将定义贩运对MT1-MMP和MT3-MMP介导的调节作用细胞在 3 维 I 型胶原基质内或通过 3 维 I 型胶原基质生长和侵袭。这些结果这些方法可能会深入了解肿瘤细胞如何通过调节 MT-MMP 的贩运和此类见解可能会导致用于治疗和治疗的新药物的开发化学预防癌症。