Spatial Control of Matrix Proteolysis in Breast Cancer

Institution: The Burnham Institute for Medical Research
Investigator(s): Alex Strongin, Ph.D. - Alex Strongin, Ph.D. - Alex Strongin, Ph.D. -
Award Cycle: 1999 (Cycle V) Grant #: 5JB-0094A Award: $340,383
Award Type: IDEAS II
Research Priorities
Biology of the Breast Cell>Pathogenesis: understanding the disease

This is a collaboration with: 5JB-0094 - 5JB-0094 -

Initial Award Abstract (1999)
The long-term objectives of our proposed research are to enhance understanding of the pathogenesis of breast cancer and to develop more effective interventions for treating this disease. There are strong correlations between breast cancer progression, invasion, metastasis and metalloproteinase activity in breast tumors. Metalloproteinases (MMPs) are the enzymes essential for the degradation of the matrix surrounding tumor cells that enable tumors to metastasize and grow in size. To control breast cancer, it appears necessary to regulate the MMPs expressed in most invasive breast tumors. In addition, tumors carry specific receptors capable of promoting the adhesion and invasion of breast cancer cells. The focus of the present study is to explore mechanisms by which MMPs can be localized to the cell surface of breast cancer cells to maximize their activity in facilitating cell migration. Our hypothesis is that the integrin receptor avb3 is the prime candidate for interacting with MMPs, called MT1-MMP and MMP-2.

Thus, our aims are designed to (1) understand how MMPs modify receptors of breast cancer cells, (2) identify the mechanisms and components capable of delivering MMPs to the discrete regions at the cell surface in an immediate proximity to these cell receptors, and (3) determine how we can employ this knowledge to design efficient inhibitors of tumor cell locomotion. These experiments will be performed by co-expressing MT-1-MMP and the integrin b3 subunit proteins in breast carcinoma cell lines. Using this approach we will be able to determine whether the integrin adhesion receptor and the MMP interact with each other and whether there are localized proteolysis effects leading to enhanced cellular movement.

These models will provide us with a challenging opportunity to discriminate and specify the effects of individual components involved in the locomotion of tumor cells. Understanding these mechanisms in should ultimately provide clues to facilitate the design of novel efficient inhibitors of focal proteolysis and breast cancer cell invasion

Final Report (2001)
The main objectives of our project are to functionally and structurally characterize a cross-talk between matrix metalloproteinases (MMPs), cell surface receptors and cytoskeleton that occurs in migrating breast cancer cells and to suggest inhibitors capable of favorably modifying individual components involved in these complex chain of interactions. Specifically, our aims were to evaluate modifications of cell receptors such as integrins by membrane type-1 MMP (MT1-MMP) and to identify mechanisms that are capable of localizing MMPs through their direct binding associations with certain intracellular component(s) to discrete regions of cell surface. Ultimately, these mechanisms may be the targets for novel pharmaceuticals capable of specifically regulating the geography of the focalized proteolysis in breast cancer.

We successfully accomplished our aims. Specifically, we identified mechanisms involved in the regulation and activation of metalloproteinases such as MT1-MMP and metalloproteinase-2 (MMP-2) in breast carcinoma cells. Further, we characterized the processing of the precursor of integrin aV chain (pro-aV) by MT1-MMP and presented evidence of an alternative, furin-independent pathway of pro-a v processing by this protease. Processing of pro-a v by MT1-MMP significantly facilitates integrin-mediated adhesion and migration of cells. The resulting integrin more efficiently mediates outside-in signal transduction through a focal adhesion kinase (FAK) pathway relative to the conventional integrin. These novel regulatory mechanisms may exist in the most aggressive carcinoma tumor types. These mechanisms involved in the processing of integrin a subunits underscore the significance and complexity of interactions between MT1-MMP and adhesion receptors, and suggest for the first time that regulation of integrin functionality may be an important role of MT1-MMP in migrating tumor cells. Further, our findings suggest a mechanism of selective docking of MMP-2 via the enzyme’s C-terminal hemopexin-like domain at tumor cell surfaces, specifically at the sites that include MT1-MMP and activated integrin a vb 3.

In addition, we identified the multifunctional, ubiquitously expressed chaperon-like protein, gC1qR/p33, as a protein that interacts with the cytoplasmic tail domain of MT1-MMP. The data from immunoprecipitation, phase fractionation, immunofluorescence and inhibitory studies suggest that gC1qR/p33 regulates trafficking of MT1-MMP in breast carcinoma cells. Binding with gC1qR/p33 tethers MT1-MMP intracellularly, while its cleavage appeares to release the proteinase. The MT1-MMP· gC1qR/p33 interactions are likely to have a pleiotropic effect on cell physiology by modulating both the compartmentalization and trafficking of MT1-MMP. These mechanisms may provide a link between spatial regulation of focal proteolysis by the cell surface associated MMPs and the regulation of integrin-mediated motility of tumor cells. Our experimental results, which were summarized in several publications in peer-reviewed journals may have a direct impact on a design of novel inhibitors and facilitate clinical trials involving the existing synthetic inhibitors of MMPs.

Functional activation of integrin avb3 in tumor cells expressing membrane-type 1 matrix metalloproteinase
Periodical:International Journal of Cancer
Index Medicus: Int J Cancer
Authors: Deryugina EI, Bourdon MA, Jungwirth K, Smith JW, and Strongin AY
Yr: 2000 Vol: 86 Nbr: Abs: Pg:15-23

MT1-MMP Initiates Activation of pro-MMP-2 and Integrin avb3 Promotes Maturation of MMP-2 in Breast Carcinoma Cells
Periodical:Experimental Cell Research
Index Medicus: Exp Cell Res
Authors: Dreyugina EI, Ratnikov B, Monosov E, Postnova TI, DiScipio R, Smith JW, and Strongin AY
Yr: 2001 Vol: 263 Nbr: Abs: Pg:209-223

Characterization of Matrix Metalloproteinase-26, a Novel Metalloproteinase Widely Expressed in Cancer Cells of Epithelial Origin
Periodical:Biochemical Journal
Index Medicus: Biochem J
Authors: Marchenko GN, Ratnikov BI, Rozanov DV, Godzik A, Deryugina EI, and Strongin AY
Yr: 2001 Vol: 356 Nbr: Abs: Pg:705-718

MMP-28, a new Human Matrix Metalloproteinase with an Unusual Cysteine-switch Sequence in Widely Expressed in Tumors
Index Medicus: Gene
Authors: Marchenko GN and Strongin AY
Yr: 2001 Vol: 265 Nbr: Abs: Pg:87-93

Matrix-dependent Proteolysis of Surface Transglutaminase by Membreane-type Metalloproteinase Regulates Cancer Cell Adhesion and Locomotion
Periodical:Journal of Biological Chemistry
Index Medicus: J Biol Chem
Authors: Belkin AM, Ajimov SS, Zaritskaya LS, Ratnikov BI, Deryugina EI, and Strongin AY
Yr: 2001 Vol: 276 Nbr: Abs: Pg:18415-18422

Determination of Matrix Metalloproteinase Activity Using Biotinylated Gelatin
Periodical:Analytical Biochemistry
Index Medicus: Anal Biochem
Authors: Ratnikov B, Deryugina E, Leng J, Marchenko G, Dembrow D, anf Strongin A
Yr: 2000 Vol: 286 Nbr: Abs: Pg:149-155

Mutation Analysis of Membreane Type-1 matrix Metalloproteinase (MT-1-MMP)
Periodical:Journal of Biological Chemistry
Index Medicus: J Biol Chem
Authors: Rozanov DV, Deryugina EI, Ratnikov BI, Monsov EZ, Marchenko GN, Quigley JP, Strongin AY
Yr: 2001 Vol: 276 Nbr: Abs: Pg:25705-25714