Discoidin domain receptor 2 mediates collagen-induced activation of membrane-type 1 matrix metalloproteinase in human fibroblasts

Iwona Majkowska; Yasuyuki Shitomi; Noriko Ito; Nathanael S. Gray; Yoshifumi Itoh

doi:10.1074/jbc.M116.770057

Discoidin domain receptor 2 mediates collagen-induced activation of membrane-type 1 matrix metalloproteinase in human fibroblasts

From the ^‡Kennedy Institute of Rheumatology, Kennedy Institute of Rheumatology, University of Oxford, Roosevelt Drive, Oxford OX3 7FY, United Kingdom and
^§Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02215

↵1 To whom correspondence should be addressed: Kennedy Institute of Rheumatology, Nuffield Dept. of Orthopaedics, Rheumatology, and Musculoskeletal Sciences, University of Oxford, Roosevelt Dr., Headington, Oxford OX3 7FY, UK. E-mail: yoshi.itoh{at}kennedy.ox.ac.uk.

Edited by Amanda J. Fosang

Abstract

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a membrane-bound MMP that is highly expressed in cells with invading capacity, including fibroblasts and invasive cancer cells. However, pathways of MT1-MMP up-regulation are not clearly understood. A potential physiological stimulus for MT1-MMP expression is fibrillar collagen, and it has been shown that it up-regulates both MT1-MMP gene and functions in various cell types. However, the mechanisms of collagen-mediated MT1-MMP activation and its physiological relevance are not known. In this study, we identified discoidin domain receptor 2 (DDR2) as a crucial receptor that mediates this process in human fibroblasts. Knocking down DDR2, but not the β1 integrin subunit, a common subunit for all collagen-binding integrins, inhibited the collagen-induced MT1-MMP-dependent activation of pro-MMP-2 and up-regulation of MT1-MMP at the gene and protein levels. Interestingly, DDR2 knockdown or pharmacological inhibition of DDR2 also inhibited the MT1-MMP-dependent cellular degradation of collagen film, suggesting that cell-surface collagen degradation by MT1-MMP involves DDR2-mediated collagen signaling. This DDR2-mediated mechanism is only present in non-transformed mesenchymal cells as collagen-induced MT1-MMP activation in HT1080 fibrosarcoma cells and MT1-MMP function in MDA-MB231 breast cancer cells were not affected by DDR kinase inhibition. DDR2 activation was found to be noticeably more effective when cells were stimulated by collagen without the non-helical telopeptide region compared with intact collagen fibrils. Furthermore, DDR2-dependent MT1-MMP activation by cartilage was found to be more efficient when the tissue was partially damaged. These data suggest that DDR2 is a microenvironment sensor that regulates fibroblast migration in a collagen-rich environment.

Footnotes

This work was supported by Arthritis Research UK Program Grant 19797 (to Y. I.), Arthritis Research UK Centre for Osteoarthritis Pathogenesis Grant 20205 (Y. I.), National Institutes of Health P01 Grant CA154303-04 (to N. S. G.), and a Ph.D. studentship from the Kennedy Trust for Rheumatology Research (to I. M. and Y. I.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.