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Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion

Abstract

Invasive cell migration through tissue barriers requires pericellular remodelling of extracellular matrix (ECM) executed by cell-surface proteases, particularly membrane-type-1 matrix metalloproteinase (MT1-MMP/MMP-14). Using time-resolved multimodal microscopy, we show how invasive HT-1080 fibrosarcoma and MDA-MB-231 breast cancer cells coordinate mechanotransduction and fibrillar collagen remodelling by segregating the anterior force-generating leading edge containing β1 integrin, MT1-MMP and F-actin from a posterior proteolytic zone executing fibre breakdown. During forward movement, sterically impeding fibres are selectively realigned into microtracks of single-cell calibre. Microtracks become expanded by multiple following cells by means of the large-scale degradation of lateral ECM interfaces, ultimately prompting transition towards collective invasion similar to that in vivo. Both ECM track widening and transition to multicellular invasion are dependent on MT1-MMP-mediated collagenolysis, shown by broad-spectrum protease inhibition and RNA interference. Thus, invasive migration and proteolytic ECM remodelling are interdependent processes that control tissue micropatterning and macropatterning and, consequently, individual and collective cell migration.

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Figure 1: Enhanced collagenolysis and migration of HT-1080 cells in 3D collagen lattices after overexpression of MT1-MMP.
Figure 2: Distinct zones for adhesion, MT1-MMP location and collagenolysis in single migrating cells.
Figure 3: Real-time detection of collagen fibre cleavage and displacement.
Figure 4: Cell deformation after inhibition of pericellular collagenolysis.
Figure 5: Proteolytic and non-proteolytic migration in high-density collagen spheroid model.
Figure 6: Large-scale removal of ECM layers during multicellular invasion.
Figure 7: Requirement for pericellular collagenolysis in invasive multicellular but not single-cell migration.
Figure 8: Abrogation of collagenolytic track formation and collective invasion after knockdown of MT1-MMP in HT-MT1 cells.

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Acknowledgements

We thank M. Ott, M. Kuhn and A. Staudigel for excellent technical assistance; L. King for supplying COL2¾Cshort antibody; British Biotech Inc., UK, for supplying BB-2516; R. Fridman for supplying recombinant TIMP-1 and TIMP-2; E. Deryugina and A. Y. Strongin for providing HT-1080 cell lines; D. Pei for supplying human MT1-MMP cDNA; K. Müller-Hermelink for providing access to the confocal facility; and G. Krohne for providing access to electron microscopy. This work was supported by the Deutsche Forschungsgemeinschaft (FR 1155/7-1) and Deutsche Krebshilfe (AZ 106950). C.O. was supported by a Canada Research Chair Grant in Metalloproteinase Proteomics and Systems Biology, and E.T. by a CIHR Cell Signals Training Grant. S.S., Y.W. and Y.L. were supported by a research grant from NIH (RO1 CA86984).

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P.F. and K.W. conceived and designed the experiments. K.W., Y.W., Y.L. and E.T. performed the experiments. K.W., P.F. and J.G. analysed the data. S.S., J.G. and C.O. contributed reagents, material and analysis tools. K.W. and P.F. wrote the paper. All authors read and corrected the manuscript.

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Correspondence to Peter Friedl.

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Wolf, K., Wu, Y., Liu, Y. et al. Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion. Nat Cell Biol 9, 893–904 (2007). https://doi.org/10.1038/ncb1616

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