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A mechanically active heterotypic E-cadherin/N-cadherin adhesion enables fibroblasts to drive cancer cell invasion

Nature Cell Biology volume 19, pages 224237 (2017) | Download Citation


Cancer-associated fibroblasts (CAFs) promote tumour invasion and metastasis. We show that CAFs exert a physical force on cancer cells that enables their collective invasion. Force transmission is mediated by a heterophilic adhesion involving N-cadherin at the CAF membrane and E-cadherin at the cancer cell membrane. This adhesion is mechanically active; when subjected to force it triggers β-catenin recruitment and adhesion reinforcement dependent on α-catenin/vinculin interaction. Impairment of E-cadherin/N-cadherin adhesion abrogates the ability of CAFs to guide collective cell migration and blocks cancer cell invasion. N-cadherin also mediates repolarization of the CAFs away from the cancer cells. In parallel, nectins and afadin are recruited to the cancer cell/CAF interface and CAF repolarization is afadin dependent. Heterotypic junctions between CAFs and cancer cells are observed in patient-derived material. Together, our findings show that a mechanically active heterophilic adhesion between CAFs and cancer cells enables cooperative tumour invasion.

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We thank N. Castro for technical assistance, J. de Rooij (UMC Utrecht, Netherlands) for plasmids, S. Pérez-Amodio (IBEC, Spain) for dermal fibroblasts, N. Reguart (Hospital Clinic, Spain) and M. Gabasa (University of Barcelona, Spain) for lung fibroblasts, and A. Schertel (Zeiss) for assistance with the FIB-SEM. This work was supported by the Spanish Ministry of Economy and Competitiveness/FEDER (BFU2012-38146 to X.T., BFU2014-52586-REDT to P.R.-C., IJCI2014-19843 to A.L. and IJCI-2014-19156 to A.E.-A.), the Generalitat de Catalunya (2014-SGR-927 to X.T. and CERCA Programme), the European Research Council (StG-CoG-616480 to X.T.), Obra Social ‘La Caixa’, Marie-Curie action (CAFFORCE 328664 to A.L.), EMBO Long-term fellowship (EMBO ALTF 1235-2012 to A.L.), a Career Integration Grant within the seventh European Community Framework Programme (PCIG10-GA-2011-303848 to P.R.-C.), Fundació la Marató de TV3 (project 20133330 to P.R.-C.), and AXA research fund (L.A.). E.S., E.A., A.W. and S.D. are funded by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001144), the UK Medical Research Council (FC001144), and the Wellcome Trust (FC001144). T.K. is funded by Marie-Curie action (HeteroCancerInvasion no. 708651) and the Japanese Strategic Young Researcher Overseas Visits Program for Accelerating Brain Circulation.

Author information


  1. Institute for Bioengineering of Catalonia, Barcelona 08028, Spain

    • Anna Labernadie
    • , Agustí Brugués
    • , Xavier Serra-Picamal
    • , Victor González-Tarragó
    • , Alberto Elosegui-Artola
    • , Lorenzo Albertazzi
    • , Pere Roca-Cusachs
    •  & Xavier Trepat
  2. The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK

    • Takuya Kato
    • , Stefanie Derzsi
    • , Esther Arwert
    • , Anne Weston
    •  & Erik Sahai
  3. Unitat de Biofísica i Bioenginyeria, Facultat de Medicina, Universitat de Barcelona, Barcelona 08036, Spain

    • Xavier Serra-Picamal
    • , Jordi Alcaraz
    • , Pere Roca-Cusachs
    •  & Xavier Trepat
  4. Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona 08010, Spain

    • Xavier Trepat
  5. Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina, Barcelona 08028, Spain

    • Xavier Trepat


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A.L., E.S. and X.T. conceived the study and designed experiments, with additional input from T.K. A.L. performed most experiments and data analysis. T.K. performed and analysed spheroid invasion experiments and generated A431 KO cell lines. A.L., A.E.-A., V.G.-T. and P.R.-C. designed, performed and analysed magnetic cytometry assays. A.B. and X.S.-P. developed software for image analysis and force measurements. S.D. performed QRT–PCR experiments. A.L. and L.A. performed STORM imaging. J.A. contributed CAFs from patients with non-small lung cell carcinoma. E.A. performed the intravital imaging and assisted with the patient sample analysis, A.W. performed electron microscopy. A.L., E.S. and X.T. wrote the manuscript with input from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Erik Sahai or Xavier Trepat.

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  1. 1.

    CAFs lead cancer cell strands in 3D invasion assays.

    Representative 3D rendering of a fixed spheroid containing vulval CAFs (VCAF, CAGAP-cherry) and A431 cells (green) (ratio 1:1) after 24 h embedded in an organotypic ECM. z-step, 0.25 μm.

  2. 2.

    CAFs favor expansion of cancer cell spheroids in 2D.

    Representative time-lapse of a 2D migration assay on a soft polyacrylamide substrate (Young’s modulus E = 6 kPa). White rectangles highlight three CAFs (CAGAP-mCherry) leading the expansion of the A431 spheroid (unlabeled). Images were acquired every 5 min. Scale bar, 100 μm.

  3. 3.

    CAFs lead collective migration of cancer cells in 2D.

    Representative time-lapse of one CAF (CAGAP-mCherry) leading the 2D migration of A431 cells (unlabeled) away from the spheroid edge. Cells are adhered on an elastic substrate (Young’s modulus E = 6 kPa). Images were acquired every 5 min. Scale bar, 2 μm.

  4. 4.

    FIB-SEM reveals multiple contact points at the CAF-A431 interface.

    Representative FIB-SEM z-stack sequence of areas of contact between CAFs (VCAF) and A431 cells. White arrows show the location of contact between CAF and cancer cells. Image dimensions, 10 × 12 μm, z-stack steps 50 nm.

  5. 5.

    W2A mutation in the extracellular domain of E-cadherin drastically diminishes co-localization with N-cadherin.

    Representative confocal time-lapse movie of A431-EcadWT-Ruby or A431-EcadW2A-Ruby cells co-cultured with CAFs expressing N-cadherin-GFP. Dynamics of the co-culture was recorded at 5 min intervals. Scale Bar, 20  μm.

  6. 6.

    Calcium chelation abrogates reversibly E-cadherin/N-cadherin co-localization.

    Representative confocal time-lapse movie of A431-E-cadherin-Ruby mixed with CAF-N-cadherin-GFP showing dynamics of the E-cadherin/N-cadherin contact during a calcium chelation assay. After 10 min of acquisition, the EGTA solution was added to the medium (final concentration, 4 mM). After 4 min incubation, the medium containing EGTA was washed three times with normal medium. Arrows show the formation of the E-cadherin/N-cadherin contact after washout of EGTA. Images were acquired every 2 min. Scale Bar, 20 μm.

  7. 7.

    Dynamics of the E-cadherin/N-cadherin adhesion during CAF-led cancer cell migration.

    Representative confocal time-lapse movie of A431-E-cadherin-Ruby spheroid seeded on glass and surrounded by CAF-N-cadherin-GFP. The magnified panel represents the area of contact between the leading CAF and A431 cells. White arrow shows the location of the E-cadherin/N-cadherin contact. Images were acquired every 5 min. Scale Bars, 20 μm (right panel), 10 μm (left panel).

  8. 8.

    E-cadherin and β-catenin colocalize at heterotypic contacts.

    Representative confocal time-lapse movie of A431-E-cadherin-Ruby expressing β-catenin-GFP mixed with unlabeled CAFs. The magnified panel represents the area of contact between the leading CAF and the A431 cells. The white arrow shows the localization of the contact between the A431 cells and the CAF (white asterisk). Images were acquired every 2 min. Scale Bar, 10 μm.

  9. 9.

    E-cadherin and vinculin colocalize at heterotypic contacts.

    Representative confocal time-lapse movie of A431-E-cadherin-Ruby expressing vinculin-GFP mixed with unlabeled CAFs. The white arrow shows the localization of the contact between the A431 cells and the CAF (white asterisk). Note an enrichment of E-cadherin (red) and vinculin (green) at the contact. Images were acquired every 2 min. Scale Bar, 10 μm.

  10. 10.

    CAFs exert pulling forces on cancer cells.

    Representative time-lapse of a CAF (CAGAP-mCherry) dragging A431 cells. The magnitude and direction of the force exerted by the CAF on the cancer cell is represented by the green vector. For clarity, the force vector is represented at the geometric center of the CAF. See Fig. 5 for a quantification of the force throughout the time-lapse. Scale bar, 50 μm.

  11. 11.

    E-cadherin is required for force transmission between CAFs and A431 cells.

    Representative time-lapse of a CAF contacting the edge of A431-EcadKO cells. The magnitude and direction of the force exerted by the CAF on the cancer cell is represented by the green vector. For clarity, the force vector is represented at the geometric center of the CAF. See Fig. 5 for a quantification of the force throughout the time-lapse. Scale bar, 50 μm.

  12. 12.

    ‘Leader’ versus ‘loner’ CAF phenotypes.

    Representative time-lapse of a ‘leader’ CAF (left) and a ‘loner’ CAF (right) (CAGAP-mCherry, white arrow). Images were acquired every 5 min. Scale bars, 20 μm.

  13. 13.

    The heterotypic contact regulates CAF repolarization.

    Representative time-lapse of a control CAF (left panel) or N-cadherin depleted CAF (CAF-siNcad, right panel) contacting the edge of a spheroid of A431 control cells (left and right panel) or A431-EcadKO cells (middle panel). Colored spots show the location of the CAFs. Images were acquired every 10 min. Scale bar, 50 μm.

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