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Wnt activity defines colon cancer stem cells and is regulated by the microenvironment

Nature Cell Biology volume 12, pages 468–476 (2010)Cite this article

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Abstract

Despite the presence of mutations in APC or β-catenin, which are believed to activate the Wnt signalling cascade constitutively, most colorectal cancers show cellular heterogeneity when β-catenin localization is analysed, indicating a more complex regulation of Wnt signalling. We explored this heterogeneity with a Wnt reporter construct and observed that high Wnt activity functionally designates the colon cancer stem cell (CSC) population. In adenocarcinomas, high activity of the Wnt pathway is observed preferentially in tumour cells located close to stromal myofibroblasts, indicating that Wnt activity and cancer stemness may be regulated by extrinsic cues. In agreement with this notion, myofibroblast-secreted factors, specifically hepatocyte growth factor, activate β-catenin-dependent transcription and subsequently CSC clonogenicity. More significantly, myofibroblast-secreted factors also restore the CSC phenotype in more differentiated tumour cells both in vitro and in vivo. We therefore propose that stemness of colon cancer cells is in part orchestrated by the microenvironment and is a much more dynamic quality than previously expected that can be defined by high Wnt activity.

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Figure 1: Wnt heterogeneity in primary tumours and in CSC culture.
Figure 2: Cells with high Wnt signalling show CSC properties in vitro.
Figure 3: Cells with high Wnt signalling show CSC properties in vivo.
Figure 4: Ex vivo tumorigenic assay and single-cell-derived tumours.
Figure 5: Myofibroblasts support stem-cell properties and regulate Wnt signalling.
Figure 6: Myofibroblasts restore Wnt activity and clonogenic potential in TOP–GFPlow cells.
Figure 7: Myofibroblasts restore tumorigenicity in TOP–GFPlow Wnt-active cells and co-localize with highly Wnt-active cells in vivo.
Figure 8: Schematic representation of the proposed model.

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Acknowledgements

The authors are indebted to the work performed by Willem Bemelman and Thomas van Gulik, surgeons at the AMC without whom the studies described here would not have been possible. In addition, we thank Riccardo Fodde for constructive discussions on CSCs and Wnt, and the animal care takers for their work for this project. Finally, we thank Berend Hooibrink and Toni van Capel for assistance with fluorescence-activated cell sorting experiments. This work was supported by a VICI grant from the Netherlands Organisation for Scientific Research and a Dutch Cancer Society (KWF Kankerbestrijding) grant (2009-4416) (to J.P.M.), an Academisch Medisch Centrum (AMC) fellowship (to L.V. and F.d.S.M.), the AMC Graduate School (to K.K.) and by the Associazione Italiana per la Ricerca sul Cancro (AIRC) (to G.S. and M.T.).

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Author notes

  1. Giorgio Stassi

    Present address: Present address: Cellular and Molecular Oncology, IRCCS Fondazione Salvatore Maugeri, 27100 Pavia, Italy.,

  2. Louis Vermeulen and Felipe De Sousa E Melo: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory for Experimental Oncology and Radiobiology (LEXOR), Center for Experimental Molecular Medicine (CEMM), Academic Medical Center (AMC), University of Amsterdam, 1105 AZ, Amsterdam, The Netherlands

    Louis Vermeulen, Felipe De Sousa E Melo, Maartje van der Heijden, Kate Cameron, Joan H. de Jong, Tijana Borovski, Jurriaan B. Tuynman, Hans Rodermond, Martin R. Sprick, Kristel Kemper, Dick J. Richel & Jan Paul Medema

  2. Department of Surgical and Oncological Sciences, Cellular and Molecular Pathophysiology Laboratory, University of Palermo, 90127, Palermo, Italy

    Matilde Todaro & Giorgio Stassi

  3. APOGENIX GmbH, 69120, Heidelberg, Germany

    Christian Merz

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Contributions

L.V., F.d.S.M., M.v.d.H., K.C., J.H.d.J., T.B., J.B.T., H.R., M.R.S., K.K. designed and conducted experiments. M.T. and C.M. isolated and cultured CSC lines, D.J.R., G.S. and J.P.M. planned and supervised the experiments. L.V., F.d.S.M and J.P.M wrote the manuscript.

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Correspondence to Jan Paul Medema.

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Vermeulen, L., De Sousa E Melo, F., van der Heijden, M. et al. Wnt activity defines colon cancer stem cells and is regulated by the microenvironment. Nat Cell Biol 12, 468–476 (2010). https://doi.org/10.1038/ncb2048

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