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Defining the mode of tumour growth by clonal analysis

Nature volume 488, pages 527–530 (2012)Cite this article

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Abstract

Recent studies using the isolation of a subpopulation of tumour cells followed by their transplantation into immunodeficient mice provide evidence that certain tumours1,2, including squamous skin tumours3,4,5, contain cells with high clonogenic potential that have been referred to as cancer stem cells (CSCs). Until now, CSC properties have only been investigated by transplantation assays, and their existence in unperturbed tumour growth is unproven. Here we make use of clonal analysis of squamous skin tumours using genetic lineage tracing to unravel the mode of tumour growth in vivo in its native environment. To this end, we used a genetic labelling strategy that allows individual tumour cells to be marked and traced over time at different stages of tumour progression. Surprisingly, we found that the majority of labelled tumour cells in benign papilloma have only limited proliferative potential, whereas a fraction has the capacity to persist long term, giving rise to progeny that occupy a significant part of the tumour. As well as confirming the presence of two distinct proliferative cell compartments within the papilloma, mirroring the composition, hierarchy and fate behaviour of normal tissue, quantitative analysis of clonal fate data indicates that the more persistent population has stem-cell-like characteristics and cycles twice per day, whereas the second represents a slower cycling transient population that gives rise to terminally differentiated tumour cells. Such behaviour is shown to be consistent with double-labelling experiments and detailed clonal fate characteristics. By contrast, measurements of clone size and proliferative potential in invasive squamous cell carcinoma show a different pattern of behaviour, consistent with geometric expansion of a single CSC population with limited potential for terminal differentiation. This study presents the first experimental evidence for the existence of CSCs during unperturbed solid tumour growth.

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Figure 1: Genetic tracing of skin papilloma at clonal density.
Figure 2: Quantitative clonal analysis in skin papilloma.
Figure 3: Challenging the stem/progenitor cell model in papilloma.
Figure 4: Quantitative clonal analysis of squamous cell carcinoma.

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References

  1. Shackleton, M., Quintana, E., Fearon, E. R. & Morrison, S. J. Heterogeneity in cancer: cancer stem cells versus clonal evolution. Cell 138, 822–829 (2009)

    Article  CAS  Google Scholar 

  2. Lobo, N. A., Shimono, Y., Qian, D. & Clarke, M. F. The biology of cancer stem cells. Annu. Rev. Cell. Dev. Biol. 23, 675–699 (2007)

    Article  CAS  Google Scholar 

  3. Malanchi, I. et al. Cutaneous cancer stem cell maintenance is dependent on β-catenin signalling. Nature 452, 650–653 (2008)

    Article  CAS  ADS  Google Scholar 

  4. Beck, B. et al. A vascular niche and a VEGF–Nrp1 loop regulate the initiation and stemness of skin tumours. Nature 478, 399–403 (2011)

    Article  CAS  ADS  Google Scholar 

  5. Schober, M. & Fuchs, E. Tumor-initiating stem cells of squamous cell carcinomas and their control by TGF-β and integrin/focal adhesion kinase (FAK) signaling. Proc. Natl Acad. Sci. USA 108, 10544–10549 (2011)

    Article  CAS  ADS  Google Scholar 

  6. Nowell, P. C. The clonal evolution of tumor cell populations. Science 194, 23–28 (1976)

    Article  CAS  ADS  Google Scholar 

  7. Greaves, M. & Maley, C. C. Clonal evolution in cancer. Nature 481, 306–313 (2012)

    Article  CAS  ADS  Google Scholar 

  8. Nguyen, L. V., Vanner, R., Dirks, P. & Eaves, C. J. Cancer stem cells: an evolving concept. Nature Rev. Cancer 12, 133–143 (2012)

    Article  CAS  Google Scholar 

  9. Kemp, C. J. Multistep skin cancer in mice as a model to study the evolution of cancer cells. Semin. Cancer Biol. 15, 460–473 (2005)

    Article  CAS  Google Scholar 

  10. Abel, E. L., Angel, J. M., Kiguchi, K. & DiGiovanni, J. Multi-stage chemical carcinogenesis in mouse skin: fundamentals and applications. Nature Protocols 4, 1350–1362 (2009)

    Article  CAS  Google Scholar 

  11. Owens, D. M. & Watt, F. M. Contribution of stem cells and differentiated cells to epidermal tumours. Nature Rev. Cancer 3, 444–451 (2003)

    Article  CAS  Google Scholar 

  12. Perez-Losada, J. & Balmain, A. Stem-cell hierarchy in skin cancer. Nature Rev. Cancer 3, 434–443 (2003)

    Article  CAS  Google Scholar 

  13. Klein, C. A. Parallel progression of primary tumours and metastases. Nature Rev. Cancer 9, 302–312 (2009)

    Article  CAS  Google Scholar 

  14. Clayton, E. et al. A single type of progenitor cell maintains normal epidermis. Nature 446, 185–189 (2007)

    Article  CAS  ADS  Google Scholar 

  15. Doupé, D. P., Klein, A. M., Simons, B. D. & Jones, P. H. The ordered architecture of murine ear epidermis is maintained by progenitor cells with random fate. Dev. Cell 18, 317–323 (2010)

    Article  Google Scholar 

  16. Lopez-Garcia, C., Klein, A. M., Simons, B. D. & Winton, D. J. Intestinal stem cell replacement follows a pattern of neutral drift. Science 330, 822–825 (2010)

    Article  CAS  ADS  Google Scholar 

  17. Snippert, H. J. et al. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143, 134–144 (2010)

    Article  CAS  Google Scholar 

  18. Simons, B. D. & Clevers, H. Strategies for homeostatic stem cell self-renewal in adult tissues. Cell 145, 851–862 (2011)

    Article  CAS  Google Scholar 

  19. Klein, A. M. & Simons, B. D. Universal patterns of stem cell fate in cycling adult tissues. Development 138, 3103–3111 (2011)

    Article  CAS  Google Scholar 

  20. Ghazizadeh, S. & Taichman, L. B. Multiple classes of stem cells in cutaneous epithelium: a lineage analysis of adult mouse skin. EMBO J. 20, 1215–1222 (2001)

    Article  CAS  Google Scholar 

  21. Potten, C. S. Cell replacement in epidermis (keratopoiesis) via discrete units of proliferation. Int. Rev. Cytol. 69, 271–318 (1981)

    Article  CAS  Google Scholar 

  22. Mackenzie, I. C. Retroviral transduction of murine epidermal stem cells demonstrates clonal units of epidermal structure. J. Invest. Dermatol. 109, 377–383 (1997)

    Article  CAS  Google Scholar 

  23. Marusyk, A. & Polyak, K. Tumor heterogeneity: causes and consequences. Biochim. Biophys. Acta 1805, 105–117 (2010)

    CAS  PubMed  Google Scholar 

  24. Van Keymeulen, A. et al. Epidermal progenitors give rise to Merkel cells during embryonic development and adult homeostasis. J. Cell Biol. 187, 91–100 (2009)

    Article  CAS  Google Scholar 

  25. Rocheteau, P., Gayraud-Morel, B., Siegl-Cachedenier, I., Blasco, M. A. & Tajbakhsh, S. A subpopulation of adult skeletal muscle stem cells retains all template DNA strands after cell division. Cell 148, 112–125 (2012)

    Article  CAS  Google Scholar 

  26. Vasioukhin, V., Degenstein, L., Wise, B. & Fuchs, E. The magical touch: genome targeting in epidermal stem cells induced by tamoxifen application to mouse skin. Proc. Natl Acad. Sci. USA 96, 8551–8556 (1999)

    Article  CAS  ADS  Google Scholar 

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Acknowledgements

C.B. is investigator of WELBIO and chercheur qualifié of the FRS/FNRS. B.B. is chargé de recherche of the FRS/FNRS and G.D. is supported by the Brussels Region. This work was supported by the FNRS, the program d’excellence CIBLES of the Wallonia Region, a research grant from the Fondation Contre le Cancer, the ULB foundation and the fond Gaston Ithier, a starting grant of the European Research Council (ERC), and the EMBO Young Investigator Program. We thank F. Bollet-Quivogne and J.-M. Vanderwinden for their help with confocal imaging.

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Authors and Affiliations

  1. Université Libre de Bruxelles, IRIBHM, Brussels B-1070, Belgium,

    Gregory Driessens, Benjamin Beck, Amélie Caauwe & Cédric Blanpain

  2. Department of Physics, Cavendish Laboratory, J. J. Thomson Avenue, Cambridge CB3 0HE, UK,

    Benjamin D. Simons

  3. The Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK,

    Benjamin D. Simons

  4. WELBIO, Brussels B-1070, Belgium,

    Cédric Blanpain

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  1. Gregory Driessens
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  4. Benjamin D. Simons
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Contributions

C.B., G.D., B.B. and B.D.S. designed the experiments and performed data analysis. G.D. and B.B. performed most of the experiments. A.C. provided technical support. C.B. and B.D.S. wrote the manuscript.

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Correspondence to Benjamin D. Simons or Cédric Blanpain.

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The authors declare no competing financial interests.

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Driessens, G., Beck, B., Caauwe, A. et al. Defining the mode of tumour growth by clonal analysis. Nature 488, 527–530 (2012). https://doi.org/10.1038/nature11344

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