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A vascular niche and a VEGF–Nrp1 loop regulate the initiation and stemness of skin tumours

Abstract

Angiogenesis is critical during tumour initiation and malignant progression1. Different strategies aimed at blocking vascular endothelial growth factor (VEGF) and its receptors have been developed to inhibit angiogenesis in cancer patients2. It has become increasingly clear that in addition to its effect on angiogenesis, other mechanisms including a direct effect of VEGF on tumour cells may account for the efficiency of VEGF-blockade therapies3. Cancer stem cells (CSCs) have been described in various cancers including squamous tumours of the skin4,5. Here we use a mouse model of skin tumours to investigate the impact of the vascular niche and VEGF signalling on controlling the stemness (the ability to self renew and differentiate) of squamous skin tumours during the early stages of tumour progression. We show that CSCs of skin papillomas are localized in a perivascular niche, in the immediate vicinity of endothelial cells. Furthermore, blocking VEGFR2 caused tumour regression not only by decreasing the microvascular density, but also by reducing CSC pool size and impairing CSC renewal properties. Conditional deletion of Vegfa in tumour epithelial cells caused tumours to regress, whereas VEGF overexpression by tumour epithelial cells accelerated tumour growth. In addition to its well-known effect on angiogenesis, VEGF affected skin tumour growth by promoting cancer stemness and symmetric CSC division, leading to CSC expansion. Moreover, deletion of neuropilin-1 (Nrp1), a VEGF co-receptor expressed in cutaneous CSCs, blocked VEGF’s ability to promote cancer stemness and renewal. Our results identify a dual role for tumour-cell-derived VEGF in promoting cancer stemness: by stimulating angiogenesis in a paracrine manner, VEGF creates a perivascular niche for CSCs, and by directly affecting CSCs through Nrp1 in an autocrine loop, VEGF stimulates cancer stemness and renewal. Finally, deletion of Nrp1 in normal epidermis prevents skin tumour initiation. These results may have important implications for the prevention and treatment of skin cancers.

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Figure 1: A perivascular niche regulates cancer stemness in skin papilloma.
Figure 2: VEGF expression by tumour cells is required for the maintenance of cutaneous cancer stem cells.
Figure 3: Increased VEGF expression promotes cutaneous cancer-stem-cell expansion.
Figure 4: Nrp1 is required for squamous tumour initiation and regulates stemness of cutaneous cancer cells.

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Gene Expression Omnibus

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Microarray data have been deposited in the GEO database under accession number GSE31465.

References

  1. 1

    Kerbel, R. S. Tumor angiogenesis. N. Engl. J. Med. 358, 2039–2049 (2008)

    CAS  Article  Google Scholar 

  2. 2

    Ferrara, N., Mass, R. D., Campa, C. & Kim, R. Targeting VEGF-A to treat cancer and age-related macular degeneration. Annu. Rev. Med. 58, 491–504 (2007)

    CAS  Article  Google Scholar 

  3. 3

    Ellis, L. M. & Hicklin, D. J. VEGF-targeted therapy: mechanisms of anti-tumour activity. Nature Rev. Cancer 8, 579–591 (2008)

    CAS  Article  Google Scholar 

  4. 4

    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)

    CAS  Article  Google Scholar 

  5. 5

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

    CAS  Article  ADS  Google Scholar 

  6. 6

    Alam, M. & Ratner, D. Cutaneous squamous-cell carcinoma. N. Engl. J. Med. 344, 975–983 (2001)

    CAS  Article  Google Scholar 

  7. 7

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

    CAS  Article  Google Scholar 

  8. 8

    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)

    CAS  Article  Google Scholar 

  9. 9

    Calabrese, C. et al. A perivascular niche for brain tumor stem cells. Cancer Cell 11, 69–82 (2007)

    CAS  Article  Google Scholar 

  10. 10

    Butler, J. M., Kobayashi, H. & Rafii, S. Instructive role of the vascular niche in promoting tumour growth and tissue repair by angiocrine factors. Nature Rev. Cancer 10, 138–146 (2010)

    CAS  Article  Google Scholar 

  11. 11

    Kiel, M. J. & Morrison, S. J. Maintaining hematopoietic stem cells in the vascular niche. Immunity 25, 862–864 (2006)

    CAS  Article  Google Scholar 

  12. 12

    Witte, L. et al. Monoclonal antibodies targeting the VEGF receptor-2 (Flk1/KDR) as an anti-angiogenic therapeutic strategy. Cancer Metastasis Rev. 17, 155–161 (1998)

    CAS  Article  Google Scholar 

  13. 13

    Miller, D. W. et al. Rapid vessel regression, protease inhibition, and stromal normalization upon short-term vascular endothelial growth factor receptor 2 inhibition in skin carcinoma heterotransplants. Am. J. Pathol. 167, 1389–1403 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Lichtenberger, B. M. et al. Autocrine VEGF signaling synergizes with EGFR in tumor cells to promote epithelial cancer development. Cell 140, 268–279 (2010)

    CAS  Article  Google Scholar 

  15. 15

    Rossiter, H. et al. Loss of vascular endothelial growth factor A activity in murine epidermal keratinocytes delays wound healing and inhibits tumor formation. Cancer Res. 64, 3508–3516 (2004)

    CAS  Article  Google Scholar 

  16. 16

    Hirakawa, S. et al. VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis. J. Exp. Med. 201, 1089–1099 (2005)

    CAS  Article  Google Scholar 

  17. 17

    Larcher, F., Murillas, R., Bolontrade, M., Conti, C. J. & Jorcano, J. L. VEGF/VPF overexpression in skin of transgenic mice induces angiogenesis, vascular hyperpermeability and accelerated tumor development. Oncogene 17, 303–311 (1998)

    CAS  Article  Google Scholar 

  18. 18

    Gerber, H. P. et al. VEGF is required for growth and survival in neonatal mice. Development 126, 1149–1159 (1999)

    CAS  PubMed  Google Scholar 

  19. 19

    Lechler, T. & Fuchs, E. Asymmetric cell divisions promote stratification and differentiation of mammalian skin. Nature 437, 275–280 (2005)

    CAS  Article  ADS  Google Scholar 

  20. 20

    Poulson, N. D. & Lechler, T. Robust control of mitotic spindle orientation in the developing epidermis. J. Cell Biol. 191, 915–922 (2010)

    CAS  Article  Google Scholar 

  21. 21

    Williams, S. E., Beronja, S., Pasolli, H. A. & Fuchs, E. Asymmetric cell divisions promote Notch-dependent epidermal differentiation. Nature 470, 353–358 (2011)

    CAS  Article  ADS  Google Scholar 

  22. 22

    Maes, C. et al. Increased skeletal VEGF enhances β-catenin activity and results in excessively ossified bones. EMBO J. 29, 424–441 (2010)

    CAS  Article  Google Scholar 

  23. 23

    Soker, S., Takashima, S., Miao, H. Q., Neufeld, G. & Klagsbrun, M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92, 735–745 (1998)

    CAS  Article  Google Scholar 

  24. 24

    Staton, C. A., Kumar, I., Reed, M. W. & Brown, N. J. Neuropilins in physiological and pathological angiogenesis. J. Pathol. 212, 237–248 (2007)

    CAS  Article  Google Scholar 

  25. 25

    Pan, Q. et al. Blocking neuropilin-1 function has an additive effect with anti-VEGF to inhibit tumor growth. Cancer Cell 11, 53–67 (2007)

    CAS  Article  Google Scholar 

  26. 26

    Vasioukhin, V., Bauer, C., Degenstein, L., Wise, B. & Fuchs, E. Hyperproliferation and defects in epithelial polarity upon conditional ablation of alpha-catenin in skin. Cell 104, 605–617 (2001)

    CAS  Article  Google Scholar 

  27. 27

    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)

    CAS  Article  ADS  Google Scholar 

  28. 28

    Gerber, H. P. et al. VEGF is required for growth and survival in neonatal mice. Development 126, 1149–1159 (1999)

    CAS  PubMed  Google Scholar 

  29. 29

    Gu, C. et al. Neuropilin-1 conveys semaphorin and VEGF signaling during neural and cardiovascular development. Dev. Cell 5, 45–57 (2003)

    CAS  Article  Google Scholar 

  30. 30

    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)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank Genetech for providing blocking anti-Nrp1 antibodies and H. Fujisawa, N. Ferrara and A. Nagy for providing the Nrp1fl/−, Vegffl/fl and ROSA26-VEGF-164 mice, respectively. C.B. is an investigator of Welbio. C.B. and P.A.S. are chercheur qualifié and B.B. is a chargé de recherche of the FRS/FNRS; Am.C. and G.M. are research fellows of the FRS/FRIA. G.D. is supported by the Brussels Region, B.D. and K.K.Y. by TELEVIE and. S.G. is a postdoctoral fellow of the Basic Science Research Foundation-Flanders (FWO). S.L. is funded by the Max-Eder group leader program of the Deutsche Krebshilfe, by the Hamburger Krebsgesellschaft and by the Roggenbuck Stiftung. P.C. is funded by Long-term structural Methusalem funding by the Flemish Government. 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.

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C.B., B.B., G.D., S.G., K.K.Y., P.C. and J.J.H. designed the experiments and performed data analysis. B.B., G.D., S.G., K.K.Y., G.L., S.L. and A.K. performed most of the experiments; P.A.S., Au.C., G.M. and B.D. contributed to mice treatment; S.D. and Am.C. provided technical support. C.B. and B.B. wrote the manuscript.

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Correspondence to Cédric Blanpain.

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The authors declare no competing financial interests.

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Beck, B., Driessens, G., Goossens, S. et al. A vascular niche and a VEGF–Nrp1 loop regulate the initiation and stemness of skin tumours. Nature 478, 399–403 (2011). https://doi.org/10.1038/nature10525

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