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

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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.

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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.

Author information

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.

Correspondence to Cédric Blanpain.

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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) doi:10.1038/nature10525

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