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Contribution of vascular endothelial growth factor receptor-2 sialylation to the process of angiogenesis

Oncogene volume 36, pages 6531–6541 (2017)Cite this article

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Abstract

Vascular endothelial growth factor receptor-2 (VEGFR2) is the main pro-angiogenic receptor expressed by endothelial cells (ECs). Using surface plasmon resonance, immunoprecipitation, enzymatic digestion, immunofluorescence and cross-linking experiments with specific sugar-binding lectins, we demonstrated that VEGFR2 bears both α,1-fucose and α(2,6)-linked sialic acid (NeuAc). However, only the latter is required for VEGF binding to VEGFR2 and consequent VEGF-dependent VEGFR2 activation and motogenic response in ECs. Notably, downregulation of β-galactoside α(2,6)-sialyltransferase expression by short hairpin RNA transduction inhibits VEGFR2 α(2,6) sialylation that is paralleled by an increase of β-galactoside α(2,3)-sialyltransferase expression. This results in an ex-novo α(2,3)-NeuAc sialylation of the receptor that functionally replaces the lacking α(2,6)-NeuAc, thus allowing VEGF/VEGFR2 interaction. In keeping with the role of VEGFR2 sialylation in angiogenesis, the α(2,6)-NeuAc-binding lectin Sambucus nigra (SNA) prevents VEGF-dependent VEGFR2 autophosphorylation and EC motility, proliferation and motogenesis. In addition, SNA exerts a VEGF-antagonist activity in tridimensional angiogenesis models in vitro and in the chick-embryo chorioallantoic membrane neovascularization assay and mouse matrigel plug assay in vivo. In conclusion, VEGFR2-associated NeuAc plays an important role in modulating VEGF/VEGFR2 interaction, EC pro-angiogenic activation and neovessel formation. VEGFR2 sialylation may represent a target for the treatment of angiogenesis-dependent diseases.

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Acknowledgements

This work was supported by grant from Ministero dell’Istruzione, Università e Ricerca (MIUR) (ex 60%) to MR and by IG 18943 and MFAG 18459 grants from Associazione Italiana per la Ricerca sul Cancro (AIRC) to MP and RR, respectively. PC and SR were supported by AIRC Fellowships.

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  1. Units of Experimental Oncology and Immunology, University of Brescia, Brescia, Italy

    P Chiodelli, S Rezzola, C Urbinati, F Federici Signori, R Ronca, M Presta & M Rusnati

  2. Department of Molecular and Translational Medicine, Units of Biotechnology, University of Brescia, Brescia, Italy

    E Monti

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  1. P Chiodelli
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Chiodelli, P., Rezzola, S., Urbinati, C. et al. Contribution of vascular endothelial growth factor receptor-2 sialylation to the process of angiogenesis. Oncogene 36, 6531–6541 (2017). https://doi.org/10.1038/onc.2017.243

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