Abstract
Angiogenesis, the formation of new blood vessels, is fundamental to development and post-injury tissue repair. Vascular endothelial growth factor (VEGF)-A guides and enhances endothelial cell migration to initiate angiogenesis. Profilin-1 (Pfn-1) is an actin-binding protein that enhances actin filament formation and cell migration, but stimulus-dependent regulation of Pfn-1 has not been observed. Here, we show that VEGF-A-inducible phosphorylation of Pfn-1 at Tyr 129 is critical for endothelial cell migration and angiogenesis. Chemotactic activation of VEGF receptor kinase-2 (VEGFR2) and Src induces Pfn-1 phosphorylation in the cell leading edge, promoting Pfn-1 binding to actin and actin polymerization. Conditional endothelial knock-in of phosphorylation-deficient Pfn1Y129F in mice reveals that Pfn-1 phosphorylation is critical for angiogenesis in response to wounding and ischaemic injury, but not for developmental angiogenesis. Thus, VEGFR2/Src-mediated phosphorylation of Pfn-1 bypasses canonical, multistep intracellular signalling events to initiate endothelial cell migration and angiogenesis, and might serve as a selective therapeutic target for anti-angiogenic therapy.
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Acknowledgements
We are grateful to A. Levine and T. Egelhoff for helpful suggestions, X. West, A. Vasanji and J. Boerckel for technical assistance in punch-wound and microCT studies, and J. Drazba for image analysis. Cdh5-Cre/ERT2 mice were provided by R. Adams, Max Planck Institute, Germany. This work was supported by National Institutes of Health grants P01 HL029582, P01 HL076491 and R21 HL094841 (to P.L.F.).
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Y.F. designed, performed and analysed all experiments, produced figures, and wrote the initial draft of the paper. A.A. generated and analysed the data in Figs 1c–e, 2e and Supplementary Fig. S4d. Y.Q. designed and performed immunohistochemistry analysis in Fig. 5a. J.J. contributed to the analysis of the BIAcore in Fig. 2d. S.M.E. performed phylogenic analysis of Pfn-1. B.W. identified the Pfn-1 phosphorylation site by mass spectrometry as shown in Fig. 1b and Supplementary Fig. S2. A.H., L.M.G. and M.S.P. helped write and edit the final draft of the manuscript. P.L.F designed, supervised, and analysed all experiments and wrote the final draft of the manuscript.
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Fan, Y., Arif, A., Gong, Y. et al. Stimulus-dependent phosphorylation of profilin-1 in angiogenesis. Nat Cell Biol 14, 1046–1056 (2012). https://doi.org/10.1038/ncb2580
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DOI: https://doi.org/10.1038/ncb2580
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