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The protein tyrosine phosphatase PTPRJ/DEP-1 contributes to the regulation of the Notch-signaling pathway and sprouting angiogenesis

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Abstract

The Dll4-Notch-signaling pathway regulates capillary sprouting via the specification of endothelial tip cells. While VEGF is a potent inducer of Dll4 expression, the intracellular mediators that stimulate its expression remain poorly defined. The protein tyrosine phosphatase PTPRJ/DEP-1 is required for angiogenesis in normal or pathological contexts through its modulation of VEGF signaling. Here, we show that in DEP-1 KO mice, retinas at post-natal day 5 show enlarged blood vessels, as well as an increased number of tip cells and vessel branching points at the migrating front of the vascular plexus. Consistent with these observations, the proliferation of endothelial cells is increased in the retinas of DEP-1 KO mice, as revealed by phospho-histone H3 staining, and increased phosphorylation of ERK1/2 in HUVECs transfected with DEP-1 siRNA. The expression of Dll4 was decreased in retinas of DEP-1 KO mice and was associated with decreased Notch activation. Mechanistically, reduced Dll4 expression in the absence of DEP-1 was correlated with the inhibition of the Src/Akt/β-Catenin-signaling pathway in HUVECs. Conversely, overexpression of WT DEP-1 in cultured endothelial cells, but not of mutants unable to activate Src-dependent signaling, promoted Dll4 expression. Inhibition of Src, Akt, and β-catenin transcriptional activity, leading to the inhibition of Dll4 expression, further suggested that their activation through a DEP-1-dependent pathway was required to promote Dll4 expression in VEGF-stimulated endothelial cells. Altogether, these data demonstrate that DEP-1, via Akt and β-catenin, is a significant promoter of the VEGF-induced Dll4-Notch pathway, and can contribute to the regulation of the tip and stalk cell phenotypes of endothelial cells.

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Acknowledgements

We would like to thank A. Fusco for providing DEP-1 KO mice. This work was supported by operating grants from the Cancer Research Society (I.R. and B.L.), an operating grant from the Canadian Institutes of Health Research (363450) (B.L.) and a Grant-in-Aid from the Heart and Stroke Foundation of Canada (B.L.). B.L. is the recipient of a New Investigator Award from the Heart and Stroke Foundation of Canada. P. Fournier was supported by student scholarships from the Canadian Institutes of Health Research (292353) and Institut du cancer de Montréal.

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  1. Bruno Larrivée and Isabelle Royal contributed equally.

Authors and Affiliations

  1. CRCHUM - Centre de recherche du Centre Hospitalier de l’Université de Montréal, Montréal, QC, Canada

    Patrick Fournier & Isabelle Royal

  2. Institut du Cancer de Montréal, Montréal, QC, Canada

    Patrick Fournier & Isabelle Royal

  3. Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, Canada

    Claire Viallard, Agnieszka Dejda, Przemyslaw Sapieha & Bruno Larrivée

  4. Department of Ophthalmology, University of Montreal, Montréal, QC, Canada

    Przemyslaw Sapieha & Bruno Larrivée

  5. Department of Biomedical Sciences, University of Montreal, Montréal, QC, Canada

    Bruno Larrivée

  6. Department of Medicine, University of Montreal, Montréal, QC, Canada

    Isabelle Royal

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Fournier, P., Viallard, C., Dejda, A. et al. The protein tyrosine phosphatase PTPRJ/DEP-1 contributes to the regulation of the Notch-signaling pathway and sprouting angiogenesis. Angiogenesis 23, 145–157 (2020). https://doi.org/10.1007/s10456-019-09683-z

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