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Excess vascular endothelial growth factor-A disrupts pericyte recruitment during blood vessel formation

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Abstract

Pericyte investment into new blood vessels is essential for vascular development such that mis-regulation within this phase of vessel formation can contribute to numerous pathologies including arteriovenous and cerebrovascular malformations. It is critical therefore to illuminate how angiogenic signaling pathways intersect to regulate pericyte migration and investment. Here, we disrupted vascular endothelial growth factor-A (VEGF-A) signaling in ex vivo and in vitro models of sprouting angiogenesis, and found pericyte coverage to be compromised during VEGF-A perturbations. Pericytes had little to no expression of VEGF receptors, suggesting VEGF-A signaling defects affect endothelial cells directly but pericytes indirectly. Live imaging of ex vivo angiogenesis in mouse embryonic skin revealed limited pericyte migration during exposure to exogenous VEGF-A. During VEGF-A gain-of-function conditions, pericytes and endothelial cells displayed abnormal transcriptional changes within the platelet-derived growth factor-B (PDGF-B) and Notch pathways. To further test potential crosstalk between these pathways in pericytes, we stimulated embryonic pericytes with Notch ligands Delta-like 4 (Dll4) and Jagged-1 (Jag1) and found induction of Notch pathway activity but no changes in PDGF Receptor-β (Pdgfrβ) expression. In contrast, PDGFRβ protein levels decreased with mis-regulated VEGF-A activity, observed in the effects on full-length PDGFRβ and a truncated PDGFRβ isoform generated by proteolytic cleavage or potentially by mRNA splicing. Overall, these observations support a model in which, during the initial stages of vascular development, pericyte distribution and coverage are indirectly affected by endothelial cell VEGF-A signaling and the downstream regulation of PDGF-B-PDGFRβ dynamics, without substantial involvement of pericyte Notch signaling during these early stages.

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Abbreviations

Ng2:

Neural glial antigen-2

VEGF:

Vascular endothelial growth factor

ESC:

Embryonic stem cell

PDGF:

Platelet-derived growth factor

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Acknowledgements

We thank the Chappell Lab for critical and extensive discussions of the primary data.

Funding

This study was funded by the National Institutes of Health (R00HL105779 and R56HL133826 to JCC), National Heart, Lung, and Blood Institute (Grant Nos. HL105779, HL133826), Division of Chemical, Bioengineering, Environmental, and Transport Systems (Grant No. 1752339).

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Authors and Affiliations

  1. Center for Heart and Regenerative Medicine, Virginia Tech Carilion Research Institute, 2 Riverside Circle, Roanoke, VA, 24016, USA

    Jordan Darden, Laura Beth Payne, Huaning Zhao & John C. Chappell

  2. Graduate Program in Translational Biology, Medicine, and Health, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA

    Jordan Darden & John C. Chappell

  3. Department of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA

    Huaning Zhao & John C. Chappell

  4. Department of Basic Science Education, Virginia Tech Carilion School of Medicine, Roanoke, VA, 24016, USA

    John C. Chappell

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  1. Jordan Darden
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Correspondence to John C. Chappell.

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Darden, J., Payne, L.B., Zhao, H. et al. Excess vascular endothelial growth factor-A disrupts pericyte recruitment during blood vessel formation. Angiogenesis 22, 167–183 (2019). https://doi.org/10.1007/s10456-018-9648-z

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