Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published January 2018 | Published + Supplemental Material
Journal Article Open

Flow-Responsive Vascular Endothelial Growth Factor Receptor-Protein Kinase C Isoform Epsilon Signaling Mediates Glycolytic Metabolites for Vascular Repair


Aims: Hemodynamic shear stress participates in maintaining vascular redox status. Elucidating flow-mediated endothelial metabolites enables us to discover metabolic biomarkers and therapeutic targets. We posited that flow-responsive vascular endothelial growth factor receptor (VEGFR)-protein kinase C isoform epsilon (PKCɛ)-6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) signaling modulates glycolytic metabolites for vascular repair. Results: Bidirectional oscillatory flow (oscillatory shear stress [OSS]: 0.1 ± 3 dyne·cm^(−2) at 1 Hz) upregulated VEGFR-dependent PKCɛ expression to a greater degree than did unidirectional pulsatile flow (pulsatile shear stress [PSS]: 23 ± 8 dyne·cm^(−2) at 1 Hz) in human aortic endothelial cells (p < 0.05, n = 3). PSS and OSS further upregulated PKCɛ-dependent PFKFB3 expression for glycolysis (p < 0.05, n = 4). Constitutively active PKCɛ increased, whereas dominant-negative PKCɛ reduced both basal and maximal extracellular acidification rates for glycolytic flux (p < 0.01, n = 4). Metabolomic analysis demonstrated an increase in PKCɛ-dependent glycolytic metabolite, dihydroxyacetone (DHA), but a decrease in gluconeogenic metabolite, aspartic acid (p < 0.05 vs. control, n = 6). In a New Zealand White rabbit model, both PKCɛ and PFKFB3 immunostaining was prominent in the PSS- and OSS-exposed aortic arch and descending aorta. In a transgenic Tg(flk-1:EGFP) zebrafish model, GATA-1a morpholino oligonucleotide injection (to reduce viscosity-dependent shear stress) impaired vascular regeneration after tail amputation (p < 0.01, n = 20), which was restored with PKCɛ messenger RNA (mRNA) rescue (p < 0.05, n = 5). As a corollary, siPKCɛ inhibited tube formation and vascular repair, which were restored by DHA treatment in our Matrigel and zebrafish models. Innovation and Conclusion: Flow-sensitive VEGFR-PKCɛ-PFKFB3 signaling increases the glycolytic metabolite, dihydroxyacetone, to promote vascular repair.

Additional Information

© 2017 Mary Ann Liebert, Inc. publishers. Online Ahead of Print: September 21, 2017; Online Ahead of Editing: August 1, 2017. The authors would like to express their gratitude to Dr. Manuel Mayr for providing the metabolomic analysis of PKCɛ-mediated cardiac metabolites. Human CA-PKCɛ and dominant-negative PKCɛ adenoviruses were generously provided by Dr. Allen Samarel at the Loyola University Medical Center. This study was supported by the National Institutes of Health HL118650, HL083015, and HL129727 (T.K.H.). No competing financial interests exist.

Attached Files

Published - ars.2017.7044.pdf

Supplemental Material - Supp_Fig3.pdf

Supplemental Material - Supp_Fig4.pdf

Supplemental Material - Supp_Fig5.pdf

Supplemental Material - Supp_Fig6.pdf

Supplemental Material - Supp_Fig7.pdf

Supplemental Material - Supp_Figs1-2.pdf

Supplemental Material - Supp_Table1.zip

Supplemental Material - Supp_Video1.zip

Supplemental Material - Supp_Video2.zip

Supplemental Material - Supp_Video3.zip


Files (15.7 MB)
Name Size Download all
106.6 kB Preview Download
7.7 MB Preview Download
111.0 kB Preview Download
3.9 MB Preview Download
169.4 kB Preview Download
182.2 kB Preview Download
1.6 MB Preview Download
102.6 kB Preview Download
213.3 kB Preview Download
342.2 kB Preview Download
1.4 MB Preview Download

Additional details

August 19, 2023
October 17, 2023