NYMC Faculty Publications

20-HETE Regulates the Angiogenic Functions of Human Endothelial Progenitor Cells and Contributes to Angiogenesis in Vivo

Author Type(s)

Faculty

Additional Author Affiliation

Touro College

DOI

10.1124/jpet.113.210120

Journal Title

The Journal of Pharmacology and Experimental Therapeutics

First Page

442

Last Page

451

Document Type

Article

Publication Date

3-1-2014

Department

Pharmacology

Keywords

Animals, Antibodies, Neutralizing, Cell Adhesion, Cell Hypoxia, Cell Proliferation, Cytochrome P-450 CYP4A, Cytochrome P-450 Enzyme System, Endothelium, Vascular, Feedback, Physiological, Female, Fibronectins, Humans, Hydroxyeicosatetraenoic Acids, Integrin alpha4beta1, Mice, Mice, Inbred BALB C, Microvessels, Neovascularization, Physiologic, Receptors, CXCR4, Stem Cells, Vascular Endothelial Growth Factor A

Disciplines

Medicine and Health Sciences

Abstract

Circulating endothelial progenitor cells (EPC) contribute to postnatal neovascularization. We identified the cytochrome P450 4A/F-20-hydroxyeicosatetraenoic acid (CYP4A/F-20-HETE) system as a novel regulator of EPC functions associated with angiogenesis in vitro. Here, we explored cellular mechanisms by which 20-HETE regulates EPC angiogenic functions and assessed its contribution to EPC-mediated angiogenesis in vivo. Results showed that both hypoxia and vascular endothelial growth factor (VEGF) induce CYP4A11 gene and protein expression (the predominant 20-HETE synthases in human EPC), and this is accompanied by an increase in 20-HETE production by ~1.4- and 1.8-fold, respectively, compared with the control levels. Additional studies demonstrated that 20-HETE and VEGF have a synergistic effect on EPC proliferation, whereas 20-HETE antagonist 20-HEDGE or VEGF-neutralizing antibody negated 20-HETE- or VEGF-induced proliferation, respectively. These findings are consistent with the presence of a positive feedback regulation on EPC proliferation between the 20-HETE and the VEGF pathways. Furthermore, we found that 20-HETE induced EPC adhesion to fibronectin and endothelial cell monolayer by 40 ± 5.6 and 67 ± 10%, respectively, which was accompanied by a rapid induction of very late antigen-4 and chemokine receptor type 4 mRNA and protein expression. Basal and 20-HETE-stimulated increases in adhesion were negated by the inhibition of the CYP4A-20-HETE system. Lastly, EPC increased angiogenesis in vivo by 3.6 ± 0.2-fold using the Matrigel plug angiogenesis assay, and these increases were markedly reduced by the local inhibition of 20-HETE system. These results strengthened the notion that 20-HETE regulates the angiogenic functions of EPC in vitro and EPC-mediated angiogenesis in vivo.

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