title: 20-HETE regulates the angiogenic functions of human endothelial progenitor cells and contributes to angiogenesis in vivo.
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.
authors.affiliation: Department of Pharmacology, School of Medicine, Wuhan University, Wuhan, PR China (L.C., J.Y., A.M.G.); Department of Pharmacology, New York Medical College, Valhalla, New York (L.C., R.A.,M.S., K.H.G., M.L.S., A.M.G.); Department of Obstetrics and Gynecology, Westchester Medical Center, Valhalla, New York (M.K.); Obstetrics and Gynecology, Phelps Memorial Hospital Center, Sleepy Hollow, New York (L.G.M.); University of Texas Southwestern Medical Center, Dallas, Texas (J.R.F.); and Henry Ford Hospital, Detroit, Michigan (A.S.A., A.G.S.).
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.forename: Rachel
authors.initials: R
authors.lastname: Ackerman
authors.forename: Mohamed
authors.initials: M
authors.lastname: Saleh
authors.forename: Katherine H
authors.initials: KH
authors.lastname: Gotlinger
authors.forename: Michael
authors.initials: M
authors.lastname: Kessler
authors.forename: Lawrence G
authors.initials: LG
authors.lastname: Mendelowitz
authors.forename: John R
authors.initials: JR
authors.lastname: Falck
authors.forename: Ali S
authors.initials: AS
authors.lastname: Arbab
authors.forename: A Guillermo
authors.initials: AG
authors.lastname: Scicli
authors.forename: Michal L
authors.initials: ML
authors.lastname: Schwartzman
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Austin M
authors.initials: AM
authors.lastname: Guo
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000494:pharmacology; D002448:Cell Adhesion; D015687:Cell Hypoxia; D049109:Cell Proliferation; D042926:Cytochrome P-450 CYP4A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D025461:Feedback, Physiological; D005260:Female; D005353:Fibronectins / Q000502:physiology; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000037:antagonists & inhibitors / Q000378:metabolism*; D039041:Integrin alpha4beta1 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D055806:Microvessels / Q000502:physiology; D018919:Neovascularization, Physiologic; D019718:Receptors, CXCR4 / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology
chemical_list: D057134:Antibodies, Neutralizing; D005353:Fibronectins; D006893:Hydroxyeicosatetraenoic Acids; D039041:Integrin alpha4beta1; D019718:Receptors, CXCR4; D042461:Vascular Endothelial Growth Factor A; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D003577:Cytochrome P-450 Enzyme System; C465478:CYP4A11 protein, human; D042926:Cytochrome P-450 CYP4A