Vascular characterization of mice with endothelial expression of cytochrome P450 4F2

Jennifer Cheng, Matthew L. Edin, Samantha L. Hoopes, Hong Li, J. Alyce Bradbury, Joan P. Graves, Laura M. DeGraff, Fred B. Lih, Victor Garcia, Jafar Sadik B Shaik, Kenneth B. Tomer, Gordon P. Flake, John R. Falck, Craig R. Lee, Samuel M. Poloyac, Michal L. Schwartzman, Darryl C. Zeldin

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Cytochrome P450 (CYP) 4A and 4F enzymes metabolize arachidonic acid to 20-hydroxyeico-satetraenoic acid (20-HETE). Although CYP4A-derived 20-HETE is known to have prohypertensive and proan-giogenic properties, the effects of CYP4F-derived metabolites are not well characterized. To investigate the role of CYP4F2 in vascular disease, we generated mice with endothelial expression of human CYP4F2 (Tie2-CYP4F2-Tr). LC/MS/MS analysis revealed 2-fold increases in 20-HETE levels in tissues and endothelial cells (ECs), relative to wild-type (WT) controls. Tie2-CYP4F2-Tr ECs demonstrated increases in growth (267.1 ±33.4 vs. 205.0±13% at 48 h) and tube formation (7.7±1.1 vs. 1.6±0.5 tubes/field) that were 20-HETE dependent and associated with up-regulation of prooxidant NADPH oxidase and proangiogenic VEGF. Increases in VEGF and NADPH oxidase levels were abrogated by inhibitors of NADPH oxidase and MAPK, respectively, suggesting the possibility of crosstalk between pathways. Interestingly, IL-6 levels in Tie2-CYP4F2-Tr mice (18.6±2.7 vs. 7.9±2.7 pg/ml) were up-regulated via NADPH oxidase- and 20-HETE-depen-dent mechanisms. Although Tie2-CYP4F2-Tr aortas displayed increased vasoconstriction, vasorelaxation and blood pressure were unchanged. Our findings indicate that human CYP4F2 significantly increases 20-HETE production, CYP4F2-derived 20-HETE mediates EC proliferation and angiogenesis via VEGF- and NADPH oxidase-dependent manners, and the Tie2-CYP4F2-Tr mouse is a novel model for examining the pathophysiological effects of CYP4F2-derived 20-HETE in the vasculature.

Original languageEnglish (US)
Pages (from-to)2915-2931
Number of pages17
JournalFASEB Journal
Volume28
Issue number7
DOIs
StatePublished - Jul 1 2014

Fingerprint

Cytochrome P-450 Enzyme System
Blood Vessels
NADPH Oxidase
Acids
Endothelial cells
Vascular Endothelial Growth Factor A
Endothelial Cells
Cytochrome P-450 CYP4A
Blood pressure
Cell proliferation
Metabolites
Crosstalk
Vasoconstriction
Vascular Diseases
Arachidonic Acid
Vasodilation
Aorta
Interleukin-6
Up-Regulation
Cell Proliferation

Keywords

  • 20-HETE
  • Angiogenesis
  • Cellular proliferation
  • CYP4F2
  • Oxidative stress

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Cheng, J., Edin, M. L., Hoopes, S. L., Li, H., Bradbury, J. A., Graves, J. P., ... Zeldin, D. C. (2014). Vascular characterization of mice with endothelial expression of cytochrome P450 4F2. FASEB Journal, 28(7), 2915-2931. https://doi.org/10.1096/fj.13-241927

Vascular characterization of mice with endothelial expression of cytochrome P450 4F2. / Cheng, Jennifer; Edin, Matthew L.; Hoopes, Samantha L.; Li, Hong; Bradbury, J. Alyce; Graves, Joan P.; DeGraff, Laura M.; Lih, Fred B.; Garcia, Victor; Shaik, Jafar Sadik B; Tomer, Kenneth B.; Flake, Gordon P.; Falck, John R.; Lee, Craig R.; Poloyac, Samuel M.; Schwartzman, Michal L.; Zeldin, Darryl C.

In: FASEB Journal, Vol. 28, No. 7, 01.07.2014, p. 2915-2931.

Research output: Contribution to journalArticle

Cheng, J, Edin, ML, Hoopes, SL, Li, H, Bradbury, JA, Graves, JP, DeGraff, LM, Lih, FB, Garcia, V, Shaik, JSB, Tomer, KB, Flake, GP, Falck, JR, Lee, CR, Poloyac, SM, Schwartzman, ML & Zeldin, DC 2014, 'Vascular characterization of mice with endothelial expression of cytochrome P450 4F2', FASEB Journal, vol. 28, no. 7, pp. 2915-2931. https://doi.org/10.1096/fj.13-241927
Cheng J, Edin ML, Hoopes SL, Li H, Bradbury JA, Graves JP et al. Vascular characterization of mice with endothelial expression of cytochrome P450 4F2. FASEB Journal. 2014 Jul 1;28(7):2915-2931. https://doi.org/10.1096/fj.13-241927
Cheng, Jennifer ; Edin, Matthew L. ; Hoopes, Samantha L. ; Li, Hong ; Bradbury, J. Alyce ; Graves, Joan P. ; DeGraff, Laura M. ; Lih, Fred B. ; Garcia, Victor ; Shaik, Jafar Sadik B ; Tomer, Kenneth B. ; Flake, Gordon P. ; Falck, John R. ; Lee, Craig R. ; Poloyac, Samuel M. ; Schwartzman, Michal L. ; Zeldin, Darryl C. / Vascular characterization of mice with endothelial expression of cytochrome P450 4F2. In: FASEB Journal. 2014 ; Vol. 28, No. 7. pp. 2915-2931.
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AU - Lih, Fred B.

AU - Garcia, Victor

AU - Shaik, Jafar Sadik B

AU - Tomer, Kenneth B.

AU - Flake, Gordon P.

AU - Falck, John R.

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N2 - Cytochrome P450 (CYP) 4A and 4F enzymes metabolize arachidonic acid to 20-hydroxyeico-satetraenoic acid (20-HETE). Although CYP4A-derived 20-HETE is known to have prohypertensive and proan-giogenic properties, the effects of CYP4F-derived metabolites are not well characterized. To investigate the role of CYP4F2 in vascular disease, we generated mice with endothelial expression of human CYP4F2 (Tie2-CYP4F2-Tr). LC/MS/MS analysis revealed 2-fold increases in 20-HETE levels in tissues and endothelial cells (ECs), relative to wild-type (WT) controls. Tie2-CYP4F2-Tr ECs demonstrated increases in growth (267.1 ±33.4 vs. 205.0±13% at 48 h) and tube formation (7.7±1.1 vs. 1.6±0.5 tubes/field) that were 20-HETE dependent and associated with up-regulation of prooxidant NADPH oxidase and proangiogenic VEGF. Increases in VEGF and NADPH oxidase levels were abrogated by inhibitors of NADPH oxidase and MAPK, respectively, suggesting the possibility of crosstalk between pathways. Interestingly, IL-6 levels in Tie2-CYP4F2-Tr mice (18.6±2.7 vs. 7.9±2.7 pg/ml) were up-regulated via NADPH oxidase- and 20-HETE-depen-dent mechanisms. Although Tie2-CYP4F2-Tr aortas displayed increased vasoconstriction, vasorelaxation and blood pressure were unchanged. Our findings indicate that human CYP4F2 significantly increases 20-HETE production, CYP4F2-derived 20-HETE mediates EC proliferation and angiogenesis via VEGF- and NADPH oxidase-dependent manners, and the Tie2-CYP4F2-Tr mouse is a novel model for examining the pathophysiological effects of CYP4F2-derived 20-HETE in the vasculature.

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