Smooth Muscle-Specific Expression of CYP4A1 Induces Endothelial Sprouting in Renal Arterial Microvessels

Miao Jiang, Alexandre Mezentsev, Rowena Kemp, Kihwan Byun, John R. Falck, Joseph M. Miano, Alberto Nasjletti, Nader G. Abraham, Michal Laniado-Schwartzman

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

Cytochrome P450 (CYP) 4A1 has been characterized as the most efficient arachidonic acid ω-hydroxylase catalyzing the formation of 20-hydroxyeicosatetraenoic acid (20-HETE), a potent constrictor of the renal and cerebral microcirculation and a mitogen for smooth muscle cells. We constructed adenoviruses expressing the CYP4A1 cDNA or LacZ under the control of the smooth muscle cell-specific promoter SM22α (Ad-SM22-4A1 and Ad-SM22-nLacZ, respectively). β-Galactosidase expression was detected in Ad-SM22-nLacZ-transduced vascular smooth muscle A7r5 and PAC1 cells, but not in Ad-SM22-nLacZ-transduced 3T3 fibroblasts or vascular endothelial cells. Likewise, CYP4A1 mRNA and protein were detected in Ad-SM22-4A1-transduced A7r5 and PAC1 cells. Ad-SM22-4A1-transduced A7r5 cells metabolized lauric acid to 12-hydroxy-lauric acid at a rate 5 times greater than that of cells transduced with Ad-SM22-nLacZ (4.79±1.77 versus 0.97±0.57 nmol 12-hydroxy lauric acid/106 cells per h). Smooth muscle-specific LacZ expression was also detected in microdissected renal interlobar arteries transduced with Ad-SM22-nLacZ. Arteries transduced with Ad-SM22-4A1 produced higher levels of 20-HETE (4.04±0.29 and 13.43±2.84 ng/mg protein in Ad-SM22-nLacZ-transduced and Ad-SM22-4A1-transduced arteries, respectively) and demonstrated a marked angiogenic activity measured as the total length of sprouting neovessels (12.63±3.66 mm in Ad-SM22-4A1-transduced vessels versus 1.79±0.89 mm in Ad-SM22-nLacZ-transduced vessels). This angiogenic activity represented endothelial cell sprouting and was fully blocked by treatment with HET0016, a selective inhibitor of CYP4A-catalyzed reactions. The inhibitory effect of HET0016 was reversed by addition of a 20-HETE agonist. We conclude that Ad-SM22-4A1 drives a smooth muscle-specific functional expression of CYP4A1 and demonstrates increased angiogenesis, presumably via increased production of 20-HETE.

Original languageEnglish (US)
Pages (from-to)167-174
Number of pages8
JournalCirculation research
Volume94
Issue number2
DOIs
StatePublished - Feb 6 2004

Keywords

  • 20-HETE
  • Adenovirus
  • Angiogenesis
  • Cytochrome P450

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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    Jiang, M., Mezentsev, A., Kemp, R., Byun, K., Falck, J. R., Miano, J. M., Nasjletti, A., Abraham, N. G., & Laniado-Schwartzman, M. (2004). Smooth Muscle-Specific Expression of CYP4A1 Induces Endothelial Sprouting in Renal Arterial Microvessels. Circulation research, 94(2), 167-174. https://doi.org/10.1161/01.RES.0000111523.12842.FC