Interferon-gamma induces prolyl hydroxylase (PHD)3 through a STAT1-dependent mechanism in human endothelial cells

Arterioscler Thromb Vasc Biol. 2009 Sep;29(9):1363-9. doi: 10.1161/ATVBAHA.109.192542. Epub 2009 Jul 2.

Abstract

Objective: We previously reported that interferons (IFNs) regulate transcription of HIF-1alpha in human endothelial cells (ECs), linking immunity and hypoxia. Prolyl hydroxylases (PHDs) regulate expression of HIF-1alpha in response to hypoxia. We examined whether IFNs affect PHD expression and whether PHDs regulate the EC response to IFNs.

Methods and results: Human cell cultures were treated with various cytokines, and PHD expression was examined using qRT-PCR and immunoblotting. IFNgamma and, to a lesser extent, IFNalpha significantly induced PHD3, but not PHD1 or 2, mRNA, and protein expression selectively in ECs directly via a JAK/STAT1 pathway as demonstrated by pharmacological inhibition, siRNA knockdown, and chromatin immunoprecipitation. Inhibition of PHD activity with dimethyloxallyl glycine or desferroxamine reduced IFNg-dependent responses in these same cells.

Conclusions: IFNgamma induces PHD3 through a JAK/STAT1-dependent mechanism in human ECs. Induction is independent of HIF-1alpha and may contribute to expression of IFNgamma-dependent genes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids, Dicarboxylic / pharmacology
  • Cell Hypoxia
  • Cells, Cultured
  • Deferoxamine / pharmacology
  • Dioxygenases / antagonists & inhibitors
  • Dioxygenases / biosynthesis*
  • Dioxygenases / genetics
  • Endothelial Cells / drug effects
  • Endothelial Cells / enzymology*
  • Enzyme Induction
  • Enzyme Inhibitors / pharmacology
  • Genes, Immediate-Early
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Hypoxia-Inducible Factor-Proline Dioxygenases
  • Interferon-alpha / metabolism
  • Interferon-gamma / metabolism*
  • Janus Kinases / metabolism
  • RNA Interference
  • RNA, Messenger / biosynthesis
  • STAT1 Transcription Factor / genetics
  • STAT1 Transcription Factor / metabolism*
  • Signal Transduction
  • Time Factors
  • Transcription, Genetic

Substances

  • Amino Acids, Dicarboxylic
  • Enzyme Inhibitors
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Interferon-alpha
  • RNA, Messenger
  • STAT1 Transcription Factor
  • STAT1 protein, human
  • Interferon-gamma
  • Dioxygenases
  • EGLN3 protein, human
  • Hypoxia-Inducible Factor-Proline Dioxygenases
  • Janus Kinases
  • Deferoxamine
  • oxalylglycine