Downregulation of endothelial microRNA-200b supports cutaneous wound angiogenesis by desilencing GATA binding protein 2 and vascular endothelial growth factor receptor 2

Arterioscler Thromb Vasc Biol. 2012 Jun;32(6):1372-82. doi: 10.1161/ATVBAHA.112.248583. Epub 2012 Apr 12.

Abstract

Objective: MicroRNAs (miRs) regulate angiogenesis by posttranscriptional silencing of target genes. The significance of angiostatic miR-200b in switching on skin wound angiogenesis was tested.

Methods and results: Wounding caused imminent and transient downregulation of miR-200b in dermal wound-edge endothelial cells. Derailing this injury response by lentiviral delivery of miR-200b in vivo impaired wound angiogenesis. Computational prediction, target reporter luciferase assay, and Western blot analysis provided first evidence that miR-200b targets globin transcription factor binding protein 2 (GATA2) and vascular endothelial growth factor receptor 2 (VEGFR2). Overexpression of GATA2 or VEGFR2 in endothelial cells rescued the angiostatic effect of miR-200b in vitro. Downregulation of miR-200b derepressed GATA2 and VEGFR2 expression to switch on wound angiogenesis, which was disrupted in diabetic wounds. Treatment of endothelial cells with tumor necrosis factor-α, a proinflammatory cytokine abundant in diabetic wounds, induced miR-200b expression, silenced GATA2 and VEGFR2, and suppressed angiogenesis. These outcomes were attenuated using anti-miR-200b strategy. Neutralization of tumor necrosis factor-α in the diabetic wounds improved wound angiogenesis and closure, which was accompanied by downregulation of miR-200b expression and desilencing of GATA2 and VEGFR2.

Conclusions: Injury-induced repression of miR-200b turned on wound angiogenesis. In mice with diabetes mellitus,excessive tumor necrosis factor-α induced miR-200b blunting proangiogenic functions of GATA2 and VEGFR2.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Blotting, Western
  • Diabetes Mellitus / genetics
  • Diabetes Mellitus / metabolism
  • Diabetes Mellitus / pathology
  • Diabetes Mellitus / therapy*
  • Disease Models, Animal
  • Down-Regulation
  • Endothelial Cells / metabolism
  • GATA2 Transcription Factor / genetics
  • GATA2 Transcription Factor / metabolism*
  • Genes, Reporter
  • Genetic Therapy*
  • Genetic Vectors
  • HEK293 Cells
  • Humans
  • Injections, Intradermal
  • Laser-Doppler Flowmetry
  • Lentivirus / genetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / metabolism*
  • Mutation
  • Neovascularization, Physiologic*
  • RNA Interference
  • RNA, Messenger / metabolism
  • Receptors, Leptin / genetics
  • Receptors, Leptin / metabolism
  • Receptors, Tumor Necrosis Factor, Type I / administration & dosage
  • Regional Blood Flow
  • Signal Transduction
  • Skin / blood supply
  • Skin / injuries
  • Skin / metabolism*
  • Skin / pathology
  • Time Factors
  • Transfection
  • Tumor Necrosis Factor-alpha / metabolism
  • Vascular Endothelial Growth Factor Receptor-2 / genetics
  • Vascular Endothelial Growth Factor Receptor-2 / metabolism*
  • Wound Healing*
  • Wounds, Penetrating / genetics
  • Wounds, Penetrating / metabolism
  • Wounds, Penetrating / pathology
  • Wounds, Penetrating / therapy*

Substances

  • GATA2 Transcription Factor
  • GATA2 protein, human
  • Gata2 protein, mouse
  • MIRN200 microRNA, human
  • MicroRNAs
  • Mirn200 microRNA, mouse
  • RNA, Messenger
  • Receptors, Leptin
  • Receptors, Tumor Necrosis Factor, Type I
  • Tumor Necrosis Factor-alpha
  • Vascular Endothelial Growth Factor Receptor-2