HIF-prolyl hydroxylase 2 inhibition enhances the efficiency of mesenchymal stem cell-based therapies for the treatment of critical limb ischemia

Stem Cells. 2014 Jan;32(1):231-43. doi: 10.1002/stem.1540.


Upregulation of hypoxia-inducible transcription factor-1α (HIF-1α), through prolyl-hydroxylase domain protein (PHD) inhibition, can be thought of as a master switch that coordinates the expression of a wide repertoire of genes involved in regulating vascular growth and remodeling. We aimed to unravel the effect of specific PHD2 isoform silencing in cell-based strategies designed to promote therapeutic revascularization in patients with critical limb ischemia (CLI). PHD2 mRNA levels were upregulated whereas that of HIF-1α were downregulated in blood cells from patients with CLI. We therefore assessed the putative beneficial effects of PHD2 silencing on human bone marrow-derived mesenchymal stem cells (hBM-MSC)-based therapy. PHD2 silencing enhanced hBM-MSC therapeutic effect in an experimental model of CLI in Nude mice, through an upregulation of HIF-1α and its target gene, VEGF-A. In addition, PHD2-transfected hBM-MSC displayed higher protection against apoptosis in vitro and increased rate of survival in the ischemic tissue, as assessed by Fluorescence Molecular Tomography. Cotransfection with HIF-1α or VEGF-A short interfering RNAs fully abrogated the beneficial effect of PHD2 silencing on the proangiogenic capacity of hBM-MSC. We finally investigated the effect of PHD2 inhibition on the revascularization potential of ischemic targeted tissues in the diabetic pathological context. Inhibition of PHD-2 with shRNAs increased postischemic neovascularization in diabetic mice with CLI. This increase was associated with an upregulation of proangiogenic and proarteriogenic factors and was blunted by concomitant silencing of HIF-1α. In conclusion, silencing of PHD2, by the transient upregulation of HIF-1α and its target gene VEGF-A, might improve the efficiency of hBM-MSC-based therapies.

Trial registration: ClinicalTrials.gov NCT00377897 NCT01038700.

Keywords: Angiogenesis; Growth factor; Hypoxia; Ischemia; Mesenchymal stem cells; Transcription factor.

Publication types

  • Clinical Trial, Phase I
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • Apoptosis / physiology
  • Case-Control Studies
  • Cell Transplantation / methods*
  • Disease Models, Animal
  • Endovascular Procedures / methods
  • Hindlimb / blood supply*
  • Humans
  • Hypoxia-Inducible Factor-Proline Dioxygenases / antagonists & inhibitors*
  • Ischemia / enzymology
  • Ischemia / therapy*
  • Limb Salvage / methods
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Nude
  • Middle Aged
  • Prolyl-Hydroxylase Inhibitors / therapeutic use*
  • Transfection


  • Prolyl-Hydroxylase Inhibitors
  • EGLN1 protein, human
  • Hypoxia-Inducible Factor-Proline Dioxygenases

Associated data

  • ClinicalTrials.gov/NCT00377897
  • ClinicalTrials.gov/NCT01038700