Pravastatin reverses obesity-induced dysfunction of induced pluripotent stem cell-derived endothelial cells via a nitric oxide-dependent mechanism

Eur Heart J. 2015 Apr 1;36(13):806-16. doi: 10.1093/eurheartj/ehu411. Epub 2014 Nov 2.

Abstract

Aims: High-fat diet-induced obesity (DIO) is a major contributor to type II diabetes and micro- and macro-vascular complications leading to peripheral vascular disease (PVD). Metabolic abnormalities of induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) from obese individuals could potentially limit their therapeutic efficacy for PVD. The aim of this study was to compare the function of iPSC-ECs from normal and DIO mice using comprehensive in vitro and in vivo assays.

Methods and results: Six-week-old C57Bl/6 mice were fed with a normal or high-fat diet. At 24 weeks, iPSCs were generated from tail tip fibroblasts and differentiated into iPSC-ECs using a directed monolayer approach. In vitro functional analysis revealed that iPSC-ECs from DIO mice had significantly decreased capacity to form capillary-like networks, diminished migration, and lower proliferation. Microarray and ELISA confirmed elevated apoptotic, inflammatory, and oxidative stress pathways in DIO iPSC-ECs. Following hindlimb ischaemia, mice receiving intramuscular injections of DIO iPSC-ECs had significantly decreased reperfusion compared with mice injected with control healthy iPSC-ECs. Hindlimb sections revealed increased muscle atrophy and presence of inflammatory cells in mice receiving DIO iPSC-ECs. When pravastatin was co-administered to mice receiving DIO iPSC-ECs, a significant increase in reperfusion was observed; however, this beneficial effect was blunted by co-administration of the nitric oxide synthase inhibitor, N(ω)-nitro-l-arginine methyl ester.

Conclusion: This is the first study to provide evidence that iPSC-ECs from DIO mice exhibit signs of endothelial dysfunction and have suboptimal efficacy following transplantation in a hindlimb ischaemia model. These findings may have important implications for future treatment of PVD using iPSC-ECs in the obese population.

Keywords: Diet-induced obesity; Endothelial cells; Hindlimb ischaemia; Induced pluripotent stem cells; Peripheral vascular disease; Statins.

Publication types

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

MeSH terms

  • Analysis of Variance
  • Animals
  • Apoptosis / physiology
  • Cell Differentiation
  • Diet, High-Fat
  • Endothelial Cells / physiology*
  • Enzyme Inhibitors / pharmacology
  • Fibroblasts / physiology
  • Hindlimb / blood supply
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacology*
  • Induced Pluripotent Stem Cells / physiology*
  • Injections, Intramuscular
  • Ischemia / physiopathology
  • Ischemia / prevention & control
  • Mice, Inbred C57BL
  • Muscle, Skeletal
  • Muscular Diseases / prevention & control
  • NG-Nitroarginine Methyl Ester / pharmacology
  • Nitric Oxide / physiology*
  • Nitric Oxide Synthase Type III / metabolism
  • Obesity / physiopathology*
  • Oxidative Stress / physiology
  • Pravastatin / pharmacology*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Random Allocation
  • Reperfusion Injury / physiopathology
  • Signal Transduction

Substances

  • Enzyme Inhibitors
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • Nitric Oxide
  • Nitric Oxide Synthase Type III
  • Proto-Oncogene Proteins c-akt
  • Pravastatin
  • NG-Nitroarginine Methyl Ester