title: Pravastatin reverses obesity-induced dysfunction of induced pluripotent stem cell-derived endothelial cells via a nitric oxide-dependent mechanism.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gu
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Mordwinkin
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA Department of Vascular Surgery, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Nigel G
authors.initials: NG
authors.lastname: Kooreman
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Jaecheol
authors.initials: J
authors.lastname: Lee
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Haodi
authors.initials: H
authors.lastname: Wu
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Shijun
authors.initials: S
authors.lastname: Hu
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Jared M
authors.initials: JM
authors.lastname: Churko
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Diecke
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Burridge
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Chunjiang
authors.initials: C
authors.lastname: He
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Frances E
authors.initials: FE
authors.lastname: Barron
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Sang-Ging
authors.initials: SG
authors.lastname: Ong
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Gold
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA joewu@stanford.edu.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002454:Cell Differentiation; D059305:Diet, High-Fat; D042783:Endothelial Cells / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology; D006614:Hindlimb / Q000098:blood supply; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D007273:Injections, Intramuscular; D007511:Ischemia / Q000503:physiopathology / Q000517:prevention & control; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal; D009135:Muscular Diseases / Q000517:prevention & control; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009765:Obesity / Q000503:physiopathology*; D018384:Oxidative Stress / Q000502:physiology; D017035:Pravastatin / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011897:Random Allocation; D015427:Reperfusion Injury / Q000503:physiopathology; D015398:Signal Transduction
chemical_list: D004791:Enzyme Inhibitors; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D051057:Proto-Oncogene Proteins c-akt; D017035:Pravastatin; D019331:NG-Nitroarginine Methyl Ester
keywords: Diet-induced obesity; Endothelial cells; Hindlimb ischaemia; Induced pluripotent stem cells; Peripheral vascular disease; Statins