MicroRNA miR-27b rescues bone marrow-derived angiogenic cell function and accelerates wound healing in type 2 diabetes mellitus

Arterioscler Thromb Vasc Biol. 2014 Jan;34(1):99-109. doi: 10.1161/ATVBAHA.113.302104. Epub 2013 Oct 31.

Abstract

Objective: Vascular precursor cells with angiogenic potentials are important for tissue repair, which is impaired in diabetes mellitus. MicroRNAs are recently discovered key regulators of gene expression, but their role in vascular precursor cell-mediated angiogenesis in diabetes mellitus is unknown. We tested the hypothesis that the microRNA miR-27b rescues impaired bone marrow-derived angiogenic cell (BMAC) function in vitro and in vivo in type 2 diabetic mice.

Approach and results: BMACs from adult male type 2 diabetic db/db and from normal littermate db/+ mice were used. miR-27b expression was decreased in db/db BMACs. miR-27b mimic improved db/db BMAC function, including proliferation, adhesion, tube formation, and delayed apoptosis, but it did not affect migration. Elevated thrombospondin-1 (TSP-1) protein in db/db BMACs was suppressed on miR-27b mimic transfection. Inhibition of miR-27b in db/+ BMACs reduced angiogenesis, which was reversed by TSP-1 small interfering RNA (siRNA). miR-27b suppressed the pro-oxidant protein p66(shc) and mitochondrial oxidative stress, contributing to its protection of BMAC function. miR-27b also suppressed semaphorin 6A to improve BMAC function in diabetes mellitus. Luciferase binding assay suggested that miR-27b directly targeted TSP-1, TSP-2, p66(shc), and semaphorin 6A. miR-27b improved topical cell therapy of diabetic BMACs on diabetic skin wound closure, with a concomitant augmentation of wound perfusion and capillary formation. Normal BMAC therapy with miR-27b inhibition demonstrated reduced efficacy in wound closure, perfusion, and capillary formation. Local miR-27b delivery partly improved wound healing in diabetic mice.

Conclusions: miR-27b rescues impaired BMAC angiogenesis via TSP-1 suppression, semaphorin 6A expression, and p66shc-dependent mitochondrial oxidative stress and improves BMAC therapy in wound healing in type 2 diabetic mice.

Keywords: diabetes mellitus; microRNAs; type 2; wound healing.

Publication types

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

MeSH terms

  • Animals
  • Bone Marrow Cells / metabolism*
  • Bone Marrow Cells / pathology
  • CD36 Antigens / deficiency
  • CD36 Antigens / genetics
  • CD47 Antigen / genetics
  • CD47 Antigen / metabolism
  • Case-Control Studies
  • Cells, Cultured
  • Diabetes Mellitus, Type 2 / genetics
  • Diabetes Mellitus, Type 2 / metabolism*
  • Diabetes Mellitus, Type 2 / pathology
  • Diabetes Mellitus, Type 2 / physiopathology
  • Diabetes Mellitus, Type 2 / therapy
  • Disease Models, Animal
  • Female
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • MicroRNAs / antagonists & inhibitors
  • MicroRNAs / blood
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Middle Aged
  • Mitochondria / metabolism
  • Neovascularization, Physiologic*
  • Oligoribonucleotides / pharmacology
  • Oxidative Stress
  • RNA Interference
  • Semaphorins / genetics
  • Semaphorins / metabolism
  • Shc Signaling Adaptor Proteins / genetics
  • Shc Signaling Adaptor Proteins / metabolism
  • Signal Transduction
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Stem Cells / metabolism*
  • Stem Cells / pathology
  • Thrombospondin 1 / genetics
  • Thrombospondin 1 / metabolism
  • Thrombospondins / genetics
  • Thrombospondins / metabolism
  • Time Factors
  • Transfection
  • Wound Healing*

Substances

  • CD36 Antigens
  • CD47 Antigen
  • Cd47 protein, mouse
  • MIRN27 microRNA, human
  • MicroRNAs
  • Mirn27 microRNA, mouse
  • Oligoribonucleotides
  • Sema6a protein, mouse
  • Semaphorins
  • Shc Signaling Adaptor Proteins
  • Shc1 protein, mouse
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Thrombospondin 1
  • Thrombospondins
  • thrombospondin 2
  • Thbs1 protein, mouse