Mesenchymal Stem Cells Deliver Exogenous MicroRNA-let7c via Exosomes to Attenuate Renal Fibrosis

Mol Ther. 2016 Aug;24(7):1290-301. doi: 10.1038/mt.2016.90. Epub 2016 May 18.

Abstract

The advancement of microRNA (miRNA) therapies has been hampered by difficulties in delivering miRNA to the injured kidney in a robust and sustainable manner. Using bioluminescence imaging in mice with unilateral ureteral obstruction (UUO), we report that mesenchymal stem cells (MSCs), engineered to overexpress miRNA-let7c (miR-let7c-MSCs), selectively homed to damaged kidneys and upregulated miR-let7c gene expression, compared with nontargeting control (NTC)-MSCs. miR-let7c-MSC therapy attenuated kidney injury and significantly downregulated collagen IVα1, metalloproteinase-9, transforming growth factor (TGF)-β1, and TGF-β type 1 receptor (TGF-βR1) in UUO kidneys, compared with controls. In vitro analysis confirmed that the transfer of miR-let7c from miR-let7c-MSCs occurred via secreted exosomal uptake, visualized in NRK52E cells using cyc3-labeled pre-miRNA-transfected MSCs with/without the exosomal inhibitor, GW4869. The upregulated expression of fibrotic genes in NRK52E cells induced by TGF-β1 was repressed following the addition of isolated exosomes or indirect coculture of miR-let7c-MSCs, compared with NTC-MSCs. Furthermore, the cotransfection of NRK52E cells using the 3'UTR of TGF-βR1 confirmed that miR-let7c attenuates TGF-β1-driven TGF-βR1 gene expression. Taken together, the effective antifibrotic function of engineered MSCs is able to selectively transfer miR-let7c to damaged kidney cells and will pave the way for the use of MSCs for therapeutic delivery of miRNA targeted at kidney disease.

MeSH terms

  • Actins / metabolism
  • Animals
  • Biological Transport
  • Cell Engineering
  • Collagen / metabolism
  • Disease Models, Animal
  • ErbB Receptors / metabolism
  • Exosomes / metabolism*
  • Extracellular Vesicles / metabolism
  • Fibrosis
  • Gene Expression
  • Gene Expression Regulation
  • Gene Transfer Techniques
  • Humans
  • Kidney Diseases / genetics*
  • Kidney Diseases / metabolism
  • Kidney Diseases / pathology*
  • Kidney Diseases / therapy
  • Male
  • Mesenchymal Stem Cells / metabolism*
  • Mice
  • MicroRNAs / genetics*
  • Rats
  • Transduction, Genetic

Substances

  • Actins
  • MicroRNAs
  • Collagen
  • ErbB Receptors