miR-200a Prevents Renal Fibrogenesis Through Repression of TGF-β2 Expression
- Bo Wang1,
- Philip Koh1,
- Catherine Winbanks2,
- Melinda T. Coughlan1,
- Aaron McClelland1,
- Anna Watson1,
- Karin Jandeleit-Dahm1,
- Wendy C. Burns1,
- Merlin C. Thomas1,
- Mark E. Cooper1 and
- Phillip Kantharidis1
- 1Diabetes Division, JDRF Danielle Alberti Memorial Centre for Diabetes Complications, Melbourne, Australia;
- 2Muscle Biology and Therapeutics, Baker IDI Heart and Diabetes Institute, Melbourne, Australia.
- Corresponding author: Phillip Kantharidis, .
OBJECTIVE Progressive fibrosis in the diabetic kidney is driven and sustained by a diverse range of profibrotic factors. This study examines the critical role of microRNAs (miRNAs) in the regulation of the key fibrotic mediators, TGF-β1 and TGF-β2.
RESEARCH DESIGN AND METHODS Rat proximal-tubular epithelial cells (NRK52E) were treated with TGF-β1 and TGF-β2 for 3 days, and expression of markers of epithelial-to-mesenchymal transition (EMT) and fibrogenesis were assessed by RT-PCR and Western blotting. The expression of miR-141 and miR-200a was also assessed, as was their role as translational repressors of TGF-β signaling. Finally, these pathways were explored in two different mouse models, representing early and advanced diabetic nephropathy.
RESULTS Both TGF-β1 and TGF-β2 induced EMT and fibrogenesis in NRK52E cells. TGF-β1 and TGF-β2 also downregulated expression of miR-200a. The importance of these changes was demonstrated by the finding that ectopic expression miR-200a downregulated smad-3 activity and the expression of matrix proteins and prevented TGF-β–dependent EMT. miR-200a also downregulated the expression of TGF-β2, via direct interaction with the 3′ untranslated region of TGF-β2. The renal expression of miR-141 and miR-200a was also reduced in mouse models representing early and advanced kidney disease.
CONCLUSIONS miR-200a and miR-141 significantly impact on the development and progression of TGF-β–dependent EMT and fibrosis in vitro and in vivo. These miRNAs appear to be intricately involved in fibrogenesis, both as downstream mediators of TGF-β signaling and as components of feedback regulation, and as such represent important new targets for the prevention of progressive kidney disease in the context of diabetes.
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- Received June 28, 2010.
- Accepted October 1, 2010.
- © 2011 by the American Diabetes Association.
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