MicroRNAs comprise a broad class of small non-coding RNAs that control expression of complementary target messenger RNAs1,2. Dysregulation of microRNAs by several mechanisms has been described in various disease states3,4,5 including cardiac disease6,7,8,9,10. Whereas previous studies of cardiac disease have focused on microRNAs that are primarily expressed in cardiomyocytes, the role of microRNAs expressed in other cell types of the heart is unclear. Here we show that microRNA-21 (miR-21, also known as Mirn21) regulates the ERK–MAP kinase signalling pathway in cardiac fibroblasts, which has impacts on global cardiac structure and function. miR-21 levels are increased selectively in fibroblasts of the failing heart, augmenting ERK–MAP kinase activity through inhibition of sprouty homologue 1 (Spry1). This mechanism regulates fibroblast survival and growth factor secretion, apparently controlling the extent of interstitial fibrosis and cardiac hypertrophy. In vivo silencing of miR-21 by a specific antagomir in a mouse pressure-overload-induced disease model reduces cardiac ERK–MAP kinase activity, inhibits interstitial fibrosis and attenuates cardiac dysfunction. These findings reveal that microRNAs can contribute to myocardial disease by an effect in cardiac fibroblasts. Our results validate miR-21 as a disease target in heart failure and establish the therapeutic efficacy of microRNA therapeutic intervention in a cardiovascular disease setting.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Ambros, V. The functions of animal microRNAs. Nature 431, 350–355 (2004)
Bartel, D. P. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297 (2004)
Mi, S. et al. MicroRNA expression signatures accurately discriminate acute lymphoblastic leukemia from acute myeloid leukemia. Proc. Natl Acad. Sci. USA 104, 19971–19976 (2007)
He, L. et al. A microRNA component of the p53 tumour suppressor network. Nature 447, 1130–1134 (2007)
Huang, J. et al. Cellular microRNAs contribute to HIV-1 latency in resting primary CD4+ T lymphocytes. Nature Med. 13, 1241–1247 (2007)
Care, A. et al. MicroRNA-133 controls cardiac hypertrophy. Nature Med. 13, 613–618 (2007)
van Rooij, E. et al. Control of stress-dependent cardiac growth and gene expression by a microRNA. Science 316, 575–579 (2007)
Yang, B. et al. The muscle-specific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2. Nature Med. 13, 486–491 (2007)
Zhao, Y. et al. Dysregulation of cardiogenesis, cardiac conduction, and cell cycle in mice lacking miRNA-1-2. Cell 129, 303–317 (2007)
Sayed, D., Hong, C., Chen, I. Y., Lypowy, J. & Abdellatif, M. MicroRNAs play an essential role in the development of cardiac hypertrophy. Circ. Res. 100, 416–424 (2007)
Engelhardt, S., Hein, L., Wiesmann, F. & Lohse, M. J. Progressive hypertrophy and heart failure in β1-adrenergic receptor transgenic mice. Proc. Natl Acad. Sci. USA 96, 7059–7064 (1999)
Cheng, Y. H. et al. MicroRNAs are aberrantly expressed in hypertrophic heart — do they play a role in cardiac hypertrophy? Am. J. Pathol. 170, 1831–1840 (2007)
Tatsuguchi, M. et al. Expression of microRNAs is dynamically regulated during cardiomyocyte hypertrophy. J. Mol. Cell. Cardiol. 42, 1137–1141 (2007)
Sayed, D. et al. MicroRNA-21 targets Sprouty2 and promotes cellular outgrowths. Mol. Biol. Cell 19, 3272–3282 (2008)
Rockman, H. A. et al. Segregation of atrial-specific and inducible expression of an atrial natriuretic factor transgene in an in vivo murine model of cardiac hypertrophy. Proc. Natl Acad. Sci. USA 88, 8277–8281 (1991)
Kudej, R. K. et al. Effects of chronic β-adrenergic receptor stimulation in mice. J. Mol. Cell. Cardiol. 29, 2735–2746 (1997)
Dudley, D. T., Pang, L., Decker, S. J., Bridges, A. J. & Saltiel, A. R. A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc. Natl Acad. Sci. USA 92, 7686–7689 (1995)
Pages, G. et al. Mitogen-activated protein kinases p42mapk and p44mapk are required for fibroblast proliferation. Proc. Natl Acad. Sci. USA 90, 8319–8323 (1993)
Hanafusa, H., Torii, S., Yasunaga, T. & Nishida, E. Sprouty1 and Sprouty2 provide a control mechanism for the Ras/MAPK signalling pathway. Nature Cell Biol. 4, 850–858 (2002)
Casci, T., Vinos, J. & Freeman, M. Sprouty, an intracellular inhibitor of Ras signaling. Cell 96, 655–665 (1999)
Basson, M. A. et al. Sprouty1 is a critical regulator of GDNF/RET-mediated kidney induction. Dev. Cell 8, 229–239 (2005)
Krutzfeldt, J. et al. Silencing of microRNAs in vivo with ‘antagomirs’. Nature 438, 685–689 (2005)
Castoldi, M. et al. A sensitive array for microRNA expression profiling (miChip) based on locked nucleic acids (LNA). RNA 12, 913–920 (2006)
Thum, T. et al. MicroRNAs in the human heart: a clue to fetal gene reprogramming in heart failure. Circulation 116, 258–267 (2007)
Buitrago, M. et al. The transcriptional repressor Nab1 is a specific regulator of pathological cardiac hypertrophy. Nature Med. 11, 837–844 (2005)
Thum, T. & Borlak, J. Mechanistic role of cytochrome P450 monooxygenases in oxidized low-density lipoprotein-induced vascular injury: therapy through LOX-1 receptor antagonism? Circ. Res. 94, e1–e13 (2004)
Kissler, S. et al. In vivo RNA interference demonstrates a role for Nramp1 in modifying susceptibility to type 1 diabetes. Nature Genet. 38, 479–483 (2006)
Li, X., Wang, W. D. & Lufkin, T. Dicistronic LacZ and alkaline phosphatase reporter constructs permit simultaneous histological analysis of expression from multiple transgenes. Biotechniques 23, 874–878 (1997)
Lewandoski, M., Meyers, E. N. & Martin, G. R. Analysis of Fgf8 gene function in vertebrate development. Cold Spring Harb. Symp. Quant. Biol. 62, 159–168 (1997)
Merkle, S. et al. A role for caspase-1 in heart failure. Circ. Res. 100, 645–653 (2007)
We thank N. Hemmrich, U. Keller, J. Schittl, C. Dienesch, S. Thum, A. Leupold, M. Kümmel, S. Schraut, A. Lauer, S. Marquart, E. Leich and A. Horn for technical assistance. We acknowledge the contribution of V. Benes and S. Schmidt (miChip microarray Platform, EMBL), D. Fraccarollo and K. Hu (in vivo studies), S. Leierseder and X. Loyer (primary fibroblast preparation), C. Sohn-Lee (in situ hybridization experiments) and M. Manoharan, R. Braich and B. Bhat (antagomir oligonucleotides). We also thank L. Field, T. Brand and M. Gessler for discussions. This work was supported in part by grants from the IZKF (E-31 to T. Thum), the Deutsche Forschungsgemeinschaft (DFG TH903/7-1 to T. Thum and J.B.), the Rudolf Virchow Center/DFG Research Center for Experimental Biomedicine (S.E., S.K.), the Bavarian Ministry of Technology, ProCorde and Sanofi-Aventis (S.E.), and the US NIH (R01 CA78711 to G.R.M.). M.C. is supported by an Excellence Fellowship of The Medical Faculty of the University of Heidelberg, M.U.M. by a Cancer Research Net grant (BMBF (NGFN) 201GS0450), and M.B. by the Leopoldina Academy (BMBF-LPD 9901/8-141).
Author Contributions T. Thum, C.G., J.F., T.F., S.K., M.B., P.G., S.J., M.C. and S.E. performed experiments. M.A.B and J.D.L. provided the Spry/LacZ mouse line. J.T.R.P., S.H.R. and T. Tuschl contributed the in situ hybridization experiments. T. Thum, C.G., J.F., W.R., S.F., J.S., V.K., A.R., M.M., G.R.M., J.B. and S.E. analysed data. T. Thum, J.B. and S.E. designed the study. T. Thum, G.R.M., J.B. and S.E. wrote the manuscript. J.B. and S.E. contributed equally as joint senior authors to the study.
T. Thum, C.G., J.B. and S.E. have submitted a patent application on the use of microRNAs in heart disease.
About this article
Circulation Research (2019)
Nano LIFE (2019)
Impact of statins on cellular respiration and de-differentiation of myofibroblasts in human failing hearts
ESC Heart Failure (2019)
Basic Research in Cardiology (2019)
Blockade of miR-140-3p prevents functional deterioration in afterload-enhanced engineered heart tissue
Scientific Reports (2019)