MicroRNAs (miRNAs) are regulators of myriad cellular events, but evidence for a single miRNA that can efficiently differentiate multipotent stem cells into a specific lineage or regulate direct reprogramming of cells into an alternative cell fate has been elusive. Here we show that miR-145 and miR-143 are co-transcribed in multipotent murine cardiac progenitors before becoming localized to smooth muscle cells, including neural crest stem-cell-derived vascular smooth muscle cells. miR-145 and miR-143 were direct transcriptional targets of serum response factor, myocardin and Nkx2-5 (NK2 transcription factor related, locus 5) and were downregulated in injured or atherosclerotic vessels containing proliferating, less differentiated smooth muscle cells. miR-145 was necessary for myocardin-induced reprogramming of adult fibroblasts into smooth muscle cells and sufficient to induce differentiation of multipotent neural crest stem cells into vascular smooth muscle. Furthermore, miR-145 and miR-143 cooperatively targeted a network of transcription factors, including Klf4 (Kruppel-like factor 4), myocardin and Elk-1 (ELK1, member of ETS oncogene family), to promote differentiation and repress proliferation of smooth muscle cells. These findings demonstrate that miR-145 can direct the smooth muscle fate and that miR-145 and miR-143 function to regulate the quiescent versus proliferative phenotype of smooth muscle cells.
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We thank R. Blelloch for DGCR8-null EBs; R.J. Schwartz for SRF-null ES cells; I. Charo and N. Saederup for RNA from atherosclerotic tissue; J. Maurer for JoMa neural crest cell line; L. Qian and Y. Huang for providing mouse cardiac infarct RNA; C. Tsou for help with calcium flux assays; E. N. Olson for the myocardin expression plasmid; P. Swinton for generation of transgenic mice; J. Fish and C. Miller for histopathology support; S. Ordway and G. Howard for scientific editing; B. Taylor for manuscript preparation. We also thank members of the Srivastava laboratory for discussions. J.M.M. was supported by HL62572 and HL091168 from NHLBI/NIH. D.S. was supported by grants from the NHLBI/NIH and the California Institute for Regenerative Medicine (CIRM) and was an Established Investigator of the American Heart Association. This work was also supported by NIH/NCRR grant C06 RR018928 to the Gladstone Institutes.
Author Contributions K.R.C. and D.S. designed the study and K.R.C. executed or oversaw execution of all experiments; N.T.S. and E.C.B. performed the NCC studies; M.P.W. and K.N.I. performed some expression and stem cell studies and K.N.I. helped supervise the project; A.N.M. provided technical support; T.-H.L. and J.M.M. performed carotid artery ligation studies; S.U.M. isolated YFP+ progenitor cells and performed some expression studies; J.M.M. assisted K.R.C and D.S. in editing the manuscript; K.R.C. and D.S. wrote the manuscript and D.S. supervised all aspects of the project.
D.S. serves on the Scientific Advisory Board of iZumi Bio.
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Cordes, K., Sheehy, N., White, M. et al. miR-145 and miR-143 regulate smooth muscle cell fate and plasticity. Nature 460, 705–710 (2009). https://doi.org/10.1038/nature08195
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