MicroRNA-mediated conversion of human fibroblasts to neurons

Abstract

Neurogenic transcription factors and evolutionarily conserved signalling pathways have been found to be instrumental in the formation of neurons1,2. However, the instructive role of microRNAs (miRNAs) in neurogenesis remains unexplored. We recently discovered that miR-9* and miR-124 instruct compositional changes of SWI/SNF-like BAF chromatin-remodelling complexes, a process important for neuronal differentiation and function3,4,5,6. Nearing mitotic exit of neural progenitors, miR-9* and miR-124 repress the BAF53a subunit of the neural-progenitor (np)BAF chromatin-remodelling complex. After mitotic exit, BAF53a is replaced by BAF53b, and BAF45a by BAF45b and BAF45c, which are then incorporated into neuron-specific (n)BAF complexes essential for post-mitotic functions4. Because miR-9/9* and miR-124 also control multiple genes regulating neuronal differentiation and function5,7,8,9,10,11,12,13, we proposed that these miRNAs might contribute to neuronal fates. Here we show that expression of miR-9/9* and miR-124 (miR-9/9*-124) in human fibroblasts induces their conversion into neurons, a process facilitated by NEUROD2. Further addition of neurogenic transcription factors ASCL1 and MYT1L enhances the rate of conversion and the maturation of the converted neurons, whereas expression of these transcription factors alone without miR-9/9*-124 was ineffective. These studies indicate that the genetic circuitry involving miR-9/9*-124 can have an instructive role in neural fate determination.

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Figure 1: miRNA-induced transformation of human fibroblasts.
Figure 2: Additional neural factors enhance the conversion to neurons.
Figure 3: Characterization of induced neurons and nBAF subunit expression.
Figure 4: Conversion of adult fibroblasts by miR-9/9*-124-DAM.

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Acknowledgements

We thank I. Graef and A. Cho for helpful suggestions and reagents, A. Kuo and W. Ho for technical help, and X. Bao and P. Khavari for their generous gift of reagents. A.S.Y. is a fellow of the Helen Hay Whitney Foundation. A.X.S. is funded by the Agency of Science, Technology and Research of Singapore (A*STAR). L.L. is supported by the Stanford Medical Scientist Training Program, National Institutes of Mental Health (NIMH) F30MH093125, and the Frances B. Nelson predoctoral fellowship. A.S. is supported by the CIRM post-doctoral fellowship. T.P. is supported by a Swiss National Science Foundation SNSF fellowship for advanced researchers (PA00P3_134196). R.E.D. is supported by the NIH Director’s Award, and awards from the Simon’s Foundation and the CIRM. R.E.D. is also grateful for funding from B. and F. Horowitz, M. McCafferey, B. and J. Packard, P. Kwan and K. Wang. R.W.T. is supported by grants from the Simons, Mathers and Burnett Family Foundations. This work was supported by grants from the Howard Hughes Medical Institute (G.R.C.) and the NIH (HD55391, AI060037 and NS046789 to G.R.C., and NS24067, GM58234 and MH064070 to R.W.T.).

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A.S.Y., A.X.S., and G.R.C. generated the hypotheses and designed experiments. A.S.Y. and A.X.S. performed experiments, generated data in all figures and Supplementary Data. A.S. and L.L. designed and performed experiments for Figs 1, 2 and 4 and Supplementary Data. T.P. designed and performed experiments in Fig. 3a. Y.L. generated data presented in Fig. 1. C.L.-M. performed experiments for Supplementary Data. A.S.Y., A.X.S., L.L., A.S., Y.L., T.P., R.W.T., R.E.D. and G.R.C. wrote the manuscript.

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Correspondence to Andrew S. Yoo or Gerald R. Crabtree.

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The authors declare no competing financial interests.

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Yoo, A., Sun, A., Li, L. et al. MicroRNA-mediated conversion of human fibroblasts to neurons. Nature 476, 228–231 (2011). https://doi.org/10.1038/nature10323

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