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MicroRNA-mediated conversion of human fibroblasts to neurons - Nature

  • ️Crabtree, Gerald R.
  • ️Wed Jul 13 2011
  • Letter
  • Published: 13 July 2011

Nature volume 476, pages 228–231 (2011)Cite this article

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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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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.).

Author information

Author notes

  1. Andrew S. Yoo

    Present address: Present address: Department of Developmental Biology, Washington University in St Louis, St Louis, Missouri 63110, USA.,

  2. Andrew S. Yoo, Alfred X. Sun, Li Li and Aleksandr Shcheglovitov: These authors contributed equally to this work.

Authors and Affiliations

  1. Howard Hughes Medical Institute and the Departments of Developmental Biology and of Pathology, Stanford University, Stanford, 94305, California, USA

    Andrew S. Yoo & Gerald R. Crabtree

  2. Program in Cancer Biology, Stanford University, Stanford, 94305, California, USA

    Alfred X. Sun

  3. Department of Molecular and Cellular Physiology, Stanford University, Stanford, 94305, California, USA

    Li Li, Yulong Li & Richard W. Tsien

  4. Medical Scientist Training Program, Stanford University, Stanford, 94305, California, USA

    Li Li

  5. Neuroscience Program, Stanford University, Stanford, 94305, California, USA

    Li Li

  6. Department of Neurobiology, Stanford University, Stanford, 94305, California, USA

    Aleksandr Shcheglovitov, Thomas Portmann & Ricardo E. Dolmetsch

  7. Department of Neurology, Stanford University, Stanford, 94305, California, USA

    Chris Lee-Messer

Authors

  1. Andrew S. Yoo

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  2. Alfred X. Sun

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  3. Li Li

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  4. Aleksandr Shcheglovitov

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  5. Thomas Portmann

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  6. Yulong Li

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  7. Chris Lee-Messer

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  8. Ricardo E. Dolmetsch

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  9. Richard W. Tsien

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  10. Gerald R. Crabtree

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Contributions

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.

Corresponding authors

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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  • Received: 21 July 2010

  • Accepted: 27 June 2011

  • Published: 13 July 2011

  • Issue Date: 11 August 2011

  • DOI: https://doi.org/10.1038/nature10323

Editorial Summary

Neurons from fibroblasts

Three papers in this issue demonstrate the production of functional induced neuronal (iN) cells from human fibroblasts, a procedure that holds great promise for regenerative medicine. Pang et al. show that a combination of the three transcription factors Ascl1 (also known as Mash1), Brn2 (or Pou3f2) and Myt1l greatly enhances the neuronal differentiation of human embryonic stem cells. When combined with the basic helix–loop–helix transcription factor NeuroD1, these factors can also convert fetal and postnatal human fibroblasts into iN cells. Caiazzo et al. use a cocktail of three transcription factors to convert prenatal and adult mouse and human fibroblasts into functional dopaminergic neurons. The three are Mash1, Nurr1 (or Nr4a2) and Lmx1a. Conversion is direct with no reversion to a progenitor cell stage, and it occurs in cells from Parkinson's disease patients as well as from healthy donors. Yoo et al. use an alternative approach. They show that microRNAs can have an instructive role in neural fate determination. Expression of miR-9/9* and miR-124 in human fibroblasts induces their conversion into functional neurons, and the process is facilitated by the addition of some neurogenic transcription factors.

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