MicroRNA-mediated conversion of human fibroblasts to neurons (original) (raw)

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

Subjects

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.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

References

  1. Hansen, D. V., Lui, J. H., Parker, P. R. & Kriegstein, A. R. Neurogenic radial glia in the outer subventricular zone of human neocortex. Nature 464, 554–561 (2010)
    Article ADS CAS Google Scholar
  2. Hansen, D. V., Rubenstein, J. L. & Kriegstein, A. R. Deriving excitatory neurons of the neocortex from pluripotent stem cells. Neuron 70, 645–660 (2011)
    Article CAS Google Scholar
  3. Lessard, J. et al. An essential switch in subunit composition of a chromatin remodeling complex during neural development. Neuron 55, 201–215 (2007)
    Article CAS Google Scholar
  4. Wu, J. et al. Regulation of dendritic development by neuron-specific chromatin remodeling complexes. Neuron 56, 94–108 (2007)
    Article CAS Google Scholar
  5. Yoo, A. S., Staahl, B. T., Chen, L. & Crabtree, G. R. MicroRNA-mediated switching of chromatin-remodelling complexes in neural development. Nature 460, 642–646 (2009)
    Article ADS CAS Google Scholar
  6. Wu, J. I., Lessard, J. & Crabtree, G. R. Understanding the words of chromatin regulation. Cell 136, 200–206 (2009)
    Article CAS Google Scholar
  7. Makeyev, E. V., Zhang, J., Carrasco, M. A. & Maniatis, T. The MicroRNA miR-124 promotes neuronal differentiation by triggering brain-specific alternative pre-mRNA splicing. Mol. Cell 27, 435–448 (2007)
    Article CAS Google Scholar
  8. Packer, A. N., Xing, Y., Harper, S. Q., Jones, L. & Davidson, B. L. The bifunctional microRNA miR-9/miR-9* regulates REST and CoREST and is downregulated in Huntington’s disease. J. Neurosci. 28, 14341–14346 (2008)
    Article CAS Google Scholar
  9. Visvanathan, J., Lee, S., Lee, B., Lee, J. W. & Lee, S. K. The microRNA miR-124 antagonizes the anti-neural REST/SCP1 pathway during embryonic CNS development. Genes Dev. 21, 744–749 (2007)
    Article CAS Google Scholar
  10. Cheng, L. C., Pastrana, E., Tavazoie, M. & Doetsch, F. miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche. Nature Neurosci. 12, 399–408 (2009)
    Article CAS Google Scholar
  11. Krichevsky, A. M., Sonntag, K. C., Isacson, O. & Kosik, K. S. Specific microRNAs modulate embryonic stem cell-derived neurogenesis. Stem Cells 24, 857–864 (2006)
    Article CAS Google Scholar
  12. Maiorano, N. A. & Mallamaci, A. Promotion of embryonic cortico-cerebral neuronogenesis by miR-124. Neural Develop. 4, 40 (2009)
    Article Google Scholar
  13. Tang, X. et al. A simple array platform for microRNA analysis and its application in mouse tissues. RNA 13, 1803–1822 (2007)
    Article CAS Google Scholar
  14. Lin, C. H. et al. The dosage of the neuroD2 transcription factor regulates amygdala development and emotional learning. Proc. Natl Acad. Sci. USA 102, 14877–14882 (2005)
    Article ADS CAS Google Scholar
  15. McCormick, M. B. et al. neuroD2 and neuroD3: distinct expression patterns and transcriptional activation potentials within the neuroD gene family. Mol. Cell. Biol. 16, 5792–5800 (1996)
    Article CAS Google Scholar
  16. Olson, J. M. et al. NeuroD2 is necessary for development and survival of central nervous system neurons. Dev. Biol. 234, 174–187 (2001)
    Article CAS Google Scholar
  17. Ince-Dunn, G. et al. Regulation of thalamocortical patterning and synaptic maturation by NeuroD2. Neuron 49, 683–695 (2006)
    Article CAS Google Scholar
  18. Ryan, T. A. et al. The kinetics of synaptic vesicle recycling measured at single presynaptic boutons. Neuron 11, 713–724 (1993)
    Article CAS Google Scholar
  19. Vierbuchen, T. et al. Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463, 1035–1041 (2010)
    Article ADS CAS Google Scholar
  20. Parrish, J. Z., Kim, M. D., Jan, L. Y. & Jan, Y. N. Genome-wide analyses identify transcription factors required for proper morphogenesis of Drosophila sensory neuron dendrites. Genes Dev. 20, 820–835 (2006)
    Article CAS Google Scholar
  21. Ho, L. et al. An embryonic stem cell chromatin remodeling complex, esBAF, is an essential component of the core pluripotency transcriptional network. Proc. Natl Acad. Sci. USA 106, 5187–5191 (2009)
    Article ADS CAS Google Scholar
  22. Ho, L., Miller, E. L., Ronan, J. L., Ho, W. Q., Jothi, R. & Crabtree, G. R. esBAF facilitates pluripotency by conditioning the genome for LIF/STAT3 signaling and by regulating polycomb function. Nature Cell Biol. (in the press)
  23. Coolen, M. & Bally-Cuif, L. MicroRNAs in brain development and physiology. Curr. Opin. Neurobiol. 19, 461–470 (2009)
    Article CAS Google Scholar
  24. Wu, J. & Xie, X. Comparative sequence analysis reveals an intricate network among REST, CREB and miRNA in mediating neuronal gene expression. Genome Biol. 7, R85 (2006)
    Article Google Scholar
  25. Laneve, P. et al. A minicircuitry involving REST and CREB controls miR-9-2 expression during human neuronal differentiation. Nucleic Acids Res. 38, 6895–6905 (2010)
    Article CAS Google Scholar
  26. Andres, M. E. et al. CoREST: a functional corepressor required for regulation of neural-specific gene expression. Proc. Natl Acad. Sci. USA 96, 9873–9878 (1999)
    Article ADS CAS Google Scholar
  27. Barry, P. H. JPCalc, a software package for calculating liquid junction potential corrections in patch-clamp, intracellular, epithelial and bilayer measurements and for correcting junction potential measurements. J. Neurosci. Methods 51, 107–116 (1994)
    Article ADS CAS Google Scholar

Download references

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
    You can also search for this author inPubMed Google Scholar
  2. Alfred X. Sun
    You can also search for this author inPubMed Google Scholar
  3. Li Li
    You can also search for this author inPubMed Google Scholar
  4. Aleksandr Shcheglovitov
    You can also search for this author inPubMed Google Scholar
  5. Thomas Portmann
    You can also search for this author inPubMed Google Scholar
  6. Yulong Li
    You can also search for this author inPubMed Google Scholar
  7. Chris Lee-Messer
    You can also search for this author inPubMed Google Scholar
  8. Ricardo E. Dolmetsch
    You can also search for this author inPubMed Google Scholar
  9. Richard W. Tsien
    You can also search for this author inPubMed Google Scholar
  10. Gerald R. Crabtree
    You can also search for this author inPubMed Google Scholar

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 toAndrew S. Yoo or Gerald R. Crabtree.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

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

Download citation

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.

Associated content