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Abstract

Reprogramming somatic cells from one cell fate to another can generate specific neurons suitable for disease modeling. To maximize the utility of patient-derived neurons, they must model not only disease-relevant cell classes, but also the diversity of neuronal subtypes found in vivo and the pathophysiological changes that underlie specific clinical diseases. We identified five transcription factors that reprogram mouse and human fibroblasts into noxious stimulus–detecting (nociceptor) neurons. These recapitulated the expression of quintessential nociceptor-specific functional receptors and channels found in adult mouse nociceptor neurons, as well as native subtype diversity. Moreover, the derived nociceptor neurons exhibited TrpV1 sensitization to the inflammatory mediator prostaglandin E2 and the chemotherapeutic drug oxaliplatin, modeling the inherent mechanisms underlying inflammatory pain hypersensitivity and painful chemotherapy-induced neuropathy. Using fibroblasts from patients with familial dysautonomia (hereditary sensory and autonomic neuropathy type III), we found that the technique was able to reveal previously unknown aspects of human disease phenotypes in vitro.

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Change history

  • 10 December 2014

    In the version of this article initially published online, an affilation was missing for author Julia T. Oliveira: Laboratório de Neurodegeneração e Reparo, Departamento de Patologia, Faculdade de Medicina, Universidade Federal Do Rio De Janeiro, Rio de Janeiro, Brazil. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank M. Costigan for assistance with RT-PCR, A. Yekkirala and J. Sprague for help with calcium imaging, Q. Ma (Dana-Farber Cancer Institute) and E. Turner (Seattle Children's Research Institute) for constructs, J. Gardner and J. McNeish for helpful advice and support, and K. Wainger for assistance with figure preparation. We also thank the Boston Children's Hospital IDDRC Molecular Genetics Core Facility for RNA Bioanalyzer analyses and the Harvard Medical School ICCB Screening Facility for assistance with ImageXpress and MetaXpress analyses. This research was supported by the National Institute of General Medical Sciences (T32 GM07592) and National Institute of Neurological Disorders and Stroke (1K08-NS082364) to B.J.W. Conselho Nacional de Desenvolvimento Científico e Tecnológico (J.T.O.), GlaxoSmithKline Regenerative Medicine DPU (C.J.W.), the National Institute of Neurological Disorders and Stroke (NS038253 to C.J.W.), and the Dr. Miriam and Sheldon G. Adelson Medical Foundation (C.J.W.).

Author information

Author notes

    • Brian J Wainger
    •  & Elizabeth D Buttermore

    These authors contributed equally to this work.

Affiliations

  1. F.M. Kirby Neurobiology Center, Boston Children's Hospital and Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.

    • Brian J Wainger
    • , Elizabeth D Buttermore
    • , Julia T Oliveira
    • , Cassidy Mellin
    • , Seungkyu Lee
    • , Wardiya Afshar Saber
    • , Amy J Wang
    • , Isaac M Chiu
    • , Lee Barrett
    • , Eric A Huebner
    • , Canan Bilgin
    • , Christian Brenneis
    • , Kush Kapur
    •  & Clifford J Woolf
  2. Department of Anesthesia, Critical Care and Pain Medicine and Department of Neurology, Massachusetts General Hospital, Boston, Massachusetts, USA.

    • Brian J Wainger
  3. Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.

    • Brian J Wainger
    • , Elizabeth D Buttermore
    • , Seungkyu Lee
    • , Isaac M Chiu
    • , Eric A Huebner
    •  & Clifford J Woolf
  4. Laboratório de Neurodegeneração e Reparo, Departamento de Patologia, Faculdade de Medicina, Universidade Federal Do Rio De Janeiro, Rio de Janeiro, Brazil.

    • Julia T Oliveira
  5. Harvard Stem Cell Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.

    • Justin K Ichida
    • , Naomi Tsujimoto
    • , Lee L Rubin
    •  & Kevin Eggan
  6. Department of Stem Cell Biology and Regenerative Medicine, Eli and Edythe Broad CIRM Center for Regenerative Medicine and Stem Cell Research, University of Southern California, Los Angeles, California, USA.

    • Justin K Ichida
  7. The Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.

    • Kevin Eggan

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Contributions

B.J.W. conceived, designed and performed the lineage reprogramming experiments and physiological experiments, analyzed data, and wrote the manuscript. E.D.B. designed, performed and analyzed reprogramming, quantitative PCR, single-cell RT-PCR, immunohistochemistry and CGRP ELISA experiments, and wrote the manuscript. J.T.O. performed and optimized the induced nociceptor technique. C.M. performed and analyzed physiological studies and edited the manuscript. S.L. performed CGRP ELISA and single-cell RT-PCR assays. W.A.S. performed reprogramming and immunohistochemistry experiments. A.J.W. performed initial cloning and transduction experiments. J.K.I. provided essential advice for nociceptor reprogramming strategy and edited the manuscript. I.M.C. gave critical advice regarding the genetic reporter, choice of transcription factors, performed cell sorting experiments and edited the manuscript. L.B. advised and performed molecular biology experiments. E.A.H. performed image quantification and analysis. C. Bilgin assisted with reprogramming and immunohistochemistry. N.T. assisted with human motor neuron culture and, together with C. Brenneis performed culture and characterization using initial approaches. K.K. performed statistical modeling of human nociceptor data. L.L.R. advised regarding reprogramming experiments and edited the manuscript. K.E. provided advice and reagents for reprogramming and edited the manuscript. C.J.W. designed experiments, interpreted findings and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Clifford J Woolf.

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https://doi.org/10.1038/nn.3886

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