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Transcription factor–mediated reprogramming of fibroblasts to expandable, myelinogenic oligodendrocyte progenitor cells

Nature Biotechnology volume 31, pages 426–433 (2013)Cite this article

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Abstract

Cell-based therapies for myelin disorders, such as multiple sclerosis and leukodystrophies, require technologies to generate functional oligodendrocyte progenitor cells. Here we describe direct conversion of mouse embryonic and lung fibroblasts to induced oligodendrocyte progenitor cells (iOPCs) using sets of either eight or three defined transcription factors. iOPCs exhibit a bipolar morphology and global gene expression profile consistent with bona fide OPCs. They can be expanded in vitro for at least five passages while retaining the ability to differentiate into multiprocessed oligodendrocytes. When transplanted to hypomyelinated mice, iOPCs are capable of ensheathing host axons and generating compact myelin. Lineage conversion of somatic cells to expandable iOPCs provides a strategy to study the molecular control of oligodendrocyte lineage identity and may facilitate neurological disease modeling and autologous remyelinating therapies.

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Figure 1: Eight TFs reprogram MEFs to induced OPCs.
Figure 2: Eight TF-induced MEFs exhibit properties of bona fide OPCs.
Figure 3: Eight TF-induced MEFs function to generate compact myelin.
Figure 4: A2B5 immunosorting allows for the prospective enrichment of iOPCs.
Figure 5: Sox10, Olig2 and Nkx6.2 are sufficient to reprogram fibroblasts to iOPCs.

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Gene Expression Omnibus

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Gene Expression Omnibus

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Acknowledgements

This research was supported by US National Institutes of Health (NIH) grants MH087877 (P.J.T. and R.H.M.), NS30800 (R.H.M.) and EY019880 (T.M.); NIH predoctoral training grants F31NS083354 (A.M.L.) and T32GM008056 (R.T.K.); Case Western Reserve University School of Medicine (P.J.T.); the New York Stem Cell Foundation (P.J.T.); the Mt. Sinai Health Care Foundation (P.J.T.); and the Cytometry & Imaging Microscopy, and the Gene Expression and Genotyping core facilities of the Case Comprehensive Cancer Center (P30CA043703). P.J.T. is a New York Stem Cell Foundation–Robertson Investigator. We are grateful to L. Cooperman, J. Wanta, M. Hitomi, J. Krasno, M. Karl, S. Fyffe-Maricich, S. Wu, M. Pendergast, O. Corradin, T. LaFramboise and P. Scacheri for technical assistance, I. Tsung for artwork, as well as N. Avishai and G. Kidd for 3D scanning electron microscopy imaging and data processing.

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Author notes

  1. Fadi J Najm and Angela M Lager: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics and Genome Sciences, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    Fadi J Najm, Angela M Lager, Daniel C Factor, Robert T Karl & Paul J Tesar

  2. Department of Neurosciences, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    Anita Zaremba, Krysta Wyatt, Andrew V Caprariello, Robert H Miller & Paul J Tesar

  3. Department of Ophthalmology & Visual Sciences, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    Tadao Maeda

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Contributions

F.J.N., A.M.L. and P.J.T. designed the reprogramming strategy and generated all iOPCs; F.J.N., A.M.L., R.T.K. and P.J.T. performed in vitro differentiation experiments; A.V.C., A.Z., F.J.N., A.M.L. and T.M. performed in vivo myelination experiments; A.Z., F.J.N., A.M.L., R.H.M. and P.J.T. performed slice culture myelination experiments; K.W., A.Z. and R.H.M. produced electron microscope images; F.J.N., A.M.L., D.C.F. and P.J.T. generated and analyzed gene expression data; F.J.N., A.M.L., R.H.M. and P.J.T. analyzed all of the data and wrote the paper. All authors edited and approved the final manuscript.

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Correspondence to Paul J Tesar.

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Competing interests

P.J.T., R.H.M. and F.J.N. have a pending patent application for this technology.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 (PDF 7874 kb)

Supplementary Table 1

Transcription factors enriched in CNS lineages (used to generate Figure 1b) (XLS 21 kb)

Supplementary Table 2

GREAT analysis for Najm, Lager, et al. (2013) (XLS 186 kb)

Supplementary Movie 1

3-dimensional reconstruction of serial block-face scanning electron micrographs showing iOPC myelin in the dorsal column of the hypomyelinated shiverer spinal cord (MOV 30996 kb)

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Najm, F., Lager, A., Zaremba, A. et al. Transcription factor–mediated reprogramming of fibroblasts to expandable, myelinogenic oligodendrocyte progenitor cells. Nat Biotechnol 31, 426–433 (2013). https://doi.org/10.1038/nbt.2561

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