Abstract

Oligodendrocytes wrap nerve fibres in the central nervous system with layers of specialized cell membrane to form myelin sheaths1. Myelin is destroyed by the immune system in multiple sclerosis, but myelin is thought to regenerate and neurological function can be recovered. In animal models of demyelinating disease, myelin is regenerated by newly generated oligodendrocytes, and remaining mature oligodendrocytes do not seem to contribute to this process2,3,4. Given the major differences in the dynamics of oligodendrocyte generation and adaptive myelination between rodents and humans5,6,7,8,9, it is not clear how well experimental animal models reflect the situation in multiple sclerosis. Here, by measuring the integration of 14C derived from nuclear testing in genomic DNA10, we assess the dynamics of oligodendrocyte generation in patients with multiple sclerosis. The generation of new oligodendrocytes was increased several-fold in normal-appearing white matter in a subset of individuals with very aggressive multiple sclerosis, but not in most subjects with the disease, demonstrating an inherent potential to substantially increase oligodendrocyte generation that fails in most patients. Oligodendrocytes in shadow plaques—thinly myelinated lesions that are thought to represent remyelinated areas—were old in patients with multiple sclerosis. The absence of new oligodendrocytes in shadow plaques suggests that remyelination of lesions occurs transiently or not at all, or that myelin is regenerated by pre-existing, and not new, oligodendrocytes in multiple sclerosis. We report unexpected oligodendrocyte generation dynamics in multiple sclerosis, and this should guide the use of current, and the development of new, therapies.

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Data availability

Source Data for Figs. 1d, 24 and Extended Data Figs. 46, 7e, f, 8 have been provided. All other data supporting the findings of this study are available from the corresponding author upon reasonable request.

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Change history

  • 05 February 2019

    In this Letter, the vertical error bars were missing from Fig. 3b and 3c. This figure has been corrected online.

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Acknowledgements

We thank S. Giatrellis and M. Toro for flow cytometry and K. Håkansson and P. Senneryd for accelerator mass spectrometry sample preparation. This study was supported by the Swedish Research Council, the Swedish Cancer Foundation, Tobias Stiftelsen, SSF, Knut och Alice Wallenbergs Stiftelse, the ERC and Torsten Söderberg Foundation. Tissue samples and associated clinical and neuropathological data were supplied by the Multiple Sclerosis Society Tissue Bank, funded by the Multiple Sclerosis Society of Great Britain and Northern Ireland, registered charity 207495. We also thank B. D. Trapp and R. Dutta, Lerner Research Institute, Cleveland Clinic, Cleveland, USA and the Netherlands Brain Bank, Netherlands Institute for Neuroscience, Amsterdam, for providing tissue.

Reviewer information

Nature thanks K. Nave and the anonymous reviewer(s) for their contribution to the peer review of this work.

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Affiliations

  1. Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden

    • Maggie S. Y. Yeung
    • , Mehdi Djelloul
    • , Embla Steiner
    •  & Jonas Frisén
  2. Institut Camille Jordan, CNRS UMR 5208, University of Lyon, Villeurbanne, France

    • Samuel Bernard
  3. Department of Physics and Astronomy, Ion Physics, Uppsala University, Uppsala, Sweden

    • Mehran Salehpour
    •  & Göran Possnert
  4. Department of Clinical Neuroscience, Division of Neurology, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden

    • Lou Brundin

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Contributions

M.S.Y.Y. and M.D. performed the experiments. M.S.Y.Y. and E.S. performed statistical analysis. E.S. and S.B. performed mathematical modelling. M.S. and G.P. performed 14C measurements. M.S.Y.Y. and J.F. designed the study and experiments. M.S.Y.Y., J.F. and L.B. designed data analysis. E.S., S.B., M.S.Y.Y. and J.F. designed mathematical analysis. J.F. supervised the study. M.S.Y.Y., E.S. and J.F. prepared figures and wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Jonas Frisén.

Extended data figures and tables

  1. Extended Data Fig. 1 Flow cytometry reanalysis of isolated cell nuclei from patients with multiple sclerosis.

    ac, Reanalysis of mature oligodendrocytes, OPCs and non-oligodendrocyte lineage cells isolated by flow cytometry shows 94.8 ± 2.1%, 82.2 ± 10.2% and 97.6 ± 2.1% purity, respectively (n = 29, 24 and 28 biopsies and patients, for each analysis, 20,000 events were recorded per sorted fraction, technical replicates per fraction (≥3)), and the respective contamination of OPCs (top left quadrant), non-oligodendrocyte linage cells (bottom left quadrant) and mature oligodendrocyte (top right quadrant) in the isolated population. Values inside the quadrants represent the average percentage from all isolated NAWM of patients with multiple sclerosis (mean ± s.d.). The small fraction of contamination of other than the isolated population acquired during flow cytometry isolation was accounted for by correcting the 14C values to reflect the isolated cell population accurately. See also Method and Supplementary Table 1.

  2. Extended Data Fig. 2 Visualization of myelin in shadow plaques.

    a, Overview of a carbon-dated multiple sclerosis tissue block reveals a shadow plaque and NAWM by eriochrome cyanine staining (n = 11). The shadow plaque is outlined by a dashed line. bd, Higher magnification views of areas indicated by boxes in a show reduced myelin (proteolipid protein (PLP)) in all areas compared to NAWM. The images show patient case MS10. See also Supplementary Table 1. Scale bars, 2 mm (a) and 20 μm (bd).

  3. Extended Data Fig. 3 Reduced myelin in carbon-dated multiple sclerosis shadow plaques.

    Reduced myelin staining (PLP) was observed in all shadow plaque areas (border and centre of shadow plaque) compared to NAWM. NAWM and shadow plaque images from each patient were taken from the same sample tissue block (n = 11). See also Supplementary Table 1. Scale bar, 20 μm.

  4. Extended Data Fig. 4 Modelled 14C concentration in DNA of oligodendrocytes generated until disease onset.

    The modelled 14C concentration for each patient born before the nuclear bomb tests with dated mature oligodendrocytes from a shadow plaque, if there would have been normal oligodendrocyte generation dynamics until the time of disease onset and no further generation thereafter. The measured data from each patient is shown as a blue circle, and the modelled data for each patient in purple (n = 10). The red line depicts genomic DNA 14C concentrations in oligodendrocytes from healthy subjects. Source data

  5. Extended Data Fig. 5 Modelling of the highest possible turnover during the disease period that could occur undetected with carbon dating.

    Different annual turnover rates during the disease period were modelled to find the highest possible turnover rate that could occur undetected. It was assumed that there was a healthy turnover before disease onset. If a comparison between the measured 14C and the modelled values is significant, we would be able to detect that turnover rate. The lowest turnover rate with significant difference tested was 0.1% annual turnover (P = 0.041, two-tailed Mann–Whitney U-test, n = 6). At 0.05% annual turnover, there was no longer a significant difference (P = 0.132, two-tailed Mann–Whitney U-test, n = 6). Data are mean (long line), ± s.d. (short lines). Source data

  6. Extended Data Fig. 6 Oligodendrocyte density in shadow plaques.

    Pairwise comparison of the density of SOX10+NOGO-A+ mature oligodendrocytes in histological sections of shadow plaques and adjacent NAWM in the same patients (n = 11 biopsies from 11 patients). Source data

  7. Extended Data Fig. 7 Cell proliferation in multiple sclerosis.

    ad, Proliferating cells (Ki-67+, arrowheads, ac) are sparse and proliferating OPCs are very rare (Ki-67+SOX10+, open arrowhead, d) in histological sections of shadow plaques (ac) and adjacent NAWM (d) from patients with multiple sclerosis (n = 11). The box in a indicates the area shown in higher magnification (b) and the orthogonal view (c). Scale bars, 100 µm (a) and 10 µm (bd). e, f, Quantification of Ki-67+ cells (e) and Ki-67+SOX10+ cells (f) in shadow plaque and adjacent NAWM in patients with multiple sclerosis (n = 11). Source data

  8. Extended Data Fig. 8 Modelled 14C concentration in DNA of oligodendrocytes generated from OPCs in shadow plaques.

    a, Measured and modelled data. The levels of atmospheric 14C before 1955 are indicated by the dotted line. Mean (long line) ± s.d. (short lines). Statistical analyses represent comparison between shadow plaque data (n = 6) and the other groups (model 1–7, see bh) compared to healthy white matter (**P = 0.0047, two-tailed Mann–Whitney U-test, n = 10). bh, Measured and modelled data plotted in relation to time and atmospheric 14C concentration. b, Model 1: the estimated 14C concentration in mature oligodendrocytes in shadow plaques if assumed to have healthy turnover rate until the death of the patients is similar to healthy white matter oligodendrocytes (red line and red circles in a) and is significantly higher than the measured 14C levels in mature oligodendrocytes in shadow plaques (in a, **P = 0.002, two-tailed Mann–Whitney U-test, n = 6 and 6). c, d, Models 2 and 3: to reconstitute the observed mature oligodendrocyte density with new oligodendrocytes, it would require OPCs to divide at least four times. If this occurred over the time course from the onset of the disease (model 2; c) or even from 10 years before the first clinical symptoms (model 3; d) until the death of the patient, the resulted modelled 14C levels were significantly higher than the 14C levels measured from mature oligodendrocytes in shadow plaques (in a, model 2, **P = 0.002, two-tailed Mann–Whitney U-test, n = 6, model 3, **P = 0.002, two-tailed Mann–Whitney U-test, n = 6). e, f, Models 4 and 5: even with more conservative models that assume all oligodendrocyte replacements occur not throughout the disease course, but early by the proliferation of OPCs at disease onset or before, the modelled 14C levels still result in higher levels than the measured shadow plaque oligodendrocytes (blue) in some patients (a). g, h, Models 6 and 7: modelled data as above, but with 50% of oligodendrocytes being newly made from OPCs and 50% being old oligodendrocytes. For some patients, the time of disease onset and 10 years before onset occur before the rise of the 14C atmospheric levels (1955). For these patients, any cell replacement by the proliferation of old OPCs during this period would results in values similar to atmospheric levels. Arrows points to patients with time periods before and disease onsets that overlap with very highly increased atmospheric 14C levels compared to respective levels at time of birth of the patients, and that the modelled values deviate from the measured rules out these scenarios. The red line depicts genomic DNA 14C concentrations in oligodendrocytes from healthy subjects. **P < 0.01. See also Supplementary Tables 1, 11 and 12. Source data

Supplementary information

  1. Supplementary Information

    This file contains Supplementary Tables 1-12 and an analysis of cell dynamics in multiple sclerosis white matter

  2. Reporting Summary

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https://doi.org/10.1038/s41586-018-0842-3

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