Dystrophin expression in muscle stem cells regulates their polarity and asymmetric division

Abstract

Dystrophin is expressed in differentiated myofibers, in which it is required for sarcolemmal integrity, and loss-of-function mutations in the gene that encodes it result in Duchenne muscular dystrophy (DMD), a disease characterized by progressive and severe skeletal muscle degeneration. Here we found that dystrophin is also highly expressed in activated muscle stem cells (also known as satellite cells), in which it associates with the serine-threonine kinase Mark2 (also known as Par1b), an important regulator of cell polarity. In the absence of dystrophin, expression of Mark2 protein is downregulated, resulting in the inability to localize the cell polarity regulator Pard3 to the opposite side of the cell. Consequently, the number of asymmetric divisions is strikingly reduced in dystrophin-deficient satellite cells, which also display a loss of polarity, abnormal division patterns (including centrosome amplification), impaired mitotic spindle orientation and prolonged cell divisions. Altogether, these intrinsic defects strongly reduce the generation of myogenic progenitors that are needed for proper muscle regeneration. Therefore, we conclude that dystrophin has an essential role in the regulation of satellite cell polarity and asymmetric division. Our findings indicate that muscle wasting in DMD not only is caused by myofiber fragility, but also is exacerbated by impaired regeneration owing to intrinsic satellite cell dysfunction.

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Figure 1: Dystrophin expression in satellite cells.
Figure 2: Impaired satellite stem cell asymmetric divisions and reduced generation of myogenic progenitors in the absence of dystrophin.
Figure 3: Dystrophin regulates PAR polarity protein localization.
Figure 4: PAR polarity proteins are required for muscle stem cell asymmetric divisions.
Figure 5: Dystrophin-deficient satellite cells display impaired mitotic spindle orientation and loss of apicobasal division.
Figure 6: Dystrophin-deficient satellite cells have reduced ability to generate myogenic progenitors in regenerating muscle.

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Acknowledgements

We thank J. Dilworth and L. Megeney for careful reading of the manuscript. We also thank J. Ritchie for animal husbandry, and J. Fernandes and P. Oleynik of the flow cytometry facility of the StemCore laboratories for technical assistance. N.A.D. is supported by a Postdoctoral Fellowship from the Canadian Institutes of Health Research (CIHR); Y.X.W. is supported by fellowships from the Queen Elizabeth II Graduate Scholarships in Science and Technology and the CIHR; J.v.M. was supported by a grant from the Deutsche Forschungsgemeinschaft; C.F.B. was supported by a grant from the Swiss National Science Foundation; C.E.B. is supported by a Postdoctoral Fellowship from the Ontario Institute for Regenerative Medicine; and M.A.R. holds the Canada Research Chair in Molecular Genetics. These studies were carried out with support from grants to M.A.R. from the US National Institutes for Health (grant no. RO1AR044031), the CIHR (grant no. MOP-12080 and MOP-81288), the E-Rare-2 program from the CIHR and Muscular Dystrophy Canada (grant no. ERA-132935), the Muscular Dystrophy Association, the Stem Cell Network and the Ministry of Research and Innovation (MRI), Government of Ontario (grant no. ORF-RE05-084).

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N.A.D. and Y.X.W. designed and carried out experiments, analyzed results and wrote the manuscript. J.v.M. designed and conducted experiments and analyzed results. A.P., C.F.B. and C.E.B. conducted experiments. M.A.R. designed experiments, analyzed results, wrote the manuscript and provided financial support.

Correspondence to Michael A Rudnicki.

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Dumont, N., Wang, Y., von Maltzahn, J. et al. Dystrophin expression in muscle stem cells regulates their polarity and asymmetric division. Nat Med 21, 1455–1463 (2015). https://doi.org/10.1038/nm.3990

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