Abstract
Myotonic dystrophy type 1 (DM1) is caused by CTG nucleotide repeat expansions in the 3′-untranslated region (3′-UTR) of the dystrophia myotonica protein kinase (DMPK) gene. The expanded CTG repeats encode toxic CUG RNAs that cause disease, largely through RNA gain-of-function. DM1 is a fatal disease characterized by progressive muscle wasting, which has no cure. Regenerative medicine has emerged as a promising therapeutic modality for DM1, especially with the advancement of induced pluripotent stem (iPS) cell technology and therapeutic genome editing. However, there is an unmet need to identify in vitro outcome measures to demonstrate the therapeutic effects prior to in vivo clinical trials. In this study, we examined the muscle regeneration (myotube formation) in normal and DM1 myoblasts in vitro to establish outcome measures for therapeutic monitoring. We found normal proliferation of DM1 myoblasts, but abnormal nuclear aggregation during the early stage myotube formation, as well as myotube degeneration during the late stage of myotube formation. We concluded that early abnormal nuclear aggregation and late myotube degeneration offer easy and sensitive outcome measures to monitor therapeutic effects in vitro.
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Acknowledgements
Research reported in this publication was supported by the NIH/NIAMS grant K08 AR064836 to G.X. mentored by Maury S. Swanson, Tetsuo Ashizawa, Laura Ranum, Naohiro Terada, and SH Subramony at the University of Florida and Fernando Valenzuela in University of New Mexico and by startup funds from the Office of Research at the Health Science Center of University of New Mexico. We would like to thank Alexis K Hall for proofreading, as well as submitting suggestions on the manuscript.
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Supplemental Fig. 1
Homogeneity of myoblasts in normal and DM1 myoblasts passage 7 (lower power images). (PNG 1334 kb)
Supplemental Fig. 2
Nuclear aggregation in DM1 myotubes. On day 3, the nuclear aggregation was easily spotted in DM1 myotubes (B) while such phenomena were barely seen in normal myotubes (phase-contrast images). (PNG 201 kb)
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Wang, Y., Hao, L., Li, H. et al. Abnormal nuclear aggregation and myotube degeneration in myotonic dystrophy type 1. Neurol Sci 40, 1255–1265 (2019). https://doi.org/10.1007/s10072-019-03783-w
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DOI: https://doi.org/10.1007/s10072-019-03783-w