Abstract
Spinal and bulbar muscular atrophy (SBMA) is an inherited neuromuscular disease caused by expansion of a polyglutamine (polyQ) tract in the androgen receptor (AR). SBMA is triggered by the interaction between polyQ-AR and its natural ligands, testosterone and dihydrotestosterone (DHT). SBMA is characterized by the loss of lower motor neurons and skeletal muscle fasciculations, weakness, and atrophy. To test the hypothesis that the interaction between polyQ-AR and androgens exerts cell-autonomous toxicity in skeletal muscle, we characterized the process of myogenesis and polyQ-AR expression in DHT-treated satellite cells obtained from SBMA patients and age-matched healthy control subjects. Treatment with androgens increased the size and number of myonuclei in myotubes from control subjects, but not from SBMA patients. Myotubes from SBMA patients had a reduced number of nuclei, suggesting impaired myotube fusion and altered contractile structures. The lack of anabolic effects of androgens on myotubes from SBMA patients was not due to defects in myoblast proliferation, differentiation or apoptosis. DHT treatment of myotubes from SBMA patients increased nuclear accumulation of polyQ-AR and decreased the expression of interleukin-4 (IL-4) when compared to myotubes from control subjects. Following DHT treatment, exposure of myotubes from SBMA patients with IL-4 treatment rescued myonuclear number and size to control levels. This supports the hypothesis that androgens alter the fusion process in SBMA myogenesis. In conclusion, these results provide evidence of an androgen-dependent impairment of myogenesis in SBMA that could contribute to disease pathogenesis.
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Acknowledgments
We thank Prof. Egle Perissinotto at University of Padova for statistical analysis, Dr. Emiliano Pena for the collaboration in tissue culture. We are grateful to Prof. Stefano Schiaffino and Dr. Stefano Ciciliot at Venetian Institute of Molecular Medicine for the comments and assistance in discussion, to Dr. Flaviano Favaro and Dr. Diego Faggian at Azienda Ospedale Padova for the kind and efficient collaboration for IL-4 detection. Work supported by Association Française contre les Myopathies (14073 and 14927 to GS, 14631 to LV), Telethon-Italy (GGP10145 to LV; GGP10037 to MP), Progetto d’Ateneo-Università di Padova (to GS). AM was supported by University of Padova, Italy.
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MP received support from Siena Biotech (Italy).
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401_2013_1122_MOESM1_ESM.tif
Supplementary Figure 1. Phosphorylated Akt/Akt ratio in DHT-treated myoblasts and T10 myotubes. (a) Representative WB analysis for phosphorylated Akt (pAkt) and Akt in total cell lysates from DHT-treated myoblasts and T10 myotubes. (b) Ratio of pAkt/total Akt in SBMA myoblasts and T10 myotubes is similar to control. The diagram represents the values expressed as mean ± SD of three independent experiments. The number of single lines studied is given in brackets (TIFF 1180 kb)
401_2013_1122_MOESM2_ESM.tif
Supplementary Figure 2. Apoptotic features in DHT-treated T10 myotubes. (a) Representative images of TUNEL assay. Scale bar, 15 μm. (b) Similar number of TUNEL positive nuclei per myotubes in control and SBMA myotubes; values were obtained in at least 50 myotubes/cell line. The number of single lines studied is given in brackets (TIFF 1724 kb)
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Malena, A., Pennuto, M., Tezze, C. et al. Androgen-dependent impairment of myogenesis in spinal and bulbar muscular atrophy. Acta Neuropathol 126, 109–121 (2013). https://doi.org/10.1007/s00401-013-1122-9
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DOI: https://doi.org/10.1007/s00401-013-1122-9