Abstract
Duchenne muscular dystrophy (DMD) is a progressive and fatal muscle degenerating disease caused by a dystrophin deficiency. Effective suppression of the primary pathology observed in DMD is critical for treatment. Patient-derived human induced pluripotent stem cells (hiPSCs) are a promising tool for drug discovery. Here, we report an in vitro evaluation system for a DMD therapy using hiPSCs that recapitulate the primary pathology and can be used for DMD drug screening. Skeletal myotubes generated from hiPSCs are intact, which allows them to be used to model the initial pathology of DMD in vitro. Induced control and DMD myotubes were morphologically and physiologically comparable. However, electric stimulation of these myotubes for in vitro contraction caused pronounced calcium ion (Ca2+) influx only in DMD myocytes. Restoration of dystrophin by the exon-skipping technique suppressed this Ca2+ overflow and reduced the secretion of creatine kinase (CK) in DMD myotubes. These results suggest that the early pathogenesis of DMD can be effectively modelled in skeletal myotubes induced from patient-derived iPSCs, thereby enabling the development and evaluation of novel drugs.
Highlights
Duchenne muscular dystrophy (DMD) is a progressive and fatal muscle degenerating disease caused by a dystrophin deficiency
Skin fibroblasts were biopsied from 2 different patients who were diagnosed with DMD and had deletion of exon 44 (Δ 44) and exon 46–47 (Δ 46–47) in DMD, respectively, and from the 36-year-old biological father of the Δ 44-patient
We have recapitulated the early pathogenesis of DMD by creating an in vitro DMD model with myotubes generated from patient-derived iPS cells
Summary
Duchenne muscular dystrophy (DMD) is a progressive and fatal muscle degenerating disease caused by a dystrophin deficiency. Relative mRNA expression at the end of the differentiation process at day 9 indicated that the differentiated skeletal muscle cells from DMD-iPStet-MyoD and Control-Myocytes were comparable (Fig. 2b). Immunocytochemistry demonstrated that both Control- and DMD-Myocytes differentiated to form myotubes that express myosin heavy chain (MHC), CKM, and skeletal muscle actin (SMA) at day 9 of induction (Fig. 2c).
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