Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells.
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 (Ca(2+)) influx only in DMD myocytes. Restoration of dystrophin by the exon-skipping technique suppressed this Ca(2+) 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.
Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Emi
E
Shoji
Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Hidetoshi
H
Sakurai
Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Tokiko
T
Nishino
Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Tatsutoshi
T
Nakahata
Department of Paediatrics, Kyoto University Graduate School of Medicine, Kyoto, Japan, 54 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Toshio
T
Heike
Department of Paediatrics, Kyoto University Graduate School of Medicine, Kyoto, Japan, 54 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Tomonari
T
Awaya
Department of Health Promotion Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University, 1-1, Minamiosawa, Hachioji City, Tokyo 192-0397, Japan.
Nobuharu
N
Fujii
Department of Health Promotion Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University, 1-1, Minamiosawa, Hachioji City, Tokyo 192-0397, Japan.
Yasuko
Y
Manabe
The Graduate School of Rehabilitation, Kobe Gakuin University, 518 Arise, Ikawadani-cho, Nishi-ku, Kobe 651-2180, Japan.
Masafumi
M
Matsuo
Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Atsuko
A
Sehara-Fujisawa
D000328:Adult; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D016189:Dystrophin / Q000378:metabolism; D004558:Electric Stimulation; D005091:Exons / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000473:pathology*; D007223:Infant; D008297:Male; D008954:Models, Biological; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000378:metabolism; D020388:Muscular Dystrophy, Duchenne / Q000209:etiology* / Q000473:pathology*; D017570:MyoD Protein / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection
D016189:Dystrophin; D017570:MyoD Protein; D016376:Oligonucleotides, Antisense; D013752:Tetracycline; D002118:Calcium