Faculty Information |
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Article types | Original article |
Language | English |
Refereed paper | Refereed |
Title | Staurosporine allows dystrophin expression by skipping of nonsense-encoding exon. |
Journal | Formal name:Brain&development Abbreviation:Brain Dev ISSN code:1872-7131(Electronic)0387-7604(Linking) |
Domestic / Foregin | Foregin |
Volume, Number, Page | 38(8),pp.738-45 |
Papers・Author | Nishida Atsushi, Oda Ayaka, Takeuchi Atsuko, Lee Tomoko, Awano Hiroyuki, Hashimoto Naohiro, Takeshima Yasuhiro, Matsuo Masafumi |
Publication date | 2016/09 |
Papers・Description | BACKGROUND:Antisense oligonucleotides that induce exon skipping have been nominated as the most plausible treatment method for dystrophin expression in dystrophin-deficient Duchenne muscular dystrophy. Considering this therapeutic efficiency, small chemical compounds that can enable exon skipping have beenhighly awaited. In our previous report, a small chemical kinase inhibitor, TG003, was shown to enhance dystrophin expression by enhancing exon skipping.PURPOSE:Staurosporine (STS), a small chemical broad kinase inhibitor, was examined for enhanced skipping of a nonsense-encoding dystrophin exon.METHODS:STS was added to culture medium of HeLa cells transfected with minigenes expressing wild-type or mutated exon 31 with c.4303G>T (p.Glu1435X), and the resulting mRNAs were analyzed by RT-PCR amplification. Dystrophin mRNA and protein were analyzed in muscle cells treated with STS by RT-PCR and western blotting, respectively.RESULTS:STS did not alter splicing of the wild-type minigene. In the mutated minigene, STS increased the exon 31-skipped product. A combination of STS and TG003 did not significantly increase the exon 31-skipped product. STS enhanced skipping of exon 4 of the CDC-like kinase 1 gene, whereas TG003 suppressed it. Two STS analogs with selective kinase inhibitory activity did not enhance the mutated exon 31 skipping. When immortalized muscle cells with c.4303G>T in the dystrophin gene were treated with STS, skipping of the mutated exon 31 and dystrophin expression was enhanced.CONCLUSIONS:STS, a broad kinase inhibitor, was shown to enhance skipping of the mutated exon 31 and dystrophin expression, but selective kinase inhibitors did not. |
DOI | 10.1016/j.braindev.2016.03.003 |
PMID | 27021413 |