Protocol for using MYOD1-transduced human urine-derived cells as a predictive platform for exon skipping therapy in Duchenne muscular dystrophy

Katsuhiko Kunitake; Takami Ishizuka; Eri Takeshita; Hirofumi Komaki; Yoshitsugu Aoki

doi:10.1016/j.xpro.2025.103856

Protocol for using MYOD1-transduced human urine-derived cells as a predictive platform for exon skipping therapy in Duchenne muscular dystrophy

Katsuhiko Kunitake
, Takami Ishizuka
, Eri Takeshita
, Hirofumi Komaki
, Yoshitsugu Aoki

研究成果: ジャーナルへの寄稿 › 学術論文 › 査読

抄録

Antisense oligonucleotide (ASO)-based exon skipping is a splice-modulating therapy effective for Duchenne muscular dystrophy (DMD) caused by dystrophin deficiency. Here, we present a protocol for evaluating exon skipping efficacy in MYOD1-transduced human urine-derived cells (MYOD1-UDCs) from patients. We describe steps for isolating UDCs, selecting CD90-positive cells, inducing myogenic differentiation, and assessing the restoration of DMD mRNA and proteins after exon skipping. This platform enhances the predictability of ASO screening, promoting early-stage drug discovery and translational research in DMD. For complete details on the use and execution of this protocol, please refer to Komaki et al.¹

本文言語	英語
論文番号	103856
ジャーナル	STAR Protocols
巻	6
号	2
DOI	https://doi.org/10.1016/j.xpro.2025.103856
出版ステータス	出版済み - 20-06-2025
外部発表	はい

All Science Journal Classification (ASJC) codes

神経科学一般
生化学、遺伝学、分子生物学一般
免疫学および微生物学一般

文献へのアクセス

10.1016/j.xpro.2025.103856

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引用スタイル

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abstract = "Antisense oligonucleotide (ASO)-based exon skipping is a splice-modulating therapy effective for Duchenne muscular dystrophy (DMD) caused by dystrophin deficiency. Here, we present a protocol for evaluating exon skipping efficacy in MYOD1-transduced human urine-derived cells (MYOD1-UDCs) from patients. We describe steps for isolating UDCs, selecting CD90-positive cells, inducing myogenic differentiation, and assessing the restoration of DMD mRNA and proteins after exon skipping. This platform enhances the predictability of ASO screening, promoting early-stage drug discovery and translational research in DMD. For complete details on the use and execution of this protocol, please refer to Komaki et al.1",

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AU - Komaki, Hirofumi

AU - Aoki, Yoshitsugu

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