論文

査読有り 本文へのリンクあり 国際誌
2022年11月

Antisense oligonucleotide induced pseudoexon skipping and restoration of functional protein for Fukuyama muscular dystrophy caused by a deep-intronic variant

Human Molecular Genetics
  • Sarantuya Enkhjargal
  • ,
  • Kana Sugahara
  • ,
  • Behnoush Khaledian
  • ,
  • Miwako Nagasaka
  • ,
  • Hidehito Inagaki
  • ,
  • Hiroki Kurahashi
  • ,
  • Hisatsugu Koshimizu
  • ,
  • Tatsushi Toda
  • ,
  • Mariko Taniguchi-Ikeda

32
8
開始ページ
1301
終了ページ
1312
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1093/hmg/ddac286
出版者・発行元
Oxford University Press

Fukuyama congenital muscular dystrophy (FCMD) is an autosomal recessive disorder caused by fukutin (FKTN) gene mutations. FCMD is the second most common form of childhood muscular dystrophy in Japan, and the most patients possess a homozygous retrotransposal SINE-VNTR-Alu insertion in the 3′-untranslated region of FKTN. A deep-intronic variant (DIV) was previously identified as the second most prevalent loss-of-function mutation in Japanese patients with FCMD. The DIV creates a new splicing donor site in intron 5 that causes aberrant splicing and the formation of a 64-base pair pseudoexon in the mature mRNA, resulting in a truncated nonfunctional protein. Patients with FCMD carrying the DIV present a more severe symptoms, and currently, there is no radical therapy available for this disorder. In the present study, we describe in vitro evaluation of antisense oligonucleotide mediated skipping of pseudoexon inclusion and restoration of functional FKTN protein. A total of 16 19-26-mer antisense oligonucleotide sequences were designed with a 2’-O-methyl backbone and were screened in patient-derived fibroblasts, lymphoblast cells, and minigene splice assays. One antisense oligonucleotide targeting the exonic splice enhancer region significantly induced pseudoexon skipping and increased the expression of normal mRNA. It also rescued FKTN protein production in lymphoblast cells and restored functional O-mannosyl glycosylation of alpha-dystroglycan in patient-derived myotubes. Based on our results, ASO-based splicing correction should be investigated further as a potential treatment for patients with FCMD carrying the DIV.

リンク情報
DOI
https://doi.org/10.1093/hmg/ddac286
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/36426838
URL
https://academic.oup.com/hmg/article/32/8/1301/6847082 本文へのリンクあり
ID情報
  • DOI : 10.1093/hmg/ddac286
  • PubMed ID : 36426838

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