Drug-induced readthrough of premature stop codons leads to the stabilization of laminin α2 chain mRNA in CMD myotubes

Valérie Allamand, Laure Bidou, Masayuki Arakawa, Célia Floquet, Masataka Shiozuka, Marion Paturneau-Jouas, Corine Gartioux, Gillian S. Butler-Browne, Vincent Mouly, Jean Pierre Rousset, Ryoichi Matsuda, Daishiro Ikeda, Pascale Guicheney

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Background: The most common form of congenital muscular dystrophy is caused by a deficiency in the α2 chain of laminin-211, a protein of the extracellular matrix. A wide variety of mutations, including 20 to 30% of nonsense mutations, have been identified in the corresponding gene, LAMA2. A promising approach for the treatment of genetic disorders due to premature termination codons (PTCs) is the use of drugs to force stop codon readthrough. Methods: Here, we analyzed the effects of two compounds on a PTC in the LAMA2 gene that targets the mRNA to nonsense-mediated RNA decay, in vitro using a dual reporter assay, as well as ex vivo in patient-derived myotubes. Results: We first showed that both gentamicin and negamycin promote significant readthrough of this PTC. We then demonstrated that the mutant mRNAs were strongly stabilized in patient-derived myotubes after administration of negamycin, but not gentamicin. Nevertheless, neither treatment allowed re-expression of the laminin α2-chain protein, pointing to problems that may have arisen at the translational or post-translational levels. Conclusions: Taken together, our results emphasize that achievement of a clinical benefit upon treatment with novel readthrough-inducing agents would require several favourable conditions including PTC nucleotide context, intrinsic and induced stability of mRNA and correct synthesis of a full-length active protein.

Original languageEnglish
Pages (from-to)217-224
Number of pages8
JournalJournal of Gene Medicine
Volume10
Issue number2
DOIs
Publication statusPublished - Jan 1 2008

Fingerprint

Nonsense Codon
Skeletal Muscle Fibers
Laminin
Messenger RNA
Pharmaceutical Preparations
RNA Stability
Gentamicins
Inborn Genetic Diseases
Muscular Dystrophies
Terminator Codon
Extracellular Matrix Proteins
Genes
Proteins
Therapeutics
Nucleotides
Mutation
negamycin

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Drug Discovery
  • Genetics(clinical)

Cite this

Drug-induced readthrough of premature stop codons leads to the stabilization of laminin α2 chain mRNA in CMD myotubes. / Allamand, Valérie; Bidou, Laure; Arakawa, Masayuki; Floquet, Célia; Shiozuka, Masataka; Paturneau-Jouas, Marion; Gartioux, Corine; Butler-Browne, Gillian S.; Mouly, Vincent; Rousset, Jean Pierre; Matsuda, Ryoichi; Ikeda, Daishiro; Guicheney, Pascale.

In: Journal of Gene Medicine, Vol. 10, No. 2, 01.01.2008, p. 217-224.

Research output: Contribution to journalArticle

Allamand, V, Bidou, L, Arakawa, M, Floquet, C, Shiozuka, M, Paturneau-Jouas, M, Gartioux, C, Butler-Browne, GS, Mouly, V, Rousset, JP, Matsuda, R, Ikeda, D & Guicheney, P 2008, 'Drug-induced readthrough of premature stop codons leads to the stabilization of laminin α2 chain mRNA in CMD myotubes', Journal of Gene Medicine, vol. 10, no. 2, pp. 217-224. https://doi.org/10.1002/jgm.1140
Allamand, Valérie ; Bidou, Laure ; Arakawa, Masayuki ; Floquet, Célia ; Shiozuka, Masataka ; Paturneau-Jouas, Marion ; Gartioux, Corine ; Butler-Browne, Gillian S. ; Mouly, Vincent ; Rousset, Jean Pierre ; Matsuda, Ryoichi ; Ikeda, Daishiro ; Guicheney, Pascale. / Drug-induced readthrough of premature stop codons leads to the stabilization of laminin α2 chain mRNA in CMD myotubes. In: Journal of Gene Medicine. 2008 ; Vol. 10, No. 2. pp. 217-224.
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AU - Bidou, Laure

AU - Arakawa, Masayuki

AU - Floquet, Célia

AU - Shiozuka, Masataka

AU - Paturneau-Jouas, Marion

AU - Gartioux, Corine

AU - Butler-Browne, Gillian S.

AU - Mouly, Vincent

AU - Rousset, Jean Pierre

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