Drug-induced readthrough of premature stop codons leads to the stabilization of laminin alpha2 chain mRNA in CMD myotubes. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue The Journal of Gene Medicine Année : 2007

Drug-induced readthrough of premature stop codons leads to the stabilization of laminin alpha2 chain mRNA in CMD myotubes.

Valérie Allamand
Masayuki Arakawa
  • Fonction : Auteur
Masataka Shiozuka
  • Fonction : Auteur
Marion Paturneau-Jouas
  • Fonction : Auteur
Corine Gartioux
Gillian S Butler-Browne
  • Fonction : Auteur
Vincent Mouly
Ryoichi Matsuda
  • Fonction : Auteur
Daishiro Ikeda
  • Fonction : Auteur
Pascale Guicheney
  • Fonction : Auteur

Résumé

BACKGROUND: The most common form of congenital muscular dystrophy is caused by a deficiency in the alpha2 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 alpha2-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. Copyright (c) 2007 John Wiley & Sons, Ltd.

Dates et versions

hal-00198869 , version 1 (18-12-2007)

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Citer

Valérie Allamand, Laure Bidou, Masayuki Arakawa, Célia Floquet, Masataka Shiozuka, et al.. Drug-induced readthrough of premature stop codons leads to the stabilization of laminin alpha2 chain mRNA in CMD myotubes.. The Journal of Gene Medicine, 2007, epub ahead of print. ⟨10.1002/jgm.1140⟩. ⟨hal-00198869⟩
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