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

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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 - Feb 2008

All Science Journal Classification (ASJC) codes

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

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    Allamand, V., Bidou, L., Arakawa, M., Floquet, C., Shiozuka, M., Paturneau-Jouas, M., Gartioux, C., Butler-Browne, G. S., Mouly, V., Rousset, J. P., 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, 10(2), 217-224. https://doi.org/10.1002/jgm.1140