Antisense Oligonucleotide-based Splice Correction for USH2A-associated Retinal Degeneration Caused by a Frequent Deep-intronic Mutation

Radulfus Wn Slijkerman , Christel Vaché (1, 2) , Margo Dona , Gema García-García (1) , Mireille Claustres (1, 2) , Lisette Hetterschijt , Theo Peters , Bas Hartel , Ronald Je Pennings , José del R. Millán (3) , Elena Aller (4) , Alejandro Garanto , Rob Wj Collin , Hannie Kremer , Anne-Francoise Roux (1, 2) , Erwin van Wijk

1 LGMR - Laboratoire de génétique des maladies rares. Pathologie moleculaire, etudes fonctionnelles et banque de données génétiques
2 CHRU Montpellier - Centre Hospitalier Régional Universitaire [Montpellier]
3 CNM - Centro Nacional de Microelectronica [Spain]
4 Genetics

Usher syndrome (USH) is the most common cause of combined deaf-blindness in man. The hearing loss can be partly compensated by providing patients with hearing aids or cochlear implants, but the loss of vision is currently untreatable. In general, mutations in the USH2A gene are the most frequent cause of USH explaining up to 50% of all patients worldwide. The first deep-intronic mutation in the USH2A gene (c.7595-2144A>G) was reported in 2012, leading to the insertion of a pseudoexon (PE40) into the mature USH2A transcript. When translated, this PE40-containing transcript is predicted to result in a truncated non-functional USH2A protein. In this study, we explored the potential of antisense oligonucleotides (AONs) to prevent aberrant splicing of USH2A pre-mRNA as a consequence of the c.7595-2144A>G mutation. Engineered 2'-O-methylphosphorothioate AONs targeting the PE40 splice acceptor site and/or exonic splice enhancer regions displayed significant splice correction potential in both patient derived fibroblasts and a minigene splice assay for USH2A c.7595-2144A>G, whereas a non-binding sense oligonucleotide had no effect on splicing. Altogether, AON-based splice correction could be a promising approach for the development of a future treatment for USH2A-associated retinitis pigmentosa caused by the deep-intronic c.7595-2144A>G mutation.

Domaines: Sciences du Vivant [q-bio]

Radulfus Wn Slijkerman, Christel Vaché, Margo Dona, Gema García-García, Mireille Claustres, et al.. Antisense Oligonucleotide-based Splice Correction for USH2A-associated Retinal Degeneration Caused by a Frequent Deep-intronic Mutation. Molecular Therapy - Nucleic Acids, 2016, 5, pp.e381. ⟨10.1038/mtna.2016.89⟩. ⟨hal-02436494⟩