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Ameliorating pathogenesis by removing an exon containing a missense mutation: a potential exon-skipping therapy for laminopathies

Gene Therapy volume 22, pages 503–515 (2015)Cite this article

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Abstract

Exon skipping, as a therapy to restore a reading frame or switch protein isoforms, is under clinical trial. We hypothesised that removing an in-frame exon containing a mutation could also improve pathogenic phenotypes. Our model is laminopathies: incurable tissue-specific degenerative diseases associated with LMNA mutations. LMNA encodes A-type lamins, that together with B-type lamins, form the nuclear lamina. Lamins contain an alpha-helical central rod domain composed of multiple heptad repeats. Eliminating LMNA exon 3 or 5 removes six heptad repeats, so shortens, but should not otherwise significantly alter, the alpha-helix. Human Lamin A or Lamin C with a deletion corresponding to amino acids encoded by exon 5 (Lamin A/C-Δ5) localised normally in murine lmna-null cells, rescuing both nuclear shape and endogenous Lamin B1/emerin distribution. However, Lamin A carrying pathogenic mutations in exon 3 or 5, or Lamin A/C-Δ3, did not. Furthermore, Lamin A/C-Δ5 was not deleterious to wild-type cells, unlike the other Lamin A mutants including Lamin A/C-Δ3. Thus Lamin A/C-Δ5 function as effectively as wild-type Lamin A/C and better than mutant versions. Antisense oligonucleotides skipped LMNA exon 5 in human cells, demonstrating the possibility of treating certain laminopathies with this approach. This proof-of-concept is the first to report the therapeutic potential of exon skipping for diseases arising from missense mutations.

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Acknowledgements

We thank Matthew Wood and his laboratory members for their advice. L302P human patient fibroblasts were a kind gift from Gisèle Bonne (Institute of Myology, Paris, France). JS and NF were partially funded by OPTISTEM (223098) through EU FP7. PSZ is also supported by The Muscular Dystrophy Campaign, the Medical Research Council (G1100193), Association Franςaise contre les Myopathies and BIODESIGN (262948-2) through EU FP7.

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  1. Randall Division of Cell and Molecular Biophysics, King's College London, London, UK

    J Scharner, N Figeac, J A Ellis & P S Zammit

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  1. J Scharner
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  2. N Figeac
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Correspondence to P S Zammit.

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Scharner, J., Figeac, N., Ellis, J. et al. Ameliorating pathogenesis by removing an exon containing a missense mutation: a potential exon-skipping therapy for laminopathies. Gene Ther 22, 503–515 (2015). https://doi.org/10.1038/gt.2015.8

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