Gene Therapy via Trans-Splicing for LMNA-Related Congenital Muscular Dystrophy

• Main Authors: Feriel Azibani (Author), Astrid Brull (Author), Ludovic Arandel (Author), Maud Beuvin (Author), Isabelle Nelson (Author), Arnaud Jollet (Author), Esma Ziat (Author), Bernard Prudhon (Author), Sofia Benkhelifa-Ziyyat (Author), Marc Bitoun (Author), Stéphanie Lorain (Author), Gisèle Bonne (Author), Anne T. Bertrand (Author)
• Format: Book
• Published: Elsevier, 2018-03-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

We assessed the potential of Lmna-mRNA repair by spliceosome-mediated RNA trans-splicing as a therapeutic approach for LMNA-related congenital muscular dystrophy. This gene therapy strategy leads to reduction of mutated transcript expression for the benefit of corresponding wild-type (WT) transcripts. We developed 5'-RNA pre-trans-splicing molecules containing the first five exons of Lmna and targeting intron 5 of Lmna pre-mRNA. Among nine pre-trans-splicing molecules, differing in the targeted sequence in intron 5 and tested in C2C12 myoblasts, three induced trans-splicing events on endogenous Lmna mRNA and confirmed at protein level. Further analyses performed in primary myotubes derived from an LMNA-related congenital muscular dystrophy (L-CMD) mouse model led to a partial rescue of the mutant phenotype. Finally, we tested this approach in vivo using adeno-associated virus (AAV) delivery in newborn mice and showed that trans-splicing events occurred in WT mice 50 days after AAV delivery, although at a low rate. Altogether, while these results provide the first evidence for reprogramming LMNA mRNA in vitro, strategies to improve the rate of trans-splicing events still need to be developed for efficient application of this therapeutic approach in vivo.

Item Description

2162-2531
10.1016/j.omtn.2017.12.012