Assessment of the feasibility of exon 45-55 multiexon skipping for Duchenne muscular dystrophy.
BACKGROUND
The specific skipping of an exon, induced by antisense oligonucleotides (AON) during splicing, has shown to be a promising therapeutic approach for Duchenne muscular dystrophy (DMD) patients. As different mutations require skipping of different exons, this approach is mutation dependent. The skipping of an entire stretch of exons (e.g. exons 45 to 55) has recently been suggested as an approach applicable to larger groups of patients. However, this multiexon skipping approach is technically challenging. The levels of intended multiexon skips are typically low and highly variable, and may be dependent on the order of intron removal. We hypothesized that the splicing order might favor the induction of multiexon 45-55 skipping.


METHODS
We here tested the feasibility of inducing multiexon 45-55 in control and patient muscle cell cultures using various AON cocktails.


RESULTS
In all experiments, the exon 45-55 skip frequencies were minimal and comparable to those observed in untreated cells.


CONCLUSIONS
We conclude that current state of the art does not sufficiently support clinical development of multiexon skipping for DMD.
van Vliet
Laura
L
DMD Genetic Therapy Group, Department of Human Genetics, Leiden University, Medical Center, Postzone S4-P, PO Box 9600, 2300RC, Leiden, The Netherlands. l.van_vliet@lumc.nl
de Winter
Christa L
CL
van Deutekom
Judith C T
JC
van Ommen
Gert-Jan B
GJ
Aartsma-Rus
Annemieke
A
D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D005091:Exons; D015316:Genetic Therapy; D006801:Humans; D018485:Muscle Fibers, Skeletal / Q000473:pathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D032448:Myoblasts, Skeletal / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D016366:Open Reading Frames; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion
D016376:Oligonucleotides, Antisense