Cellular biological study on the treatment of Duchenne muscular dystrophy with nucleic acids

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Duchenne muscular dystrophy (DMD) is a fatal muscle-wasting disease, and its victims usually succumb in their twenties. Many studies, including investigations into gene-replacement therapy, have been conducted in a search for a treatment for DMD, and the most promising treatment to date is rescue of mutant dystrophin mRNA by induction of exon skipping. On the basis of results from the molecular analysis of dystrophin Kobe, we have proposed a treatment for DMD in which antisense oligonucleotides induce exon skipping to edit out-of frame dystrophin mRNA into in-frame, thereby converting severe DMD to a milder form. Here we conducted studies on development of RNA/ENA chimeric oligonucleotide for the treatment of DMD. At first we searched for the best RNA/ENA chimera that has ability to induce skipping of target exon. After repeating trials to induce exon skipping in cultured myocytes, we succeeded to identify the most powerful RNA/ENA chimera for exon skipping. The identified RNA/ENA chimera was transfected to a DMD patient's cultured myocytes and shown to induce dystrophin expression. This treatment strategy was examined for application to mouse treatment. We obtained a knockout mouse that has a deletion of exon 52 of the dystrophin gene. By identifying a proper RNA/ENA chimera, we are going to induce exon 51 skipping in the model mouse.