Combinatorial Utrophin A Activation in Muscle as a Therapeutic Strategy to Treat Duchenne Muscular Dystrophy

Duchenne Muscular Dystrophy (DMD) is an X-linked recessive neuromuscular disorder caused by mutations or deletions in the dystrophin gene. Utrophin up-regulation therapy is among the various therapeutic strategies that are being investigated to treat DMD. In this strategy utrophin, a dystrophin homologue, is up-regulated along the entire length of the sarcolemma to replace the absent dystrophin protein. Previous studies have revealed that utrophin A expression can be controlled by various transcriptional, post-transcriptional and translational mechanisms and pharmacological modulation of these pathways can stimulate its expression in muscle. In the present study we screened several FDA approved and natural pharmacological compounds that can potentially activate utrophin A expression in muscle. We found that AICAR (AMPK activator) and heparin (p38 activator) were most effective in stimulating utrophin A expression in our C2C12 muscle cell system. Next, we analyzed the effect of combining these activators on utrophin A expression in muscle cells and preclinical mdx mouse model of DMD. Our findings revealed that combinatorial treatment of AICAR and heparin instigated an additive effect on utrophin A expression both in C2C12 muscle cells and mdx mice. Further characterization of treated mdx mice revealed that combinatorial treatment of AICAR and heparin caused improvements in the dystrophic phenotype as indicated by decreased central nucleation, decreased fiber size variability and improved sarcolemmal integrity in dystrophic muscle. Together these findings established that combinatorial treatment of AICAR and heparin ameliorates the dystrophic phenotype in mdx mice and may serve as an effective therapeutic strategy for DMD