title: Dantrolene enhances antisense-mediated exon skipping in human and mouse models of Duchenne muscular dystrophy.
abstract: Duchenne muscular dystrophy (DMD) causes profound and progressive muscle weakness and loss, resulting in early death. DMD is usually caused by frameshifting deletions in the gene DMD, which leads to absence of dystrophin protein. Dystrophin binds to F-actin and components of the dystrophin-associated glycoprotein complex and protects the sarcolemma from contraction-induced injury. Antisense oligonucleotide-mediated exon skipping is a promising therapeutic approach aimed at restoring the DMD reading frame and allowing expression of an intact dystrophin glycoprotein complex. To date, low levels of dystrophin protein have been produced in humans by this method. We performed a small-molecule screen to identify existing drugs that enhance antisense-directed exon skipping. We found that dantrolene, currently used to treat malignant hyperthermia, potentiates antisense oligomer-guided exon skipping to increase exon skipping to restore the mRNA reading frame, the sarcolemmal dystrophin protein, and the dystrophin glycoprotein complex in skeletal muscles of mdx mice when delivered intramuscularly or intravenously. Further, dantrolene synergized with multiple weekly injections of antisense to increase muscle strength and reduce serum creatine kinase in mdx mice. Dantrolene similarly promoted antisense-mediated exon skipping in reprogrammed myotubes from DMD patients. Ryanodine and Rycal S107, which, like dantrolene, targets the ryanodine receptor, also promoted antisense-driven exon skipping, implicating the ryanodine receptor as the critical molecular target.
authors.affiliation: Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
authors.forename: Genevieve C
authors.initials: GC
authors.lastname: Kendall
authors.forename: Ekaterina I
authors.initials: EI
authors.lastname: Mokhonova
authors.forename: Miriana
authors.initials: M
authors.lastname: Moran
authors.forename: Natalia E
authors.initials: NE
authors.lastname: Sejbuk
authors.forename: Derek W
authors.initials: DW
authors.lastname: Wang
authors.forename: Oscar
authors.initials: O
authors.lastname: Silva
authors.forename: Richard T
authors.initials: RT
authors.lastname: Wang
authors.forename: Leonel
authors.initials: L
authors.lastname: Martinez
authors.forename: Qi L
authors.initials: QL
authors.lastname: Lu
authors.forename: Robert
authors.initials: R
authors.lastname: Damoiseaux
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Spencer
authors.forename: Stanley F
authors.initials: SF
authors.lastname: Nelson
authors.forename: M Carrie
authors.initials: MC
authors.lastname: Miceli
mesh_terms: D000818:Animals; D002460:Cell Line; D003620:Dantrolene / Q000008:administration & dosage / Q000494:pharmacology*; D004195:Disease Models, Animal; D004357:Drug Synergism; D016189:Dystrophin / Q000378:metabolism; D005091:Exons / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007273:Injections, Intramuscular; D051379:Mice; D008810:Mice, Inbred C57BL; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009137:Muscular Dystrophy, Animal / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology*; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D012508:Sarcolemma / Q000187:drug effects / Q000378:metabolism / Q000473:pathology
chemical_list: D016189:Dystrophin; D016376:Oligonucleotides, Antisense; D019837:Ryanodine Receptor Calcium Release Channel; D003620:Dantrolene