Identification of drug modifiers for RYR1-related myopathy using a multi-species discovery pipeline
Jonathan R Volpatti,
Yukari Endo,
Jessica Knox,
Linda Groom,
Stephanie Brennan,
Ramil Noche,
William J Zuercher,
Peter Roy,
Robert T Dirksen,
James J Dowling
Affiliations
Jonathan R Volpatti
Program for Genetics and Genome Biology, Hospital for Sick Children, Toronto, Canada; Department of Molecular Genetics, University of Toronto, Toronto, Canada
Yukari Endo
Program for Genetics and Genome Biology, Hospital for Sick Children, Toronto, Canada
Jessica Knox
Department of Molecular Genetics, University of Toronto, Toronto, Canada; Department of Pharmacology and Toxicology, University of Toronto, Toronto, Canada
Linda Groom
Department of Pharmacology, University of Rochester, Rochester, United States
Stephanie Brennan
Program for Genetics and Genome Biology, Hospital for Sick Children, Toronto, Canada; Department of Molecular Genetics, University of Toronto, Toronto, Canada
Ramil Noche
Program for Genetics and Genome Biology, Hospital for Sick Children, Toronto, Canada
William J Zuercher
UNC Eshelman School of Pharmacy, SGC Center for Chemical Biology, University of North Carolina, Chapel Hill, United States
Peter Roy
Department of Molecular Genetics, University of Toronto, Toronto, Canada; Department of Pharmacology and Toxicology, University of Toronto, Toronto, Canada
Program for Genetics and Genome Biology, Hospital for Sick Children, Toronto, Canada; Department of Molecular Genetics, University of Toronto, Toronto, Canada
Ryanodine receptor type I-related myopathies (RYR1-RMs) are a common group of childhood muscle diseases associated with severe disabilities and early mortality for which there are no available treatments. The goal of this study is to identify new therapeutic targets for RYR1-RMs. To accomplish this, we developed a discovery pipeline using nematode, zebrafish, and mammalian cell models. We first performed large-scale drug screens in C. elegans which uncovered 74 hits. Targeted testing in zebrafish yielded positive results for two p38 inhibitors. Using mouse myotubes, we found that either pharmacological inhibition or siRNA silencing of p38 impaired caffeine-induced Ca2+ release from wild type cells while promoting intracellular Ca2+ release in Ryr1 knockout cells. Lastly, we demonstrated that p38 inhibition blunts the aberrant temperature-dependent increase in resting Ca2+ in myotubes from an RYR1-RM mouse model. This unique platform for RYR1-RM therapy development is potentially applicable to a broad range of neuromuscular disorders.
