Abstract
Translational read-through-inducing drugs (TRIDs) promote read-through of nonsense mutations, placing them in the spotlight of current gene-based therapeutic research. Here, we compare for the first time the relative efficacies of new-generation aminoglycosides NB30, NB54 and the chemical compound PTC124 on retinal toxicity and read-through efficacy of a nonsense mutation in the USH1C gene, which encodes the scaffold protein harmonin. This mutation causes the human Usher syndrome, the most common form of inherited deaf-blindness. We quantify read-through efficacy of the TRIDs in cell culture and show the restoration of harmonin function. We do not observe significant differences in the read-through efficacy of the TRIDs in retinal cultures; however, we show an excellent biocompatibility in retinal cultures with read-through versus toxicity evidently superior for NB54 and PTC124. In addition, in vivo administration of NB54 and PTC124 induced recovery of the full-length harmonin a1 with the same efficacy. The high biocompatibilities combined with the sustained read-through efficacies of these drugs emphasize the potential of NB54 and PTC124 in treating nonsense mutation-based retinal disorders.
Highlights
In-frame nonsense mutations account for $12% of all hereditary disease-causing mutations (Kellermayer, 2006)
All through-inducing drugs (TRIDs) were able to rescue translational read-though of the p.R31X mutation to some degree resulting in full-length harmonin a1 expression with the highest level of read-through achieved by NB54
All administered TRIDs were able to rescue translational read-though of the p.R31X mutation to some degree resulting in full-length harmonin a1 expression with the highest level of read-through achieved by NB54
Summary
In-frame nonsense mutations account for $12% of all hereditary disease-causing mutations (Kellermayer, 2006). A gene-based therapy that targets in-frame nonsense mutations could treat a substantial proportion of patients making the (1) Cell and Matrix Biology, Institute of Zoology, Johannes Gutenberg University of Mainz, Mainz, Germany (2) Edith and Joseph Fischer Enzyme Inhibitors Laboratory, Schulich Faculty of Chemistry, Technion-Israel Institute of Technology, Haifa, Israel yPresent address: Department for Neuropathology, University Clinic Freiburg, Institute of Pathology, Freiburg, Germany zPresent address: Vetsuisse Faculty, Institute of Animal Pathology, Bern, Switzerland xThese authors share last authorship. Recent studies demonstrate translational read-through (TR) to be an attractive alternative to gene therapy for in-frame nonsense mutations (Hainrichson et al, 2008; Keeling & Bedwell, 2011; Linde & Kerem, 2008; Overlack et al, 2011). Various chemicals referred to as translational read-through-inducing drugs (TRIDs) are known to promote TR. Known TRIDs include clinically used aminoglycoside antibiotics like gentamicin and paromomycin, and several recently newly designed aminoglycosides like NB30 and NB54 as well as the unrelated chemical compound PTC124 (Fig 1B; Hainrichson et al, 2008; Keeling & Bedwell, 2011; Linde & Kerem, 2008)
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