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PTC124 targets genetic disorders caused by nonsense mutations

Abstract

Nonsense mutations promote premature translational termination and cause anywhere from 5–70% of the individual cases of most inherited diseases1. Studies on nonsense-mediated cystic fibrosis have indicated that boosting specific protein synthesis from <1% to as little as 5% of normal levels may greatly reduce the severity or eliminate the principal manifestations of disease2,3. To address the need for a drug capable of suppressing premature termination, we identified PTC124—a new chemical entity that selectively induces ribosomal readthrough of premature but not normal termination codons. PTC124 activity, optimized using nonsense-containing reporters, promoted dystrophin production in primary muscle cells from humans and mdx mice expressing dystrophin nonsense alleles, and rescued striated muscle function in mdx mice within 2–8 weeks of drug exposure. PTC124 was well tolerated in animals at plasma exposures substantially in excess of those required for nonsense suppression. The selectivity of PTC124 for premature termination codons, its well characterized activity profile, oral bioavailability and pharmacological properties indicate that this drug may have broad clinical potential for the treatment of a large group of genetic disorders with limited or no therapeutic options.

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Figure 1: PTC124 suppresses premature nonsense codons.
Figure 2: Full-length dystrophin is produced in PTC124-treated cultured myotubes.
Figure 3: Rescue of the dystrophic phenotype in muscles of the mdx mouse.
Figure 4: PTC124 activity is selective for readthrough of premature translation termination codons.

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References

  1. Mendell, J. T. & Dietz, H. C. When the message goes awry: disease-producing mutations that influence mRNA content and performance. Cell 107, 411–414 (2001)

    Article  CAS  Google Scholar 

  2. Kerem, E. Pharmacologic therapy for stop mutations: how much CFTR activity is enough? Curr. Opin. Pulm. Med. 10, 547–552 (2004)

    Article  CAS  Google Scholar 

  3. Ramalho, A. S. et al. Five percent of normal cystic fibrosis transmembrane conductance regulator mRNA ameliorates the severity of pulmonary disease in cystic fibrosis. Am. J. Respir. Cell Mol. Biol. 27, 619–627 (2002)

    Article  CAS  Google Scholar 

  4. Maquat, L. E. Nonsense-mediated mRNA decay: splicing, translation and mRNP dynamics. Nature Rev. Mol. Cell Biol. 5, 89–99 (2004)

    Article  CAS  Google Scholar 

  5. Amrani, N. Sachs M. S. & Jacobson, A. Early nonsense: mRNA decay solves a translational problem. Nature Rev. Mol. Cell Biol. 7, 415–425 (2006)

    Article  CAS  Google Scholar 

  6. Welch, E. M., Wang, W. & Peltz, S. W. in Translational Control of Gene Expression (eds Sonenberg, N., Hershey, J. W. B. & Mathews, M. B.) 467–486 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2000)

    Google Scholar 

  7. Maderazo, A. B., He, F., Mangus, D. A. & Jacobson, A. Upf1p control of nonsense mRNA translation is regulated by Nmd2p and Upf3p. Mol. Cell. Biol. 20, 4591–4603 (2000)

    Article  CAS  Google Scholar 

  8. Weng, Y., Czaplinski, K. & Peltz, S. W. Genetic and biochemical characterization of mutations in the ATPase and helicase regions of the Upf1 protein. Mol. Cell. Biol. 16, 5477–5490 (1996)

    Article  CAS  Google Scholar 

  9. Weng, Y., Czaplinski, K. & Peltz, S. W. Identification and characterization of mutations in the UPF1 gene that affect nonsense suppression and the formation of the Upf protein complex but not mRNA turnover. Mol. Cell. Biol. 16, 5491–5506 (1996)

    Article  CAS  Google Scholar 

  10. Wang, W., Czaplinski, K., Rao, Y. & Peltz, S. W. The role of Upf proteins in modulating the translation read-through of nonsense-containing transcripts. EMBO J. 20, 880–890 (2001)

    Article  CAS  Google Scholar 

  11. Manuvakhova, M., Keeling, K. & Bedwell, D. M. Aminoglycoside antibiotics mediate context-dependent suppression of termination codons in a mammalian translation system. RNA 6, 1044–1055 (2000)

    Article  CAS  Google Scholar 

  12. Politano, L. et al. Gentamicin administration in Duchenne patients with premature stop codon. Preliminary results. Acta Myol. 22, 15–21 (2003)

    CAS  PubMed  Google Scholar 

  13. Clancy, J. P. et al. Evidence that systemic gentamicin suppresses premature stop mutations in patients with cystic fibrosis. Am. J. Respir. Crit. Care Med. 163, 1683–1692 (2001)

    Article  CAS  Google Scholar 

  14. Wilschanski, M. et al. Gentamicin-induced correction of CFTR function in patients with cystic fibrosis and CFTR stop mutations. N. Engl. J. Med. 349, 1433–1441 (2003)

    Article  CAS  Google Scholar 

  15. Wagner, K. R. et al. Gentamicin treatment of Duchenne and Becker muscular dystrophy due to nonsense mutations. Ann. Neurol. 49, 706–711 (2001)

    Article  CAS  Google Scholar 

  16. Barton-Davis, E. R., Cordier, L., Shoturma, D. I., Leland, S. E. & Sweeney, H. L. Aminoglycoside antibiotics restore dystrophin function to skeletal muscles of mdx mice. J. Clin. Invest. 104, 375–381 (1999)

    Article  CAS  Google Scholar 

  17. Bonetti, B., Fu, L., Moon, J. & Bedwell, D. M. The efficiency of translation termination is determined by a synergistic interplay between upstream and downstream sequences in Saccharomyces cerevisiae. J. Mol. Biol. 251, 334–345 (1995)

    Article  CAS  Google Scholar 

  18. McCaughan, K. K., Brown, C. M., Dalphin, M. E., Berry, M. J. & Tate, W. P. Translational termination efficiency in mammals is influenced by the base following the stop codon. Proc. Natl Acad. Sci. USA 92, 5431–5435 (1995)

    Article  ADS  CAS  Google Scholar 

  19. Howard, M. T. et al. Sequence specificity of aminoglycoside-induced stop condon readthrough: potential implications for treatment of Duchenne muscular dystrophy. Ann. Neurol. 48, 164–169 (2000)

    Article  CAS  Google Scholar 

  20. Petrof, B. J., Shrager, J. B., Stedman, H. H., Kelly, A. M. & Sweeney, H. L. Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc. Natl Acad. Sci. USA 90, 3710–3714 (1993)

    Article  ADS  CAS  Google Scholar 

  21. Jacobson, A. & Peltz, S. W. Interrelationships of the pathways of mRNA decay and translation in eukaryotic cells. Annu. Rev. Biochem. 65, 693–739 (1996)

    Article  CAS  Google Scholar 

  22. Mendell, J. T., Sharifi, N. A., Meyers, J. L., Martinez-Murillo, F. & Dietz, H. C. Nonsense surveillance regulates expression of diverse classes of mammalian transcripts and mutes genomic noise. Nature Genet. 36, 1073–1078 (2004)

    Article  CAS  Google Scholar 

  23. Amrani, N. et al. A faux 3′-UTR promotes aberrant termination and triggers nonsense-mediated mRNA decay. Nature 432, 112–118 (2004)

    Article  ADS  CAS  Google Scholar 

  24. Hirawat, S. et al. Safety, tolerability, and pharmacokinetics of PTC124, a nonaminoglycoside nonsense mutation suppressor, following single- and multiple-dose administration to healthy male and female adult volunteers. J. Clin. Pharm. 47, 430–444 (2007)

    Article  CAS  Google Scholar 

  25. He, F. et al. Genome-wide analysis of mRNAs regulated by the nonsense-mediated and 5′ to 3′ mRNA decay pathways in yeast. Mol. Cell 12, 1439–1452 (2003)

    Article  CAS  Google Scholar 

  26. Pruitt, K. D. & Maglott, D. R. RefSeq and LocusLink: NCBI gene-centered resources. Nucleic Acids Res. 29, 137–140 (2001)

    Article  CAS  Google Scholar 

  27. Frischmeyer, P. A. et al. An mRNA surveillance mechanism that eliminates transcripts lacking termination codons. Science 295, 2258–2261 (2002)

    Article  ADS  CAS  Google Scholar 

  28. van Hoof, H. A., Frischmeyer, P. A., Dietz, H. C. & Parker, R. Exosome-mediated recognition and degradation of mRNAs lacking a termination codon. Science 295, 2262–2264 (2002)

    Article  ADS  CAS  Google Scholar 

  29. Sambrook, J., Fritsch, E. & Maniatis, T. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor. (1989)

    Google Scholar 

  30. Grentzmann, G., Ingram, J. A., Kelly, P. J., Gesteland, R. F. & Atkins, J. F. A dual-luciferase reporter system for studying recoding signals. RNA 4, 479–486 (1998)

    Article  CAS  Google Scholar 

  31. Van Der Velden, V., Kaminski, A., Jackson, R. J. & Belsham, G. J. Defective point mutants of the encephalomyocarditis virus internal ribosome entry site can be complemented in trans. Virology 214, 82–90 (1995)

    Article  CAS  Google Scholar 

  32. Jang, S. K. et al. A segment of the 5′-nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation. J. Virol. 62, 2636–2643 (1988)

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Neville, C., Rosenthal, N., McGrew, M., Bogdanova, N. & Hauschka, S. in Methods in Cell Biology 52. (eds Sweeney H. L. and Emerson, C.) 85–116 (Academic Press, San Diego, 1998)

    Google Scholar 

  34. Sweeney, H. L. & Feng, H. Structure–function analysis of cytoskeletal/contractile proteins in avian myotubes. Methods Cell Biol. 52, 275–282 (1997)

    Article  CAS  Google Scholar 

  35. O'Farrell, P. H. High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021 (1975)

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by an STTR grant to A.J. from the NIH and grants from the Muscular Dystrophy Association (USA) and Parent Project Muscular Dystrophy (USA) to H.L.S. We thank L. Cao and T. Komatsu for helpful discussions, G. Elfring for statistical support, D. Minn, X. Kang and S. Gothe for database mining and informatics expertise, and N. Garneau, S. I. Huq, and A. Bhattacharya for technical expertise. We thank K. Donnelly, C. Hirawat and F. P. Nigel for their effort, support and enthusiasm for the project. L. Gold, D. Goeddel and the late R. Swanson provided advice, encouragement and support at the onset of this project, which is gratefully acknowledged. We are grateful to the patients and their families and doctors for their participation in the clinical trial that generated the muscle biopsies and for their commitment during the development of PTC124.

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All authors have competing financial interests except E. Barton, P. Spatrick, F. He, M. Kawana and H. Feng.

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This file contains Supplementary Results, Supplementary Tables 1- 7, Supplementary Figures 1-7 with Legends and additional references. (PDF 649 kb)

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Welch, E., Barton, E., Zhuo, J. et al. PTC124 targets genetic disorders caused by nonsense mutations. Nature 447, 87–91 (2007). https://doi.org/10.1038/nature05756

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