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Adeno-associated virus-mediated gene transfer of the heart/muscle adenine nucleotide translocator (ANT) in mouse

Gene Therapy volume 12, pages 570–578 (2005)Cite this article

Abstract

Mitochondrial myopathy, associated with muscle weakness and progressive external ophthalmoplegia, is caused by mutations in mitochondria oxidative phosphorylation genes including the heart–muscle isoform of the mitochondrial adenine nucleotide translocator (ANT1). To develop therapies for mitochondrial disease, we have prepared a recombinant adeno-associated viral vector (rAAV) carrying the mouse Ant1 cDNA. This vector has been used to transduce muscle cells and muscle from Ant1 mutant mice, which manifest mitochondrial myopathy. AAV-ANT1 transduction resulted in long-term, stable expression of the Ant1 transgene in muscle precursor cells as well as differentiated muscle fibers. The transgene ANT1 protein was targeted to the mitochondrion, was inserted into the mitochondrial inner membrane, formed a functional ADP/ATP carrier, increased the mitochondrial export of ATP and reversed the histopathological changes associated with the mitochondrial myopathy. Thus, AAV transduction has the potential of providing symptomatic relief for the ophthalmoplegia and ptosis resulting from paralysis of the extraocular eye muscles cause by mutations in the Ant1 gene.

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References

  1. Morgan-Hughes JA . Mitochondrial diseases of muscle. Curr Opin Neurol 1994; 7: 457–462.

    Article  CAS  PubMed  Google Scholar 

  2. Wallace DC . Mitochondrial diseases in man and mouse. Science 1999; 283: 1482–1488.

    Article  CAS  PubMed  Google Scholar 

  3. Zeviani M et al. An autosomal dominant disorder with multiple deletions of mitochondrial DNA starting at the D-loop region. Nature 1989; 339: 309–311.

    Article  CAS  PubMed  Google Scholar 

  4. Kaukonen J et al. Role of adenine nucleotide translocator 1 in mtDNA maintenance. Science 2000; 289: 782–785.

    Article  CAS  PubMed  Google Scholar 

  5. Pebay-Peyroula E et al. Structure of mitochondrial ADP/ATP carrier in complex with carboxyatractyloside. Nature 2003; 426: 39–44.

    Article  CAS  PubMed  Google Scholar 

  6. Gropp T et al. Kinetics of electrogenic transport by the ADP/ATP carrier. Biophys J 1999; 77: 714–726.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Kim JS, He L, Lemasters JJ . Mitochondrial permeability transition: a common pathway to necrosis and apoptosis. Biochem Biophys Res Commun 2003; 304: 463–470.

    Article  CAS  PubMed  Google Scholar 

  8. Levy SE, Chen YS, Graham BH, Wallace DC . Expression and sequence analysis of the mouse adenine nucleotide translocase 1 and 2 genes. Gene 2000; 254: 57–66.

    Article  CAS  PubMed  Google Scholar 

  9. Graham BH et al. A mouse model for mitochondrial myopathy and cardiomyopathy resulting from a deficiency in the heart/muscle isoform of the adenine nucleotide translocator. Nat Genet 1997; 16: 226–234.

    Article  CAS  PubMed  Google Scholar 

  10. Smythe GM, Hodgetts SI, Grounds MD . Immunobiology and the future of myoblast transfer therapy. Mol Ther 2000; 1: 304–313.

    Article  CAS  PubMed  Google Scholar 

  11. Li S, Benninger M . Applications of muscle electroporation gene therapy. Curr Gene Ther 2002; 2: 101–105.

    Article  CAS  PubMed  Google Scholar 

  12. Watchko J et al. Adeno-associated virus vector-mediated minidystrophin gene therapy improves dystrophic muscle contractile function in mdx mice. Hum Gene Ther 2002; 13: 1451–1460.

    Article  CAS  PubMed  Google Scholar 

  13. Pruchnic R et al. The use of adeno-associated virus to circumvent the maturation-dependent viral transduction of muscle fibers. Hum Gene Ther 2000; 11: 521–536.

    Article  CAS  PubMed  Google Scholar 

  14. Hamilton H, Gomos J, Berns KI, Falck-Pedersen E . Adeno-associated virus site-specific integration and AAVS1 disruption. J Virol 2004; 78: 7874–7882.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Owen RI et al. Recombinant adeno-associated virus vector-based gene transfer for defects in oxidative metabolism. Hum Gene Ther 2000; 11: 2067–2078.

    Article  CAS  PubMed  Google Scholar 

  16. Seo BB et al. Use of the NADH-quinone oxidoreductase (NDI1) gene of Saccharomyces cerevisiae as a possible cure for complex I defects in human cells. J Biol Chem 2000; 275: 37774–37778.

    Article  CAS  PubMed  Google Scholar 

  17. Xiao X, Li J, Samulski RJ . Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J Virol 1998; 72: 2224–2232.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Snyder RO et al. Efficient and stable adeno-associated virus-mediated transduction in the skeletal muscle of adult immunocompetent mice. Hum Gene Ther 1997; 8: 1891–1900.

    Article  CAS  PubMed  Google Scholar 

  19. Snow MH . The effects of aging on satellite cells in skeletal muscles of mice and rats. Cell Tissue Res 1977; 185: 399–408.

    Article  CAS  PubMed  Google Scholar 

  20. Pette D, Staron RS . Myosin isoforms, muscle fiber types, and transitions. Microsc Res Technol 2000; 50: 500–509.

    Article  CAS  Google Scholar 

  21. Black JT, Judge D, Demers L, Gordon S . Ragged-red fibers. A biochemical and morphological study. J Neurol Sci 1975; 26: 479–488.

    Article  CAS  PubMed  Google Scholar 

  22. Hamalainen N, Pette D . Patterns of myosin isoforms in mammalian skeletal muscle fibres. Microsc Res Technol 1995; 30: 381–389.

    Article  CAS  Google Scholar 

  23. Abadie J et al. Recombinant adeno-associated virus type 2 mediates highly efficient gene transfer in regenerating rat skeletal muscle. Gene Therapy 2002; 9: 1037–1043.

    Article  CAS  PubMed  Google Scholar 

  24. McFarland DC et al. Variation in fibroblast growth factor response and heparan sulfate proteoglycan production in satellite cell populations. Comp Biochem Physiol C 2003; 134: 341–351.

    Google Scholar 

  25. Kokoszka JE et al. The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore. Nature 2004; 427: 461–465.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Fagiolari G et al. Lack of apoptosis in patients with progressive external ophthalmoplegia and mutated adenine nucleotide translocator-1 gene. Muscle Nerve 2002; 26: 265–269.

    Article  PubMed  Google Scholar 

  27. Bauer MK, Schubert A, Rocks O, Grimm S . Adenine nucleotide translocase-1, a component of the permeability transition pore, can dominantly induce apoptosis. J Cell Biol 1999; 147: 1493–1502.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Marti R, Nishigaki Y, Vila MR, Hirano M . Alteration of nucleotide metabolism: a new mechanism for mitochondrial disorders. Clin Chem Lab Med 2003; 41: 845–851.

    Article  CAS  PubMed  Google Scholar 

  29. Luciakova K et al. Repression of the human adenine nucleotide translocase-2 gene in growth-arrested human diploid cells: the role of nuclear factor-1. J Biol Chem 2003; 278: 30624–30633.

    Article  CAS  PubMed  Google Scholar 

  30. Rabinowitz JE et al. Cross-packaging of a single adeno-associated virus (AAV) type 2 vector genome into multiple AAV serotypes enables transduction with broad specificity. J Virol 2002; 76: 791–801.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Shi W, Arnold GS, Bartlett JS . Insertional mutagenesis of the adeno-associated virus type 2 (AAV2) capsid gene and generation of AAV2 vectors targeted to alternative cell-surface receptors. Hum Gene Ther 2001; 12: 1697–1711.

    Article  CAS  PubMed  Google Scholar 

  32. McCarty DM, Monahan PE, Samulski RJ . Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis. Gene Therapy 2001; 8: 1248–1254.

    Article  CAS  PubMed  Google Scholar 

  33. Zolotukhin S et al. A "humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells. J Virol 1996; 70: 4646–4654.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Rando TA, Blau HM . Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy. J Cell Biol 1994; 125: 1275–1287.

    Article  CAS  PubMed  Google Scholar 

  35. Kessler PD et al. Gene delivery to skeletal muscle results in sustained expression and systemic delivery of a therapeutic protein. Proc Natl Acad Sci USA 1996; 93: 14082–14087.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Daly TM et al. Neonatal gene transfer leads to widespread correction of pathology in a murine model of lysosomal storage disease. Proc Natl Acad Sci USA 1999; 96: 2296–2300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Ford T, Graham J, Rickwood D . Iodixanol: a nonionic iso-osmotic centrifugation medium for the formation of self-generated gradients. Anal Biochem 1994; 220: 360–366.

    Article  CAS  PubMed  Google Scholar 

  38. Graham J, Ford T, Rickwood D . The preparation of subcellular organelles from mouse liver in self- generated gradients of iodixanol. Anal Biochem 1994; 220: 367–373.

    Article  CAS  PubMed  Google Scholar 

  39. Chance B, Williams GR . The respiratory chain and oxidative phosphorylation. Adv Enzymol Relat Subj Biochem 1956; 17: 65–134.

    CAS  PubMed  Google Scholar 

  40. Marsh M et al. Rapid analytical and preparative isolation of functional endosomes by free flow electrophoresis. J Cell Biol 1987; 104: 875–886.

    Article  CAS  PubMed  Google Scholar 

  41. Pedersen PL et al. Preparation and characterization of mitochondria and submitochondrial particles of rat liver and liver-derived tissues. Methods Cell Biol 1978; 20: 411–481.

    Article  CAS  PubMed  Google Scholar 

  42. Boulay F, Brandolin G, Lauquin GJ, Vignais PV . Synthesis and properties of fluorescent derivatives of atractyloside as potential probes of the mitochondrial ADP/ATP carrier protein. Anal Biochem 1983; 128: 323–330.

    Article  CAS  PubMed  Google Scholar 

  43. Fiore C et al. Fluorometric detection of ADP/ATP carrier deficiency in human muscle. Clin Chim Acta 2001; 311: 125–135.

    Article  CAS  PubMed  Google Scholar 

  44. Moriyama T, Srere PA . Purification of rat heart and rat liver citrate synthases. Physical, kinetic, and immunological studies. J Biol Chem 1971; 246: 3217–3223.

    CAS  PubMed  Google Scholar 

  45. Manfredi G, Yang L, Gajewski CD, Mattiazzi M . Measurements of ATP in mammalian cells. Methods 2002; 26: 317–326.

    Article  CAS  PubMed  Google Scholar 

  46. Bancroft JDG, Marylin . Theory and Practice of Histological Techniques. WB Saunders Co: Philadelphia, 2002.

    Google Scholar 

  47. Zheng XX, Shoffner JM, Voljavec AS, Wallace DC . Evaluation of procedures for assaying oxidative phosphorylation enzyme activities in mitochondrial myopathy muscle biopsies. Biochim Biophys Acta 1990; 1019: 1–10.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by NIH Grants NS21328, NS41850, AG131154, HL64017 and an Ellison Foundation Senior Investigator Grant awarded to DCW. Support for RJS was provided by an NIH/NHLBI Grant 5P01HL051818-06-10 of the Cystic Fibrosis Foundation.

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Authors and Affiliations

  1. MAMMAG, University of California, Irvine, CA, USA

    A Flierl, P E Coskun & D C Wallace

  2. Knapp Center, University of Chicago, Chicago, IL, USA

    Y Chen

  3. Gene Therapy Center, University of North Carolina, Chapel Hill, NC, USA

    R J Samulski

  4. formerly Center for Mitochondrial Medicine, Emory University,

    A Flierl, P E Coskun & D C Wallace

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  1. A Flierl
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Flierl, A., Chen, Y., Coskun, P. et al. Adeno-associated virus-mediated gene transfer of the heart/muscle adenine nucleotide translocator (ANT) in mouse. Gene Ther 12, 570–578 (2005). https://doi.org/10.1038/sj.gt.3302443

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