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Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors

Nature Genetics volume 17, pages 314317 (1997) | Download Citation

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Abstract

Successful gene therapy approaches will require efficient gene delivery and sustained expression of the transgene in recipients. A variety of methods, ranging from direct DNA delivery to infection with recombinant viruses containing foreign genes, have been developed, but they all have some major limitations that restrict their utility14. We have described a human lentiviral (HIV)–based vector that can transduce non-dividing cells in vitro and deliver genes In vivo5,6. With this vector, expression of transgenes in the brain has been detected for more than six months—the longest period tested so far7. Because lentiviral vectors are pseudotyped with vesicular stomatitis virus G glycoprotein (VSVG; ref. 8), they can transduce a broad range of tissues and cell types. We now describe the ability of lentiviral vectors to introduce genes directly into liver and muscle. Sustained expression of green fluorescent protein (GFP), used as a surrogate for therapeutic protein/can be observed for more than 22 weeks in the liver. Similar long-term expression (more than eight weeks) was observed in transduced muscle. In contrast, little or no GFP could be detected in liver or muscle transduced with the Moloney murine leukaemia virus (M-MLV), a prototypic retroviral based vector. At a minimum, 3–4% of the total liver tissue was transduced by a single injection of 1–3x107 infectious units (I.U.) of recombinant HIV vector. Furthermore, no inflammation or recruitment of lymphocytes could be detected at the site of injection. Animals previously transduced with a lentiviral vector can be efficiently re-infected with lentiviral vectors. Additionally, we show that the requirement for lentiviral accessory proteins to establish efficient transduction in vivo is tissue dependent.

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References

  1. 1.

    The basic science of gene therapy. Science 260, 926–932 (1993).

  2. 2.

    Transfer of genes to humans: early lessons and obstacles to success. Science 270, 404–410 (1995).

  3. 3.

    Gene therapy–promises, pitfalls and prognosis. N. Engl. J. Med. 333, 871–873 (1995).

  4. 4.

    & Gene therapy: promises, problems and prospects. Nature 389, 239–242 (1997).

  5. 5.

    , , , & Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc. Natl. Acad. Sci. USA 93, 11382–11388 (1996).

  6. 6.

    et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272, 263–267 (1996).

  7. 7.

    et al. Highly efficient and sustained gene transfer in adult neurons with a lentiviral vector. J. Virol. 71, 6641–6649 (1997).

  8. 8.

    , , , & Vesicular stomatitis virus G protein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells. Proc. Natl Acad. Sci. USA 90, 8033–8037 (1993).

  9. 9.

    , , & Stable and efficient gene transfer into the retina using a lentiviral vector. Proc. Natl Acad. Sci. USA 94, 10319–10323 (1997).

  10. 10.

    , , & Perforant path damage results in progressive neuronal death and somal atrophy in layer II of entorhinal cortex and functional impairment with increasing post-damage age. J. Neurosci. 14, 6872–6885 (1994).

  11. 11.

    & Systemic delivery of recombinant proteins by genetically modified myoplasts. Science 254, 1507–1509 (1991).

  12. 12.

    et al. Systemic delivery of human growth hormone by injection of genetically engineered myoblasts. Science 254, 1509–1512 (1991).

  13. 13.

    , , & Gene therapy via primary myoblasts: Long-term expression of factor IX protein following transplantation in vivo. Proc Natl. Acad. Sci. USA 89, 10892–10895 (1992).

  14. 14.

    et al. Cellular and humoral immune responses to adenoviral vectors containing factor IX gene: tolerization of factor IX and vector antigens allows for long-term expression. Proc. Natl Acad. Sci. USA 92, 1401–1405 (1995).

  15. 15.

    , , & Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes. Proc. Natl. Acad. Sci. USA 88, 1330–1334 (1991).

  16. 16.

    , & Long-term in vivo expression of retrovirus-mediated gene transfer in mouse fibroblast implants. Proc Natl Acad. Sci. USA 88, 4626–4630 (1991).

  17. 17.

    , & MHC class l-restricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses. Immunity 1, 433–442 (1994).

  18. 18.

    , , & Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses. J. Virol. 69, 2004–2015 (1995).

  19. 19.

    , & Construction and in vitro properties of HIV-1 mutants with deletions in “non-essential” genes. AIDS Res. Hum. Retroviruses 10, 343–350 (1994).

  20. 20.

    et al. The human immunodeficiency virus type 2 Vpr genes is essential for productive infection of human macrophages. Proc Natl Acad. Sci. USA 87, 8080–8084 (1990).

  21. 21.

    et al. The Vpr protein of human immunodeficiency virus type 1 influences nuclear localization of viral nucleic acids in nondividing host cells. Proc Natl. Acad. Sci. USA 91, 7311–7315 (1994).

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  1. Laboratory of Genetics, The Salk Institute, 10010 North Torrey Pines Road, La Jolla, California 92037, USA

    • Tal Kafri
    • , Ulrike Blömer
    • , Daniel A. Peterson
    • , Fred H. Gage
    •  & Inder M. Verma

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Correspondence to Inder M. Verma.

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DOI

https://doi.org/10.1038/ng1197-314