Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

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

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.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  4. Verma, I.M. & Somia, N. Gene therapy: promises, problems and prospects. Nature 389, 239–242 (1997).

    Article  CAS  PubMed  Google Scholar 

  5. Naldini, L., Blomer, U., Blomer, R., Trono,, D. & Verma, I. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Burns, J.C., Friedman, T., Driever, W., Burrascano, M. & Yii, J.-K. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Miyoshi, H., Takahashi, M., Gage, F.H. & Verma, I.M. Stable and efficient gene transfer into the retina using a lentiviral vector. Proc. Natl Acad. Sci. USA 94, 10319–10323 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Peterson, D.A., Lucidi-Phillipi, C.A., Eagle, K.L. & Gage, F.H. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Barr, E. & Leiden, J.M. Systemic delivery of recombinant proteins by genetically modified myoplasts. Science 254, 1507–1509 (1991).

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  13. Dai, Y., Roman, M., Naviaux, R.K. & Verma, I.M. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Dai, Y. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Palmer, T.D., Rosman, G.J., Osborne, W.R.A. & Miller, A.D. Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes. Proc. Natl. Acad. Sci. USA 88, 1330–1334 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Scharfmann, R., Axelrod, J.H. & Verma, I.M. Long-term in vivo expression of retrovirus-mediated gene transfer in mouse fibroblast implants. Proc Natl Acad. Sci. USA 88, 4626–4630 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Yang, Y., Ertl, J. & Wilson, J.M. 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).

    Article  CAS  PubMed  Google Scholar 

  18. Yang, Y., Li, Q., Ertl, H. & Wilson, J. Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses. J. Virol. 69, 2004–2015 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Gibbs, J.S., Regier, D.A. & Desrosiers, R.C. Construction and in vitro properties of HIV-1 mutants with deletions in “non-essential” genes. AIDS Res. Hum. Retroviruses 10, 343–350 (1994).

    Article  CAS  PubMed  Google Scholar 

  20. Hattori, N. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Heinzinger, N.K. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Inder M. Verma.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kafri, T., Blömer, U., Peterson, D. et al. Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors. Nat Genet 17, 314–317 (1997). https://doi.org/10.1038/ng1197-314

Download citation

  • Received:

  • Accepted:

  • Issue Date:

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

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing