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.

  • Brief Communication
  • Published:

High expression of transgenes mediated by hybrid retroviral vectors in hepatocytes: comparison of promoters from murine retroviruses in vitro and in vivo

Gene Therapy volume 9, pages 303–306 (2002)Cite this article

Abstract

To achieve high transgene expression in the liver, we have compared the reporter gene expression among various murine retroviral long terminal repeats (LTRs) or leader sequences in vitro. Transient reporter gene expression assays revealed the highest gene expression by the polycythemic strain of spleen focus-forming virus (SFFVp) LTR in differentiated hepatocellular carcinoma cell lines, HuH-7 and PLC/PRF/5. However, remarkable difference was not observed among LTRs in other types of human liver tumor cell lines. Essentially the same results were obtained by infecting these cells with a series of retroviral vectors. Repression of transgene expression was observed by the leader sequences from Moloney murine leukemia virus (MoMLV), but not from mouse embryonic stem cell virus (MESV). Strengths of the promoters were further compared in murine hepatocytes in vivo. Although the proportions of genomic integration were almost the same, higher gene expression was observed by the FMEV-type vector, which contained the SFFVp LTR and the MESV leader, in comparison with that by the MoMLV-based vector. Thus, FMEV-type vectors may represent a novel type of vectors for human gene therapy with hepatocytes.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Mulligan R.C. . The basic science of gene therapy Science 1993 260: 926 926

    Article  CAS  PubMed  Google Scholar 

  2. Kay M.A. et al. In vivo gene therapy of hemophilia B: sustained partial correction in factor IX-deficient dogs Science 1993 262: 117 117

    Article  CAS  PubMed  Google Scholar 

  3. Alt M., Caselmann W.H. . Liver-directed gene therapy: molecular tools and current preclinical and clinical studies J Hepatol 1995 23: 746 746

    Article  CAS  PubMed  Google Scholar 

  4. Li M. et al. Comparison of the expression of a mutant dihydrofolate reductase under control of different internal promoters in retroviral vectors Hum Gene Ther 1992 3: 381 381

    Article  CAS  PubMed  Google Scholar 

  5. Rettinger S.D. et al. Liver-directed gene therapy: quantitative evaluation of promoter elements by using in vivo retroviral transduction Proc Natl Acad Sci USA 1994 91: 1460 1460

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Hafenrichetr D.G. et al. Quantitative evaluation of liver-specific promoters from retroviral vectors after in vivo transduction of hepatocytes Blood 1994 84: 3394 3394

    Google Scholar 

  7. Ostertag W. et al. Transforming genes and target cells of murine spleen focus-forming viruses Adv Cancer Res 1987 48: 193 193

    Article  CAS  PubMed  Google Scholar 

  8. Stocking C., Kollek R., Bergholz U., Ostetag W. . Long -terminal repeat sequences impart hematopoietic transformation properties to the myeloproliferative sarcoma virus Proc Natl Acad Sci USA 1985 82: 5746 5746

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Grez M., Akgum E., Hilberg F., Ostertag W. . Embryonic stem cell virus, a recombinant murine retrovirus with expression in embryonic stem cells Proc Natl Acad Sci USA 1990 87: 9202 9202

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Nakabayashi H. et al. Growth of human hepatoma cells lines with differentiated functions in chemically defined medium Cancer Res 1982 42: 3858 3858

    CAS  PubMed  Google Scholar 

  11. Knowles B.B., Howe C.C., Aden D.P. . Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen Science 1980 209: 497 497

    Article  CAS  PubMed  Google Scholar 

  12. Doi I., Mamba M., Sato J. . Establishment and some characteristics of human hepatoma cell lines Jpn J Cancer Res 1975 66: 385 385

    Google Scholar 

  13. Aden D.P. et al. Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line Nature 1979 282: 615 615

    Article  CAS  PubMed  Google Scholar 

  14. Iemura A., Maruiwa M., Yano H., Kojiro M. . A new human cholangiocellular carcinoma cell line (KMC-1) J Hepatol 1992 15: 288 288

    Article  CAS  PubMed  Google Scholar 

  15. Murakami T. et al. Establishment and characterization of a human combined hepatocholangiocarcinoma cell line and its heterologous transplantation in nude mice Hepatology 1987 7: 551 551

    Article  CAS  PubMed  Google Scholar 

  16. Baum C. et al. The potent enhancer activity of the polycytemic strain of spleen focus-forming virus in hematopoietic cells is governed by a binding site for Sp1 in the upstream control region and by a unique enhancer core motif, creating an exclusive target for PEBP/CBF J Virol 1997 71: 6323 6323

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Danno S. et al. Efficient gene transfer by hybrid retroviral vectors to murine spermatogenic cells Hum Gene Ther 1999 10: 1819 1819

    Article  CAS  PubMed  Google Scholar 

  18. Baum C., Eckert H.G., Ostertag W. . Contribution of the retroviral leader to gene transfer and expression in packaging cells and myeloid stem and progenitor cells Zander A, Ostertag W, Afanasiev BV, Grosveld F (eds); Gene Technology Springer-Verlag 1996 pp 219–228

  19. Baum C., Hegewischbecker S., Eckert H.G., Ostertag W. . Novel retroviral vectors for efficient expression of the multidrug resistance (mdr-1) gene in early hematopoietic cells J Virol 1995 69: 7541 7541

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Baum C., Eckert H.G., Stocking C., Ostertag W. . Activity of Friend mink cell focus-forming retrovirus during myelo-erythroid hematopoiesis Exp Hematol 1996 24: 364 364

    CAS  PubMed  Google Scholar 

  21. Tsuji T. et al. Retroviral vector-mediated gene expression in human CD34+CD38- cells expanded in vitro: cis elements of FMEV are superior to those of Mo-MuLV Hum Gene Ther 2000 11: 271 271

    Article  CAS  PubMed  Google Scholar 

  22. Hilberg F., Stocking C., Ostertag W., Grez M. . Functional analysis of a retroviral host-range mutant: altered long terminal repeat sequences allow expression in embryonal carcinoma cells Proc Natl Acad Sci USA 1987 84: 5232 5232

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Kingston R.E., Okayama, Chen C.A., Rose J.K. . Calcium phosphate transfection Ausubel FM (eds); Current Protocols in Molecular Biology John Wiley 1997 pp 9–228

  24. Ludescher C. et al. Detection of activity of P-glycoprotein in human tumor samples using rhodamine 123 Br J Haematol 1992 82: 161 161

    Article  CAS  PubMed  Google Scholar 

  25. Higgins G.M., Anderson R.M. . Experimental pathology of the liver. I. Restoration of the liver of white rat following partial surgical removal Arch Pathol 1931 12: 186 186

    Google Scholar 

  26. Heid C.A., Stevens J., Livak K.J., Williams P.M. . Real-time quantitative PCR Genome Res 1996 6: 986 986

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr H Yano of Kurume University School of Medicine (Kurume, Japan) for providing us KMC-1 and KMCH-1 cells. This work was partly supported by Grants-in-Aid from the ministry of Education, Science, Sports and Culture of Japan.

Author information

Authors and Affiliations

  1. Department of Clinical Molecular Biology, Faculty of Medicine, Kyoto University, Kyoto, Japan

    N Ohnishi, K Itoh, H Higashitsuji, K Yamaguchi, T Tsuji & J Fujita

  2. Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan

    N Ohnishi, Y Itoh & T Okanoue

  3. Department of Hematology/Oncology, Hannover Medical School, Hannover, Germany

    C Baum

Authors
  1. N Ohnishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K Itoh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y Itoh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C Baum
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H Higashitsuji
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K Yamaguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. T Tsuji
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. T Okanoue
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. J Fujita
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ohnishi, N., Itoh, K., Itoh, Y. et al. High expression of transgenes mediated by hybrid retroviral vectors in hepatocytes: comparison of promoters from murine retroviruses in vitro and in vivo. Gene Ther 9, 303–306 (2002). https://doi.org/10.1038/sj.gt.3301655

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301655

Keywords

This article is cited by

Search

Advanced search

Quick links