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.

  • Original Article
  • Published:

Improved gene transfer selectivity to hepatocarcinoma cells by retrovirus vector displaying single-chain variable fragment antibody against c-Met

Cancer Gene Therapy volume 10, pages 840–849 (2003)Cite this article

Abstract

Engineered retroviruses are widely used vectors for cancer gene therapy approaches. However, the ability to target cells of therapeutic interest while controlling the expression of the transferred genes would improve both the efficiency and the safety of viral vectors. In this study, we investigated the ability of a retroviral amphotropic envelope displaying single-chain variable-fragment (scFv) directed against the c-Met receptor, to target the entry of recombinant retroviruses to human hepatocarcinoma cells. Four single-chain antibody fragments directed against the c-Met receptor were generated and inserted into the viral envelope protein as an N-terminal fusion. The modified envelopes were incorporated into virus particles and one of the chimeric viruses, 3D6-Env, transduced preferentially human hepatoma cells rather than proliferating human hepatocytes. In another construct, the urokinase cleavage site was inserted between the scFv moiety and the envelope. Chimeric scFv-urokinase-Env viruses transduced hepatoma cells with a similar efficiency to that of the control virus and their infectivity in human hepatocytes remained low. These results indicate that amphotropic retroviruses with engineered envelopes to display scFv directed against the c-Met receptor can efficiently and selectively deliver genes into hepatoma cells.

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
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Gagandeep S, Brew R, Green B, et al. Prodrug-activated gene therapy: involvement of an immunological component in the “bystander effect”. Cancer Gene Ther. 1996;3:83–88.

    CAS  Google Scholar 

  2. Valerie K, Hawkins W, Farnsworth J, et al. Substantially improved in vivo radiosensitization of rat glioma with mutant HSV-TK and acyclovir. Cancer Gene Ther. 2001;8:3–83.

    Article  CAS  Google Scholar 

  3. Komatsu K, Suzuki S, Shimosegawa T, Miyazaki JI, Toyota T . Cre-loxP-mediated bax gene activation reduces growth rate and increases sensitivity to chemotherapeutic agents in human gastric cancer cells. Cancer Gene Ther. 2000;7:885–892.

    Article  CAS  Google Scholar 

  4. van der Eb MM, Cramer SJ, Vergouwe Y, et al. Severe hepatic dysfunction after adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene and ganciclovir administration. Gene Therapy. 1998;5:451–458.

    Article  CAS  Google Scholar 

  5. Anderson LM, Krotz S, Weitzman SA, Thimmapaya B . Breast cancer-specific expression of the Candida albicans cytosine deaminase gene using a transcriptional targeting approach. Cancer Gene Ther. 2000;7:845–852.

    Article  CAS  Google Scholar 

  6. Martiniello-Wilks R, Tsatralis T, Russell P, et al. Transcription-targeted gene therapy for androgen-independent prostate cancer. Cancer Gene Ther. 2002;9:443–452.

    Article  CAS  Google Scholar 

  7. Arbuthnot P, Bralet MP, Thomassin H, Danan JL, Brechot C, Ferry N . Hepatoma cell-specific expression of a retrovirally transferred gene is achieved by alpha-fetoprotein but not insulinlike growth factor II regulatory sequences. Hepatology. 1995;22:1788–1796.

    CAS  PubMed  Google Scholar 

  8. Arbuthnot PB, Bralet MP, Le Jossic C, et al. In vitro and in vivo hepatoma cell-specific expression of a gene transferred with an adenoviral vector. Hum Gene Ther. 1996;7:1503–1514.

    Article  CAS  Google Scholar 

  9. Kanai F, Lan KH, Shiratori Y, et al. In vivo gene therapy for alpha-fetoprotein-producing hepatocellular carcinoma by adenovirus-mediated transfer of cytosine deaminase gene. Cancer Res. 1997;57:461–465.

    CAS  PubMed  Google Scholar 

  10. Ohguchi S, Nakatsukasa H, Higashi T, et al. Expression of alpha-fetoprotein and albumin genes in human hepatocellular carcinomas: limitations in the application of the genes for targeting human hepatocellular carcinoma in gene therapy. Hepatology. 1998;27:599–607.

    Article  CAS  Google Scholar 

  11. Mulligan RC . The basic science of gene therapy. Science. 1993;260:926–932.

    Article  CAS  Google Scholar 

  12. Curiel DT . Strategies to adapt adenoviral vectors for targeted delivery. Ann NY Acad Sci. 1999;886:158–171.

    Article  CAS  Google Scholar 

  13. Roe T, Reynolds TC, Yu G, Brown PO . Integration of murine leukemia virus DNA depends on mitosis. EMBO J. 1993;12:2099–2108.

    Article  CAS  Google Scholar 

  14. Ragheb JA, Anderson WF . Uncoupled expression of Moloney murine leukemia virus envelope polypeptides SU and TM: a functional analysis of the role of TM domains in viral entry. J Virol. 1994;68:3207–3219.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Fielding AK, Maurice M, Morling FJ, Cosset FL, Russell SJ . Inverse targeting of retroviral vectors: selective gene transfer in a mixed population of hematopoietic and nonhematopoietic cells. Blood. 1998;91:1802–1809.

    CAS  PubMed  Google Scholar 

  16. Chadwick MP, Morling FJ, Cosset FL, Russell SJ . Modification of retroviral tropism by display of IGF-I. J Mol Biol. 1999;285:485–494.

    Article  CAS  Google Scholar 

  17. Peng KW, Morling FJ, Cosset FL, Murphy G, Russell SJ . A gene delivery system activatable by disease-associated matrix metalloproteinases. Hum Gene Ther. 1997;8:729–738.

    Article  CAS  Google Scholar 

  18. Peng KW, Vile R, Cosset FL, Russell S . Selective transduction of protease-rich tumors by matrix-metallo-proteinase-targeted retroviral vectors. Gene Therapy. 1999;6:1552–1557.

    Article  CAS  Google Scholar 

  19. Martin F, Kupsch J, Takeuchi Y, Russell S, Cosset FL, Collins M . Retroviral vector targeting to melanoma cells by single-chain antibody incorporation in envelope. Hum Gene Ther. 1998;9:737–746.

    Article  CAS  Google Scholar 

  20. Myers KA, Ryan MG, Stern PL, et al. Targeting immune effector molecules to human tumor cells through genetic delivery of 5T4-specific scFv fusion proteins. Cancer Gene Ther. 2002;9:884–896.

    Article  CAS  Google Scholar 

  21. Nguyen TH, Pages JC, Farge D, Briand P, Weber A . Amphotropic retroviral vectors displaying hepatocyte growth factor-envelope fusion proteins improve transduction efficiency of primary hepatocytes. Hum Gene Ther. 1998;9:2469–2479.

    CAS  PubMed  Google Scholar 

  22. Matsumoto K, Nakamura T . Hepatocyte growth factor-scatter factor (HGF-SF) and the c-Met receptor. In: Goldberg ID and Rosen EM, eds.; Hepatocyte Growth Factor-Scatter Factor (HGF-SF) and the c-Met Receptor. Basel/Switzerland: Birkhauser Verlag; 1993:225–249.

    Google Scholar 

  23. Grigioni WF, Fiorentino M, D'Errico A, Ponzetto A, Crepaldi T, Prat M, Comoglio PM . Overexpression of c-met proto-oncogene product and raised Ki67 index in hepatocellular carcinomas with respect to benign liver conditions. Hepatology. 1995;21:1543–1546.

    CAS  PubMed  Google Scholar 

  24. Ueki T, Nakata K, Mawatari F, et al. Retrovirus-mediated gene therapy for human hepatocellular carcinoma transplanted in athymic mice. Int J Mol Med. 1998;1: 671–675.

    CAS  PubMed  Google Scholar 

  25. Vassalli JD, Pepper MS . Tumour biology. Membrane proteases in focus. Nature 1994;370:14–15.

    Article  CAS  Google Scholar 

  26. Jeffers M, Rong S, Vande Woude GF . Enhanced tumorigenicity and invasion-metastasis by hepatocyte growth factor/scatter factor-met signalling in human cells concomitant with induction of the urokinase proteolysis network. Mol Cell Biol. 1996;16:1115–1125.

    Article  CAS  Google Scholar 

  27. Monvoisin A, Neaud V, De Ledinghen V, et al. Direct evidence that hepatocyte growth factor-induced invasion of hepatocellular carcinoma cells is mediated by urokinase. J Hepatol. 1999;30:511–518.

    Article  CAS  Google Scholar 

  28. Andreasen PA, Kjoller L, Christensen L, Duffy MJ . The urokinase-type plasminogen activator system in cancer metastasis: a review. Int J Cancer. 1997;72:1–22.

    Article  CAS  Google Scholar 

  29. Lavillette D, Russell SJ, Cosset FL . Retargeting gene delivery using surface-engineered retroviral vector particles. Curr Opin Biotechnol. 2001;12:461–466.

    Article  CAS  Google Scholar 

  30. Pages JC, Andreoletti M, Bennoun M, et al. Efficient retroviral-mediated gene transfer into primary culture of murine and human hepatocytes: expression of the LDL receptor. Hum Gene Ther. 1995;6:21–30.

    Article  CAS  Google Scholar 

  31. Ohashi K, Marion PL, Nakai H, et al. Sustained survival of human hepatocytes in mice: a model for in vivo infection with human hepatitis B and hepatitis delta viruses. Nat Med. 2000;6:327–331.

    Article  CAS  Google Scholar 

  32. Cosset FL, Morling FJ, Takeuchi Y, Weiss RA, Collins MK, Russell SJ . Retroviral retargeting by envelopes expressing an N-terminal binding domain. J Virol. 1995;69:6314–6322.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Towle MJ, Lee A, Maduakor EC, Schwartz CE, Bridges AJ, Littlefield BA . Inhibition of urokinase by 4-substituted benzo[b]thiophene-2-carboxamidines: an important new class of selective synthetic urokinase inhibitor. Cancer Res. 1993;53:2553–2559.

    CAS  PubMed  Google Scholar 

  34. Naldini L, Tamagnone L, Vigna E, et al. Extracellular proteolytic cleavage by urokinase is required for activation of hepatocyte growth factor/scatter factor. EMBO J. 1992;11:4825–4833.

    Article  CAS  Google Scholar 

  35. Andreoletti M, Loux N, Vons C, et al. Engraftment of autologous retrovirally transduced hepatocytes after intraportal transplantation into nonhuman primates: implication for ex vivo gene therapy. Hum Gene Ther. 2001;12:169–179.

    Article  CAS  Google Scholar 

  36. Prat M, Crepaldi T, Pennacchietti S, Bussolino F, Comoglio PM . Agonistic monoclonal antibodies against the Met receptor dissect the biological responses to HGF. J Cell Sci. 1998;111:237–247.

    CAS  PubMed  Google Scholar 

  37. Matsumoto K, Kataoka H, Date K, Nakamura T . Cooperative interaction between alpha- and beta-chains of hepatocyte growth factor on c-Met receptor confers ligand-induced receptor tyrosine phosphorylation and multiple biological responses. J Biol Chem. 1998;273:22913–22920.

    Article  CAS  Google Scholar 

  38. Paciucci R, Vila MR, Adell T, et al. Activation of the urokinase plasminogen activator/urokinase plasminogen activator receptor system and redistribution of E-cadherin are associated with hepatocyte growth factor-induced motility of pancreas tumor cells overexpressing Met. Am J Pathol. 1998;153:201–212.

    Article  CAS  Google Scholar 

  39. Ljubimova JY, Petrovic LM, Wilson SE, Geller SA, Demetriou AA . Expression of HGF, its receptor c-met, c-myc, and albumin in cirrhotic and neoplastic human liver tissue. J Histochem Cytochem. 1997;45:79–87.

    Article  CAS  Google Scholar 

  40. Morita Y, Hayashi Y, Wang Y, et al. Expression of urokinase-type plasminogen activator receptor in hepatocellular carcinoma. Hepatology. 1997;25:856–861.

    Article  CAS  Google Scholar 

  41. Nemunaitis J, Edelman J . Selectively replicating viral vectors. Cancer Gene Ther. 2002;9:987–1000.

    Article  CAS  Google Scholar 

  42. Solly SK, Trajcevski S, Frisen C, et al. Replicative retroviral vectors for cancer gene therapy. Cancer Gene Ther. 2003;10:30–39.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr François-Loïc Cosset for providing the expression vectors and the TELCeB6 and FlyTA7 cell lines, to Dr Bruce Littlefield for providing the uPA inhibitor B428. This work was supported in part by grants from Association pour la Recherche contre le Cancer, the European Community (QLK3 CT 1999), Fondation pour la Recherche Médicale, Université Paris XI, and by “Ministère de l'Education Nationale, de l'Enseignement supérieur et de la Recherche” (to THN) and by Association Française contre les Myopathies (to NL).

Author information

Author notes

  1. Tuan Huy Nguyen

    Present address: Department of Microbiology, Geneva School of Medecine, 1 Rue Michel Servet, CH-1211, Geneva, Switzerland

Authors and Affiliations

  1. INSERM EMI 00-20, Hôpital A. Béclère, 157 rue de la Porte de Trivaux, Clamart, 92141, France

    Tuan Huy Nguyen, Nathalie Loux, Ibrahim Dagher, Corinne Vons, Dominique Franco & Anne Weber

  2. Service de chirurgie générale, Hôpital A. Béclère, Clamart, 92141, France

    Ibrahim Dagher, Corinne Vons & Dominique Franco

  3. Department of Molecular Oncology, Genentech Inc., South San Francisco, 9408, California, USA

    Ken Carey & Ralph Schwall

  4. Institut Cochin, 22 rue Méchain, Paris, 75014, France

    Pascale Briand

  5. INSERM U. 347, Hôpital du Kremlin-Bicêtre, Kremlin-Bicêtre, France

    Michelle Hadchouel

  6. UMR144, Institut Curie, 26 rue d'Ulm, 75248, Paris Cedex 05, France

    Jacqueline Jouanneau

Authors
  1. Tuan Huy Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nathalie Loux
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ibrahim Dagher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Corinne Vons
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ken Carey
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pascale Briand
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Michelle Hadchouel
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Dominique Franco
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jacqueline Jouanneau
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ralph Schwall
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Anne Weber
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anne Weber.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nguyen, T., Loux, N., Dagher, I. et al. Improved gene transfer selectivity to hepatocarcinoma cells by retrovirus vector displaying single-chain variable fragment antibody against c-Met. Cancer Gene Ther 10, 840–849 (2003). https://doi.org/10.1038/sj.cgt.7700640

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.cgt.7700640

Keywords

This article is cited by

Search

Advanced search

Quick links