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Adenovirus E1a protein enhances the cytotoxic effects of the herpes thymidine kinase-ganciclovir system

Cancer Gene Therapy volume 10, pages 152–160 (2003)Cite this article

Abstract

Cancer gene therapy based on the use of suicide genes, such as the thymidine kinase gene, is not producing satisfactory results. Several approaches have been delineated to enhance the therapeutic responses, including augmentation of the bystander effect, the combination of the herpes simplex virus thymidine kinase-ganciclovir (HSVTK-GCV) system into replication competent adenoviruses and others. Moreover, because usually less than 20% of human malignant cells are in S-phase, the HSVTK-GCV system is not as efficient as expected. To increase the cytotoxic effects of the HSVTK-GCV system, we hypothesized that concomitant expression of E1a protein, which drives cells to proliferation and S-phase, could increase the effects of the HSVTK-GCV system. Several retroviruses were constructed carrying bicistronic sequences of TK and E1a 12S genes under the control of the CMV promoter. The constructions were tested in murine (NIH-3T3, MSC11A5) and human cells (IMR90, HeLa, MDA-MB435). A clear increase of the HSVTK-GCV system killing effect in nonconfluent cells was observed in the cells studied, especially in NIH-3T3, MSC11A5, IMR90, and MDA-MB435 expressing cells. In confluence, the NIH3T3 and IMR90 E1a-TK-expressing cells were also very sensitive and most malignant E1a-TK-expressing cells showed an irreversible G2-M cell cycle arrest. Moreover, the concomitant expression of adenovirus E1a and the HSVTK-GCV system increased the sensitivity to anticancer agents such as cisplatin. These results show that adenovirus E1a protein expression clearly enhances the cytotoxic effects of the HSVTK-GCV system and the response to treatment with cisplatin.

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References

  1. Springer CJ, Niculescu-Duvaz I . Prodrug-activating systems in suicide gene therapy. J Clin Invest. 2000;105:1161–1167.

    Article  CAS  Google Scholar 

  2. Wildner O, Morris JC . Therapy of peritoneal carcinomatosis from colon cancer with oncolytic adenoviruses. J Gene Med. 2000;2:353–360.

    Article  CAS  Google Scholar 

  3. Wildner O, Morris JC . The role of the E1B 55 kDa gene product in oncolytic adenoviral vectors expressing herpes simplex virus-TK: assessment of antitumor efficacy and toxicity. Cancer Res. 2000;60:4167–4174.

    CAS  PubMed  Google Scholar 

  4. Alemany R, Balague C, Curiel D . Replicative adenoviruses for cancer therapy. Nat Biotechnol. 2000;7:723–727.

    Article  Google Scholar 

  5. Heise C, Kirn DH . Replication-selective adenoviruses as oncolytic agents. J Clin Invest. 2000;7:847–851.

    Article  Google Scholar 

  6. Bischoff JR, Kirn DH, Williams A, et al. An adenovirus mutant that replicates selectively in p53 deficient human tumor cells. Science. 1996;274:373–376.

    Article  CAS  Google Scholar 

  7. lsley DD, Lee SH, Miller WH, Kuchta RD . Acyclic guanosine analogs inhibit DNA polymerases alpha, delta, and epsilon with very different potencies and have unique mechanisms of action. Biochemistry. 1995;34:2504–2510.

    Article  Google Scholar 

  8. Miller WH, Miller RL . Phosphorylation of acyclovir (acycloguanosine) monophosphate by GMP kinase. J Biol Chem. 1980;255:7204–7207.

    CAS  PubMed  Google Scholar 

  9. Frank KB, Chiou JF, Cheng YC . Interaction of herpes simplex virus-induced DNA polymerase with 9-(1,3-dihydroxy-2-propoxymethyl)guanine triphosphate. J Biol Chem. 1984;259:1566–1569.

    CAS  PubMed  Google Scholar 

  10. Halloran PJ, Fenton RG . Irreversible G2-M arrest and cytoskeletal reorganization induced by cytotoxic nucleoside analogues. Cancer Res. 1998;58:3855–3865.

    CAS  PubMed  Google Scholar 

  11. Sarisky RT, Quail MR, Clark PE, et al. Characterization of herpes simplex viruses selected in culture for resistance to penciclovir or acyclovir. J Virol. 2001;75:1761–1769.

    Article  CAS  Google Scholar 

  12. Tomicic MT, Thust R, Sobol RW, Kaina B . DNA polymerase beta mediates protection of mammalian cells against ganciclovir-induced cytotoxicity and DNA breakage. Cancer Res. 2001;61:7399–7403.

    CAS  PubMed  Google Scholar 

  13. Touraine RL, Vahanian N, Ramsey WJ, Blaese RM . Enhancement of the herpes simplex virus thymidine kinase/ganciclovir bystander effect and its antitumor efficacy in vivo by pharmacologic manipulation of gap junctions. Hum Gene Ther. 1998;9:2385–2391.

    Article  CAS  Google Scholar 

  14. Mesnil M, Yamasaki H . Bystander effect in herpes simplex virus-thymidine kinase/ganciclovir cancer gene therapy: role of gap-junctional intercellular communication. Cancer Res. 2000;60:3989–3999.

    CAS  PubMed  Google Scholar 

  15. Carrio M, Mazo A, Lopez-Iglesias C, Estivill X, Fillat C . Retrovirus-mediated transfer of the herpes simplex virus thymidine kinase and connexin26 genes in pancreatic cells results in variable efficiency on the bystander killing: implications for gene therapy. Int J Cancer. 2001;94:81–88.

    Article  CAS  Google Scholar 

  16. Aghi M, Kramm CM, Chou TC, Breakefield XO, Chiocca EA . Synergistic anticancer effects of ganciclovir/thymidine kinase and 5-fluorocytosine/cytosine deaminase gene therapies. J Natl Cancer Inst. 1998;90:370–380.

    Article  CAS  Google Scholar 

  17. Wildner O, Blaese RM, Morris JC . Synergy between the herpes simplex virus TK/ganciclovir prodrug suicide system and the topoisomerase I inhibitor topotecan. Hum Gene Ther. 1999;10:2679–2687.

    Article  CAS  Google Scholar 

  18. Lee YJ, Galoforo SS, Battle P, Lee H, Corry PM, Jessup JM . Replicating adenoviral vector-mediated transfer of a heat-inducible double suicide gene for gene therapy. Cancer Gene Ther. 2001;6:397–404.

    Article  Google Scholar 

  19. Barbeau D, Charbonneau R, Whalen SG, Bayley ST, Branton PE . Functional interactions within adenovirus E1a protein complexes. Oncogene. 1994;9:359–373.

    CAS  PubMed  Google Scholar 

  20. Frisch MS, Mymryk JS . Adenovirus-5 E!A: paradox and paradigm. Nat Rev Mol Cell Biol. 2002;3:441–452.

    Article  CAS  Google Scholar 

  21. Frisch SM . Tumor suppression activity of adenovirus E1a protein: anoikis and the epithelial phenotype. Adv Cancer Res. 2001;80:39–49.

    Article  CAS  Google Scholar 

  22. Sánchez-Prieto R, Quintanilla M, Cano A, et al. Carcinoma cell lines become sensitive to DNA-damaging agents by the expression of the adenovirus E1a gene. Oncogene. 1996;13:1083–1092.

    PubMed  Google Scholar 

  23. Xing X, Yujiao Chang J, Hung M . Preclinical and clinical study of HER-2/neu-targeting cancer gene therapy. Adv Drug Deliv Rev. 1998;30:219–227.

    Article  CAS  Google Scholar 

  24. Martin-Duque, et al. Adenovirus lacking 19-kDa and 55-kDa E1B genes exerts a marked cytotoxic effect in human malignant cells. Cancer Gene Ther. 1999;6:554–563.

    Article  Google Scholar 

  25. Wei SJ, et al. S- and G2-phase cell cycle arrests and apoptosis induced by ganciclovir in murine melanoma cells transduced with herpes simplex virus thymidine kinase. Exp Cell Res. 1998;241:66–75.

    Article  CAS  Google Scholar 

  26. Halloran PJ, Fenton RG . Irreversible G2-M arrest and cytoskeletal reorganization induced by cytotoxic nucleoside analogues. Cancer Res. 1998;58:3855–3865.

    CAS  PubMed  Google Scholar 

  27. Beltinger C, Fulda S, Kammertoens T, Meyer E, Uckert W, Debatin KM . Herpes simplex virus thymidine kinase/ganciclovir-induced apoptosis involves ligand-independent death receptor aggregation and activation of caspases. Proc Natl Acad Sci USA. 1999;96:8699–8704.

    Article  CAS  Google Scholar 

  28. Yageta M, Tsunoda H, Yamanaka T, et al. The adenovirus E1a domains required for induction of DNA rereplication in G2/M arrested cells coincide with those required for apoptosis. Oncogene. 1999;18:4767–4776.

    Article  CAS  Google Scholar 

  29. Wildner O, Blaese RM, Morris JC . Therapy of colon cancer with oncolytic adenovirus is enhanced by the addition of herpes simplex virus-thymidine kinase. Cancer Res. 1999;59:410–413.

    CAS  PubMed  Google Scholar 

  30. Lambright ES, Amin K, Wiewrodt R, et al. Inclusion of the herpes simplex thymidine kinase gene in a replicating adenovirus does not augment antitumor efficacy. Gene Ther. 2001;12:946–953.

    Article  Google Scholar 

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Acknowledgements

We thank Marta Izquierdo for providing the IRES-TK plasmid and Ramón Alemany and Silvio Gutkind for advice, support, and critical reading of the manuscript. We thank Martin Hadley-Adams for preparing the manuscript and Ruth Valero for technical work. This work was supported by grants from: the “Fondo Español de Investigaciones Sanitarias,” FIS 99/0504; the Comunidad de Madrid, CAM 98/08.1/2, Aventis; and the Areces Foundation. V.F.S. is supported by a postdoctoral fellowship from “Consejeria de Educación (CAM)” with participation of “Fondo Social Europeo.” C.P.C. is supported by a predoctoral fellowship from the “Fondo Español de Investigaciones Sanitarias.”

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Authors and Affiliations

  1. Department of Pathology, Clínica Puerta de Hierro, Madrid, Spain

    Carlos Parada, Javier Hernández Losa, Juan Guinea, Víctor Sánchez-Arévalo, Víctor Fernández Soria, Ricardo Sánchez-Prieto & Santiago Ramón y Cajal

  2. Department of Immunology, Clínica Puerta de Hierro, Madrid, Spain

    Luis Alvarez-Vallina

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  1. Carlos Parada
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Correspondence to Santiago Ramón y Cajal.

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Parada, C., Hernández Losa, J., Guinea, J. et al. Adenovirus E1a protein enhances the cytotoxic effects of the herpes thymidine kinase-ganciclovir system. Cancer Gene Ther 10, 152–160 (2003). https://doi.org/10.1038/sj.cgt.7700550

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