HSV-tk/IL-2 Gene Therapy for Glioblastoma Multiforme

  • Luisa Barzon
  • Monia Pacenti
  • Elisa Franchin
  • Federico Colombo
  • Giorgio PalùEmail author
Part of the Methods in Molecular Biology™ book series (MIMB, volume 542)


We performed a clinical trial of gene therapy in patients with recurrent glioblastoma multiforme based on retroviral vector-mediated combined delivery of interleukin-2 and thymidine kinase of herpes simplex virus (HSV)-1. Treatment consisted of intratumor injection of retroviral vector-producing cells, followed by intravenous ganciclovir administration. The primary endpoints of the study were the evaluation of the safety of the treatment and demonstration of transduction of tumor cells. Here, we report details on the clinical protocol and the methods used.

Key Words

Clinical trial gene therapy glioblastoma multiforme interleukin-2 retroviral vector suicide gene thymidine kinase 



This work was supported by grant no. RSF 271/07 from Regione Veneto to Prof. Giorgio Palù.


  1. 1.
    Barzon, L., Zanusso, M., Colombo, F., Palù, G. (2006) Clinical trials of gene therapy, virotherapy, and immunotherapy for malignant gliomas. Cancer Gene Ther. 13, 539–554.PubMedCrossRefGoogle Scholar
  2. 2.
    Rainov, N.G. (2000) A phase III clinical evaluation of herpes simplex virus type 1 thymidine kinase and ganciclovir gene therapy as an adjuvant to surgical resection and radiation in adults with previously untreated glioblastoma multiforme. Hum. Gene Ther. 11, 2389–2401.PubMedCrossRefGoogle Scholar
  3. 3.
    Rainov, N.G., Kramm, C.M., Banning, U., Riemann, D., Holzhausen, H.J., Heidecke, V., Burger, K.J., Burkert, W., Körholz, D. (2000) Immune response induced by retrovirus-mediated HSV-tk/GCV pharmacogene therapy in patients with glioblastoma multiforme. Gene Ther. 7, 1853–1858.PubMedCrossRefGoogle Scholar
  4. 4.
    Pizzato, M., Franchin, E., Calvi, P., Boschetto, R., Colombo, M., Ferrini, S., Palù, G. (1998) Production and characterization of a bicistronic Moloney-based retroviral vector expressing human interleukin 2 and herpes simplex virus thymidine kinase for gene therapy of cancer. Gene Ther. 5, 1003–1007.PubMedCrossRefGoogle Scholar
  5. 5.
    Barzon, L., Bonaguro, R., Castagliuolo, I., Chilosi, M., Gnatta, E., Parolin, C., Boscaro, M., Palù, G. (2002) Transcriptionally targeted retroviral vector for combined suicide and immunomodulating gene therapy of thyroid cancer. J. Clin. Endocrinol. Metab. 87, 5304–5311.PubMedCrossRefGoogle Scholar
  6. 6.
    Barzon, L., Bonaguro, R., Castagliuolo, I., Chilosi, M., Franchin, E., Del Vecchio, C., Giaretta, I., Boscaro, M., Palù, G. (2003) Gene therapy of thyroid cancer via retrovirally-driven combined expression of human interleukin-2 and herpes simplex virus thymidine kinase. Eur J Endocrinol. 148, 73–80.PubMedCrossRefGoogle Scholar
  7. 7.
    Stefani, A.L., Barzon, L., Castagliuolo, I., Guido, M., Pacenti, M., Parolin, C., Farinati, F., Palù, G. (2005) Systemic efficacy of combined suicide/cytokine gene therapy in a murine model of hepatocellular carcinoma. J Hepatol. 42, 728–35.PubMedCrossRefGoogle Scholar
  8. 8.
    Palu, G., Cavaggioni, A., Calvi, P., Franchin, E., Pizzato, M., Boschetto, R., Parolin, C., Chilosi, M., Ferrini, S., Zanusso, A., Colombo, F. (1999) Gene therapy of glioblastoma multiforme via combined expression of suicide and cytokine genes: a pilot study in humans. Gene Ther. 6, 330–337.PubMedCrossRefGoogle Scholar
  9. 9.
    Colombo, F., Zanusso, M., Casentini, L., Cavaggioni, A., Franchin, E., Calvi, P., Palù, G. (1997) Gene stereotactic neurosurgery for recurrent malignant gliomas. Stereotact. Funct. Neurosurg. 68, 245–251.PubMedCrossRefGoogle Scholar
  10. 10.
    Colombo, F., Barzon, L., Franchin, E., Pacenti, M., Pinna, V., Danieli, D., Zanusso, M., Palù, G. (2005) Combined HSV-TK/IL-2 gene therapy in patients with recurrent glioblastoma multiforme: biological and clinical results. Cancer Gene Ther. 12, 835–848.PubMedCrossRefGoogle Scholar
  11. 11.
    Barzon, L., Pacenti, M., Taccaliti, A., Franchin, E., Bruglia, M., Boscaro, M., Palù, G. (2005) A pilot study of combined suicide/cytokine gene therapy in two patients with end-stage anaplastic thyroid carcinoma. J Clin Endocrinol Metab. 90:2831–2834.PubMedCrossRefGoogle Scholar
  12. 12.
    Oldfield, E.H., Ram, Z., Culver, K.W., Blaese, R.M., DeVroom, H.L., Anderson, W.F. (1993) Gene therapy for the treatment of brain tumors using intra-tumoral transduction with the thymidine kinase gene and intravenous ganciclovir. Hum Gene Ther. 4, 39–69.PubMedCrossRefGoogle Scholar
  13. 13.
    Palù, G., Pizzato, M., Bonaguro, R., Colombo, F. (2000) Gene therapy of glioblastoma multiforme with a bicistronic retroviral vector expressing human IL-2 and HSV-TK, in Gene Therapy of Cancer: Methods and Protocols (Walther, W., Stein, U. ed.), Humana Press, Totowa, NJ, pp. 511–522.Google Scholar
  14. 14.
    Barzon, L., Gnatta, E., Castagliuolo, I., Trevisan, M., Moretti, F., Pontecorvi, A., Boscaro, M., Palù, G. (2005) Modulation of retrovirally driven therapeutic genes by mutant TP53 in anaplastic thyroid carcinoma. Cancer Gene Ther. 12, 381–388.PubMedCrossRefGoogle Scholar
  15. 15.
    Li, Z., Düllmann, J., Schiedlmeier, B., Schmidt, M., von Kalle, C., Meyer, J., Forster, M., Stocking, C., Wahlers, A., Frank, O., Ostertag, W., Kühlcke, K., Eckert, H.G., Fehse, B., Baum, C. (2002) Murine leukemia induced by retroviral gene marking. Science 296, 497.PubMedCrossRefGoogle Scholar
  16. 16.
    Modlich, U., Kustikova, O.S., Schmidt, M., Rudolph, C., Meyer, J., Li, Z., Kamino, K., von Neuhoff, N., Schlegelberger, B., Kuehlcke, K., Bunting, K.D., Schmidt, S., Deichmann, A., von Kalle, C., Fehse, B., Baum, C. (2005) Leukemias following retroviral transfer of multidrug resistance 1 (MDR1) are driven by combinatorial insertional mutagenesis. Blood 105, 4235–4246.PubMedCrossRefGoogle Scholar
  17. 17.
    Hacein-Bey-Abina, S., Von Kalle, C., Schmidt, M., McCormack, M.P., Wulffraat, N., Leboulch, P., Lim, A., Osborne, CS., Pawliuk, R., Morillon, E., Sorensen, R., Forster, A., Fraser, P., Cohen, JI., de Saint Basile, G., Alexander, I., Wintergerst, U., Frebourg T., Aurias A., Stoppa-Lyonnet D., Romana S., Radford-Weiss I., Gross F., Valensi F., Delabesse E., Macintyre E., Sigaux F., Soulier J., Leiva LE., Wissler M., Prinz C., Rabbitts TH., Le Deist F., Fischer A., Cavazzana-Calvo M. (2003) LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 302, 415–419.PubMedCrossRefGoogle Scholar
  18. 18.
    Couzin, J., Kaiser, J. (2005) As Gelsinger Case Ends, Gene Therapy Suffers Another Blow. Science 307, 1028.PubMedCrossRefGoogle Scholar
  19. 19.
    Trasher, A., Gaspar, B. Severe Adverse Event in Clinical Trial of Gene Therapy for X-SCID. ( Dec 18th, 2007.
  20. 20.
    U.S. Department of Health and Human Services Food and Drug Administration Center for Biologics Evaluation and Research. Guidance for Industry: Gene Therapy Clinical Trials – Observing Subjects for Delayed Adverse Events. November 2006. (

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Luisa Barzon
    • 1
  • Monia Pacenti
    • 1
  • Elisa Franchin
    • 1
  • Federico Colombo
    • 2
  • Giorgio Palù
    • 1
    Email author
  1. 1.Department of Histology, Microbiology and Medical BiotechnologiesUniversity of PadovaPadovaItaly
  2. 2.Division of Neurosurgery, Robotic Radiosurgery UnitSan Bortolo HospitalVicenzaItaly

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