Abstract
Implantation of retrovirus-producing cells within a tumor has been demonstrated to eliminate malignant brain tumors effectively in animal models. In our previous study, the implantation of high-titer retrovirus-producing fibroblasts into tumors resulted in highly efficient transduction in vivo. The transduced glioma cells migrated far from the implantation site, potentiating the induction of the remarkable bystander effect. It is also possible, however, that the implantation of murine fibroblast-derived virus-producing cells may induce an immune response in patients. In this study, we prepared retroviruses carrying the herpes simplex virus thymidine kinase (HTK) gene with titers of 1.4–2.5 × 1011 colony-forming units (c.f.u.)/ml, and stereotactically inoculated only 3 μl of the HTK-bearing retroviruses into the brain tumors of mice. Following repetitive ganciclovir (GCV) intraperitoneal injection, effective killing of glioma cells in the mouse brain was observed. The transduction efficiency was nearly as high as that observed for the implantation of high-titer retrovirus-producing fibroblasts. Eighty percent of brain tumor-bearing mice were completely cured by our treatment protocol using concentrated HTK-harboring retroviruses. Our results suggest that repeated inoculations of high-titer retroviruses carrying the HTK gene followed by GCV treatment may be a promising strategy for the clinical treatment of malignant gliomas.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Russell D, Rubinstein L . Pathology of Tumors of the Nervous Sysem Edward Arnold: London 1989 pp 83–247
Moolten FL . Tumor chemosensitivity conferred by inserted herpes thymidine kinase genes: paradigm for aprospective cancer control strategy Cancer Res 1986 46: 5276–5281
Moolten FL, Wells JM . Curability of tumors bearing herpes thymidine kinase genes transfected by retroviral vectors J Natl Cancer Inst 1990 82: 297–300
Kramm CM et al. Gene therapy for brain tumors Brain Pathol 1995 5: 345–381
Miller DG, Adam MA, Miller AD . Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection Mol Cell Biol 1990 10: 4239–4242
Short MP et al. Gene delivery to glioma cells in rat brain by grafting of retrovirus packaging cell line J Neurosci Res 1990 27: 427–439
Tamiya T et al. Transgene inheritance and retroviral infection contribute to the efficiency of gene expression in solid tumors inoculated with retroviral vector producer cells Gene Therapy 1995 2: 531–538
Yoshimatsu T, Tamura M, Kuriyama S, Ikenaka K . Improvement of retroviral packaging cell lines by introducing the polyoma virus early region Hum Gene Ther 1998 9: 161–172
Tamura M et al. Transduction of glioma cells using a high titer retroviral vector system and their subsequent migration in brain tumors Gene Therapy 1998 5: 1698–1704
Yamada M et al. Migration of genetically labeled glioma cells after implantation into murine brain J Neurosci Res 1994 38: 415–423
Tamura M et al. Targeted killing of migrating glioma cells by injection of HTK-modified glioma cells Hum Gene Ther 1997 8: 381–391
Kuriyama S et al. Complete cure of established murine hepatocellular carcinoma is achievable by repeated injections of retroviruses carrying the herpes simplex virus thymidine kinase gene Gene Therapy 1999 6: 525–533
Kumanishi T, Ikuta F, Yamamoto T . Brain tumors induced by Rous sarcoma virus, Schmidt-Ruppin strain: III. Morphology of brain tumors inducced in adult mice J Natl Cancer Inst 1973 50: 95–109
Oldfield EH et al. Gene therapy for the treatment of brain tumors using intra-tumoral transduction with the thymidine kinase gene and intravenous ganciclovir Hum Gene Ther 1993 4: 39–69
Raffel C et al. Gene therapy for the treatment of recurrent pediatric malignant astrocytomas with in vivo tumor transduction with the herpes simplex thymidine kinase gene/ganciclovir system Hum Gene Ther 1994 5: 863–890
Ram Z et al. Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells Nature Med 1997 3: 1354–1361
Culver KW et al. In vivo gene transfer with retroviral vector producer cells for treatment of experimental brain tumors Science 1992 256: 1550–1552
Bi WL, Parysek LM, Warnick R, Stambrook PJ . In vivo evidence that metabolic cooperation is responsible for the bystander effect observed with HSVtk retroviral gene therapy Hum Gene Ther 1993 4: 725–731
Barba D, Hardin J, Sadelain M, Gage FH . Development of antitumor immunity following thymidine kinase-mediated killing of experimental brain tumors Proc Natl Acad Sci USA 1994 91: 4348–4352
Ram Z et al. In situ retroviral-mediated gene transfer for the treatment of brain tumors in rats Cancer Res 1993 53: 83–88
Sambrook J, Fritsch EF, Maniatis T . Molecular Cloning. A Laboratory Manual Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY 1989
Miyao Y et al. Selective expression of foreign genes in glioma cells: use of the mouse myelin basic protein gene promoter to direct toxic gene expression J Neurosci Res 1993 36: 472–479
Kuriyama S et al. Gene therapy for the treatment of hepatoma by retroviral-mediated gene transfer of the herpes simplex virus thymidine kinase Int Hepatol Commun 1993 1: 253–259
Bender MA, Palmer TD, Gelinas RE, Miller AD . Evidence that the packaging signal of Moloney murine leukemia virus extends into the gag region J Virol 1987 61: 1639–1646
Ikenaka K et al. Detection of brain-specific gene expression in brain cells in primary culture: a novel promotor assay based on the use of a retrovirus vector New Biol 1992 53: 53–60
Mann R, Mulligan RC, Baltimore D . Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus Cell 1983 33: 153–159
Cepko C . Preparation of a specific retrovirus producer cell line. In: Ausubel FM et al (eds) Current Protocols in Molecular Biology, Supplement 17 John Wiley: New York 1992 9.12.1–9.12.5
Bowles NE et al. A simple and efficient method of recombinant retrovirus for increased hepatocyte transduction in vivo Hum Gene Ther 1996 7: 1735–1742
Acknowledgements
This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tamura, K., Tamura, M., Ikenaka, K. et al. Eradication of murine brain tumors by direct inoculation of concentrated high titer-recombinant retrovirus harboring the herpes simplex virus thymidine kinase gene. Gene Ther 8, 215–222 (2001). https://doi.org/10.1038/sj.gt.3301371
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3301371