Tissue-specific expression of herpes simplex virus thymidine kinase gene delivered by adeno-associated virus inhibits the growth of human hepatocellular carcinoma in athymic mice.

About 70% of hepatocellular carcinomas are known to express alpha-fetoprotein, which is normally expressed in fetal but not in adult livers. To induce herpes simplex virus-thymidine kinase expression in these cancer cells, we constructed an adeno-associated viral vector containing the HSV-TK gene under the control of the alpha-fetoprotein enhancer and albumin promoter. We previously demonstrated in vitro that although this vector can transduce a variety of human cells, only transduced AFP and albumin-expressing hepatocellular carcinoma cell lines were sensitive to killing by ganciclovir (GCV). In the present study, we explored the effect of this vector on hepatocellular carcinoma cells in vivo. Subcutaneous tumors generated in nude mice by implanting hepatocellular carcinoma cells previously transduced with this vector shrank dramatically after treatment with GCV. Bystander effect was also observed on the tumors generated by mixing transduced and untransduced cells. To test whether the tumor cells can be transduced by the virus in vivo, we injected the recombinant adeno-associated virus into tumors generated by untransduced hepatocarcinoma cell line. Tumor growth were retarded after treatment with GCV. These experiments demonstrate the feasibility of in vivo transduction of tumor cell with rAAV.

[1]  J. Dong,et al.  Reactivation of silenced, virally transduced genes by inhibitors of histone deacetylase. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[2]  B. Dörken,et al.  Liver-associated toxicity of the HSV-tk/GCV approach and adenoviral vectors. , 1997, Cancer gene therapy.

[3]  Y. Kan,et al.  Selective killing of AFP-positive hepatocellular carcinoma cells by adeno-associated virus transfer of the herpes simplex virus thymidine kinase gene. , 1996, Human gene therapy.

[4]  V. Cherington,et al.  Tumor cells expressing the herpes simplex virus-thymidine kinase gene in the treatment of Walker 256 meningeal neoplasia in rats. , 1996, Journal of neurosurgery.

[5]  S. Kaneko,et al.  Adenovirus-mediated gene therapy of hepatocellular carcinoma using cancer-specific gene expression. , 1995, Cancer research.

[6]  Y. Kaneko,et al.  Gene therapy of hepatoma: bystander effects and non-apoptotic cell death induced by thymidine kinase and ganciclovir. , 1995, Cancer letters.

[7]  M. Zoppè,et al.  Generation of targeted retroviral vectors by using single-chain variable fragment: an approach to in vivo gene delivery. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Ramesh,et al.  The role of cytokines in mediating the bystander effect using HSV-TK xenogeneic cells. , 1995, Cancer letters.

[9]  G. Finocchiaro,et al.  The "bystander effect": association of U-87 cell death with ganciclovir-mediated apoptosis of nearby cells and lack of effect in athymic mice. , 1995, Human gene therapy.

[10]  Y. Kan,et al.  Tissue-specific targeting of retroviral vectors through ligand-receptor interactions. , 1994, Science.

[11]  T. Flotte,et al.  Adeno-associated virus vector gene expression occurs in nondividing cells in the absence of vector DNA integration. , 1994, American journal of respiratory cell and molecular biology.

[12]  I. Kawase,et al.  Gene therapy for carcinoembryonic antigen-producing human lung cancer cells by cell type-specific expression of herpes simplex virus thymidine kinase gene. , 1994, Cancer research.

[13]  J. Bertram,et al.  Re: Z. Ram et al., In situ retroviral-mediated gene transfer for the treatment of brain tumors in rats. Cancer Res., 53: 83-88, 1993. , 1994, Cancer research.

[14]  R. Warnick,et al.  In vitro evidence that metabolic cooperation is responsible for the bystander effect observed with HSV tk retroviral gene therapy. , 1993, Human gene therapy.

[15]  R. Vile,et al.  Use of tissue-specific expression of the herpes simplex virus thymidine kinase gene to inhibit growth of established murine melanomas following direct intratumoral injection of DNA. , 1993, Cancer research.

[16]  R. Martuza,et al.  Gene therapy of maliganant brain tumors: A rat glioma line bearing the herpes simplex virus type 1‐thymidine kinase gene and wild type retrovirus kills other tumor cells , 1992, Journal of neuroscience research.

[17]  Z. Ram,et al.  In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors. , 1992, Science.

[18]  H. Nakabayashi,et al.  A position-dependent silencer plays a major role in repressing alpha-fetoprotein expression in human hepatoma , 1991, Molecular and cellular biology.

[19]  C. Richards,et al.  Retroviral-mediated gene therapy for the treatment of hepatocellular carcinoma: an innovative approach for cancer therapy. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[20]  R. Martuza,et al.  Selective killing of glioma cells in culture and in vivo by retrovirus transfer of the herpes simplex virus thymidine kinase gene. , 1991, The New biologist.

[21]  F. Moolten,et al.  Curability of tumors bearing herpes thymidine kinase genes transferred by retroviral vectors. , 1990, Journal of the National Cancer Institute.

[22]  Samulski,et al.  Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression , 1989, Journal of virology.

[23]  D. Richman,et al.  Thymidine kinase obliteration: creation of transgenic mice with controlled immune deficiency. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[24]  F. Moolten.  Tumor chemosensitivity conferred by inserted herpes thymidine kinase genes: paradigm for a prospective cancer control strategy. , 1986, Cancer research.

[25]  E W Geddes,et al.  Establishment of a continuously growing cell line from primary carcinoma of the liver. , 1976, South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde.