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Herpes simplex virus thymidine kinase/ganciclovir–induced cell death is enhanced by co-expression of caspase-3 in ovarian carcinoma cells

Cancer Gene Therapy volume 8, pages 308–319 (2001)Cite this article

Abstract

There is a need to enhance the efficacy of genetic prodrug activation therapy using herpes simplex virus thymidine kinase (tk) and ganciclovir (GCV) following disappointing results in early clinical trials. tk/GCV has been shown to lead to the activation of caspase-3, a potent executor of apoptosis. We demonstrate that co-expression of pro-caspase-3 with tk/GCV leads to enhanced cell death in ovarian carcinoma cells in vitro. Following transfection with recombinant adenoviral vectors encoding tk, GCV treatment leads to greater cell death in pro-caspase-3–expressing clones of SKOV3 and IGROV1 than control cells, as well as more rapid activation of caspase-3 and more rapid cleavage of PARP. Flow cytometry suggests that there is a greater degree of S-phase block in the pro-caspase-3–expressing clones than in control cells following treatment with tk/GCV. None of these effects is seen following transfection with a control adenovirus that does not encode tk. The increased cell death, early caspase-3 activation and PARP cleavage, and flow cytometric changes seen in pro-caspase-3–expressing cells can be partially inhibited by treatment with benzyloxycarbonyl–val–ala–asp fluoromethylketone, a synthetic caspase inhibitor. Our data suggest that co-expression of pro-caspase-3 may lead to a significant enhancement of the efficacy of tk/GCV therapy. Cancer Gene Therapy (2001) 8, 308–319

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References

  1. Huber B, Austin E, Richards C, et al . Metabolism of 5-fluorocytosine to 5-fluorouracil in human colorectal tumor cells transduced with the cytosine deaminase gene — significant antitumor effects when only a small percentage of tumor cells express cytosine deaminase Proc Natl Acad Sci USA 1994; 91:: 8302–8306

    Article  Google Scholar 

  2. McNeish I, Green N, Gilligan M, et al . Virus-directed enzyme prodrug therapy for ovarian and pancreatic cancer using retrovirally delivered E. coli nitroreductase and CB1954 Gene Ther 1998; 5:: 1061–1069

    Article  Google Scholar 

  3. Rainov N, Dobberstein K, Sena-Esteves M, et al . New prodrug activation gene therapy for cancer using cytochrome P450 4B1 and 2-aminoanthracene/4-ipomeanol Hum Gene Ther 1998; 9:: 1261–1273

    Article  Google Scholar 

  4. Cortés M, de Felipe P, Martin V, et al . Successful use of a plant gene in the treatment of cancer in vivo Gene Ther 1998; 5:: 1499–1507

    Article  Google Scholar 

  5. Kojima A, Hackett N, Ohwada A, et al . In vivo human carboxylesterase cDNA gene transfer to activate the prodrug CPT-11 for local treatment of solid tumors J Clin Invest 1998; 101:: 1789–1796

    Article  Google Scholar 

  6. Moolten F . Tumor chemosensitivity conferred by inserted herpes thymidine kinase genes: paradigm for a prospective cancer control strategy Cancer Res 1986; 46:: 5276–5281

    Google Scholar 

  7. Izquierdo M, Martin V, de Felipe P, et al . Human malignant brain tumor response to herpes simplex thymidine kinase (HSVtk)/ganciclovir gene therapy Gene Ther 1996; 3:: 491–495

    Google Scholar 

  8. Ram Z, Culver K, Oshiro E, et al . Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producer cells Nat Med 1997; 4:: 1354–1361

    Article  Google Scholar 

  9. Shand N, Weber F, Mariani L, et al . A phase 1–2 clinical trial of gene therapy for recurrent glioblastoma multiforme by tumor transduction with the herpes simplex thymidine kinase gene followed by ganciclovir Hum Gene Ther 1999; 10:: 2325–2335

    Article  Google Scholar 

  10. Klatzmann D, Valery C, Bensimon G, et al . A phase I/II study of herpes simplex virus type I thymidine kinase “suicide” gene therapy for recurrent glioblastoma Hum Gene Ther 1998; 9:: 2595–2604

    Google Scholar 

  11. Herman J, Adler H, Aguilar-Cordova E, et al . In situ gene therapy for adenocarcinoma of the prostate: a phase I clinical trial Hum Gene Ther 1999; 10:: 1239–1249

    Article  Google Scholar 

  12. Loubiere L, Tiraby M, Cazaux C, et al . The equine herpes virus 4 thymidine kinase leads to a superior ganciclovir cell killing than the human herpes virus 1 thymidine kinase Gene Ther 1999; 6:: 1638–1642

    Article  Google Scholar 

  13. Cannon J, Hamzeh F, Moore S, et al . Human herpes virus 8–encoded thymidine kinase and phosphotransferase homologues confer sensitivity of ganciclovir J Virol 1999; 73:: 4786–4793

    PubMed Central  Google Scholar 

  14. Kokoris M, Sabo P, Adman E, et al . Enhancement of tumor ablation by a selected HSV-1 thymidine kinase mutant Gene Ther 1999; 6:: 1415–1426

    Article  Google Scholar 

  15. Tong X, Engehausen D, Kaufman R, et al . Improvement of gene therapy for ovarian cancer by using acyclovir instead of ganciclovir in adenovirus-mediated thymidine kinase gene therapy Anticancer Res 1998; 18:: 713–718

    Google Scholar 

  16. Uckert W, Kammertons T, Haack K, et al . Double suicide gene (cytosine deaminase and herpes simplex virus thymidine kinase) but not single gene transfer allows reliable elimination of tumor cells in vivo Hum Gene Ther 1998; 9:: 855–865

    Article  Google Scholar 

  17. Chen S, Chen X, Wang Y, et al . Combination gene therapy for liver metastasis of colon carcinoma in vivo Proc Natl Acad Sci USA 1995; 92:: 2577–2581

    Article  Google Scholar 

  18. Benedetti S, Dimeco F, Pollo B, et al . Limited efficacy of the HSV-TK/GCV system for gene therapy of malignant gliomas and perspectives for the combined transduction of the interleukin-4 gene Hum Gene Ther 1997; 8:: 1345–1353

    Article  Google Scholar 

  19. Wei S-J, Chao Y, Shih Y-L, et al . Involvement of Fas (CD95/APO-1) and Fas ligand in apoptosis induced by ganciclovir treatment of tumor cells transduced with herpes simplex virus thymidine kinase Gene Ther 1999; 6:: 420–431

    Article  Google Scholar 

  20. Beltinger C, Fulda S, Kammertoens T, et al . 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  Google Scholar 

  21. Nicholson D . Caspase structure, proteolytic substrates, and function during apoptotic cell death Cell Death Differ 1999; 6:: 1028–1042

    Article  Google Scholar 

  22. Shinoura N, Murumatsa Y, Yoshida Y, et al . Adenovirus-mediated transfer of caspase-3 with Fas ligand induces drastic apoptosis in U-373MG glioma cells Exp Cell Res 2000; 256:: 423–433

    Article  Google Scholar 

  23. Yamabe K, Shimizu S, Ito T, et al . Cancer gene therapy using a pro-apoptotic gene, caspase-3 Gene Ther 1999; 6: 1952–1959

    Article  CAS  Google Scholar 

  24. Srinivasula S, Ahmad M, MacFarlane M, et al . Generation of constitutively active recombinant caspases-3 and -6 by rearrangement of their subunits J Biol Chem 1998; 273:: 10107–10111

    Article  Google Scholar 

  25. Tenev T, Marani M, McNeish I, et al . Caspase-3 overexpression sensitizes ovarian cancer cells to proteasome inhibitors Cell Death Differ 2000 In press

  26. Garcia-Calvo M, Peterson E, Leiting B, et al . Inhibition of human caspases by peptide-based and macromolecular inhibitors J Biol Chem 1998; 273:: 32608–32613

    Article  Google Scholar 

  27. Tenev T, Böhmer S-A, Kaufmann R, et al . Perinuclear localization of the protein-tyrosine phosphatase SHP-1 and inhibition of epidermal growth factor–stimulated STAT1/3 activation in A431 cells Eur J Cell Biol 2000; 79:: 261–271

    Article  Google Scholar 

  28. Vassaux G, Hurst H, Lemoine N . Insulation of a conditionally expressed transgene in an adenoviral vector Gene Ther 1999; 6:: 1192–1197

    Article  Google Scholar 

  29. Ring C, Blouin P, Martin L-A, et al . Use of transcriptional regulatory elements of the MUC1 and ERBB2 genes to drive tumor-selective expression of a prodrug activating enzyme Gene Ther 1997; 4:: 1045–1052

    Article  Google Scholar 

  30. Mosmann T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays J Immunol Methods 1983; 65:: 55–63

    Article  Google Scholar 

  31. Yu D, Wolf J, Scanlon M, et al . Enhanced c-erbB-2/neu expression in human ovarian cancer cells correlates with more severe malignancy that can be suppressed by E1A Cancer Res 1993; 53:: 891–898

    Google Scholar 

  32. Kovarik A, Peat N, Wilson D, et al . Analysis of the tissue-specific promoter of the MUC1 gene J Biol Chem 1993; 268:: 9917–9926

    Google Scholar 

  33. Lazebnik Y, Kaufmann S, Desnoyers S, et al . Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE Nature 1994; 371:: 346–347

    Article  Google Scholar 

  34. Nicholson DW, Ali A, Thornberry NA, et al . Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis Nature 1995; 376:: 37–43

    Article  Google Scholar 

  35. Decker T, Lohmannmatthes ML . A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity J Immunol Methods 1988; 115:: 61–69

    Article  Google Scholar 

  36. Jänicke R, Ng P, Sprengart M, et al . Caspase-3 is required for α-fodrin cleavage but dispensable for cleavage of other death substrates in apoptosis J Biol Chem 1998; 273:: 15540–15545

    Article  Google Scholar 

  37. Clarke R, Lund E, Johnson I, et al . Apoptosis can be detected in attached colonic adenocarcinoma HT29 cells using annexin V binding, but not by TUNEL assay or sub G0 DNA content Cytometry 2000; 39:: 141–150

    Article  Google Scholar 

  38. Melcher A, Todryk S, Hardwick S, et al . Tumor immunogenicity is determined by the mechanism of cell death via induction of heat shock protein expression Nat Med 1998; 4:: 581–587

    Article  Google Scholar 

  39. Laster S, Wood J, Gooding L . Tumor necrosis factor can induce both apoptotic and necrotic forms of cell lysis J Immunol 1988; 141:: 2629–2635

    Google Scholar 

  40. Saldeen J . Cytokines induce both necrosis and apoptosis via a common bcl-2 inhibitable pathway in rat insulin-producing cells Endocrinology 2000; 141:: 2003–2010

    Article  Google Scholar 

  41. Vercammen D, Brouckaert G, Denecker G, et al . Dual signalling of the Fas receptor: initiation of both apoptotic and necrotic cell death receptors J Exp Med 1998; 188: 919–930

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. ICRF Molecular Oncology Unit, Imperial College School of Medicine, Hammersmith Hospital, London, W12 0NN, UK

    I A McNeish, T Tenev, S Bell, M Marani, G Vassaux & N Lemoine

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  1. I A McNeish
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  2. T Tenev
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  3. S Bell
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  4. M Marani
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  5. G Vassaux
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  6. N Lemoine
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Correspondence to I A McNeish.

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McNeish, I., Tenev, T., Bell, S. et al. Herpes simplex virus thymidine kinase/ganciclovir–induced cell death is enhanced by co-expression of caspase-3 in ovarian carcinoma cells. Cancer Gene Ther 8, 308–319 (2001). https://doi.org/10.1038/sj.cgt.7700305

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