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Quantification and characterization of the bystander effect in prostate cancer cells following adenovirus-mediated FasL expression

Cancer Gene Therapy volume 10, pages 330–339 (2003)Cite this article

Abstract

Inducing Fas-mediated apoptosis in prostate cancer (PCa) is a promising new therapeutic approach with the potential to overcome delivery issues currently problematic in cancer gene therapy. We have previously demonstrated that a Fas Ligand (FasL) expressing adenovirus (AdGFPFasLTET) was able to induce Fas-mediated apoptosis in a panel of PCa cell lines regardless of their Fas-sensitivity as determined by the agonistic Fas antibody CH-11. We now report that AdGFPFasLTET-infected cells produce apoptotic bodies and cellular debris that continues to elicit FasL-mediated bystander killing in uninfected neighboring cells. Using light microscopy, we demonstrate that AdGFPFasLTET-infected cells release apoptotic bodies and cellular debris into the local environment and that this material will induce bystander killing in Jurkat, PPC-1, and PC-3 target cells, but not in DU145 and K-562 cells. The bystander killing mechanism is mediated through Fas/FasL interaction because it is significantly inhibited if target cells are pretreated with the pan spectrum caspase inhibitor Z-VAD-FMK or the Fas neutralizing antibody ZB-4. Coincubation of PPC-1 target cells with apoptotic bodies and cellular debris (effector material) induce nearly complete target cell killing at a ratio of 1:1 target to effector. Collectively, these data indicate that AdGFPFasLTET-infected PCa cells release apoptotic and cellular debris capable of inducing bystander killing in PCa and supports the development of FasL as a gene therapy agent.

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References

  1. Heise C, Ganly I, Kim YT, et al. Efficacy of a replication-selective adenovirus against ovarian carcinomatosis is dependent on tumor burden, viral replication and p53 status. Gene Ther. 2000;7:1925–1926.

    Article  CAS  Google Scholar 

  2. Haviv YS, Curiel DT . Conditional gene targeting for cancer gene therapy. Adv Drug Deliv Rev. 2001;53:135–154.

    Article  CAS  Google Scholar 

  3. Galanis E, Vile R, Russell SJ . Delivery systems intended for in vivo gene therapy of cancer: targeting and replication competent viral vectors. Crit Rev Oncol Hematol. 2001;38:177–192.

    Article  CAS  Google Scholar 

  4. Suda T, Takahashi T, Golstein P, et al. Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell. 1993;75:1169–1178.

    Article  CAS  Google Scholar 

  5. Yonehara S, Ishii A, Yonehara M . A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor. J Exp Med. 1989;169:1747–1756.

    Article  CAS  Google Scholar 

  6. Trauth BC, Klas C, Peters AM, et al. Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science. 1989;245:301–305.

    Article  CAS  Google Scholar 

  7. Hyer ML, Voelkel-Johnson C, Rubinchik S, et al. Intracellular fas ligand expression causes fas-mediated apoptosis in human prostate cancer cells resistant to monoclonal antibody-induced apoptosis. Mol Ther. 2000;2:348–358.

    Article  CAS  Google Scholar 

  8. Hyer ML, Sudarshan S, Kim Y, et al. Down-regulation of c-FLIP sensitizes DU145 prostate cancer cells to Fas-mediated apoptosis. Cancer Biol Ther. 2002;1:405–410.

    Article  Google Scholar 

  9. Rubinchik S, Ding R, Qiu A J, et al. Adenoviral vector which delivers FasL-GFP fusion protein regulated by the tet-inducible expression system. Gene Ther. 2000;7:875–885.

    Article  CAS  Google Scholar 

  10. Schneider P, Holler N, Bodmer JL, et al. Conversion of membrane-bound Fas(CD95) ligand to its soluble form is associated with downregulation of its proapoptotic activity and loss of liver toxicity. J Exp Med. 1998;187:1205–1213.

    Article  CAS  Google Scholar 

  11. Douglas JT, Rogers BE, Rosenfeld ME, et al. Targeted gene delivery by tropism-modified adenoviral vectors. Nat Biotechnol. 1996;14:1574–1578.

    Article  CAS  Google Scholar 

  12. Hedlund TE, Meech SJ, Srikanth S, et al. Adenovirus-mediated expression of Fas ligand induces apoptosis of human prostate cancer cells. Cell Death Differ. 1999;6:175–182.

    Article  CAS  Google Scholar 

  13. Jodo S, Xiao S, Hohlbaum A, et al. Apoptosis-inducing membrane vesicles. A novel agent with unique properties. J Biol Chem. 2001;276:39938–39944.

    Article  CAS  Google Scholar 

  14. Beltinger C, Fulda S, Kammertoens T, et al. Herpes simplex virus thymidine kinase/gancyclovir-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 

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

    Article  CAS  Google Scholar 

  16. Hall SJ, Canfield SE, Yan Y, et al. A novel bystander effect involving tumor cell-derived Fas and FasL interactions following Ad.HSV-tk and Ad.mIL-12 gene therapies in experimental prostate cancer. Gene Ther. 2002;9:511–517.

    Article  CAS  Google Scholar 

  17. Fukazawa T, Fujiwara T, Morimoto Y, et al. Differential involvement of the CD95 (Fas/APO-1) receptor/ligand system on apoptosis induced by the wild-type p53 gene transfer in human cancer cells. Oncogene. 1999;18:2189–2199.

    Article  CAS  Google Scholar 

  18. Eaton JD, Perry MJA, Todryk SM, et al. Genetic prodrug activation therapy (GPAT) in two rat prostate models generates an immune bystander effect and can be monitored by magnetic resonance techniques. Gene Ther. 2001;8:557–567.

    Article  CAS  Google Scholar 

  19. Rubinchik S, Wang D, Yu H, et al. A complex adenovirus vector that delivers FasL-GFP with combined prostate-specific and tetracycline-regulated expression. Mol Ther. 2001;4:416–426.

    Article  CAS  Google Scholar 

  20. Lowe SL, Rubinchik S, Honda Y, et al. Prostate-specific expression of Bax delivered by an adenoviral vector induces apoptosis in LNCaP prostate cancer cells. Gene Ther. 2001;8:1363–1371.

    Article  CAS  Google Scholar 

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Acknowledgements

Supported by NIH CA 69596,CA 88163 and DOD Grant N6311601MD10004. We thank Rick Peppler and the MUSC flow cytometry facility for acquiring the flow cytometry data. We especially thank Joanne Douglas for kindly providing the adenovirus neutralizing antibody 1D6.14.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, 29425, South Carolina, USA

    Marc L Hyer, David A Schwartz, Jian-yun Dong & James S Norris

  2. Department of Urology, Medical University of South Carolina, Charleston, 29425, South Carolina, USA

    Sunil Sudarshan

  3. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, 29425, South Carolina, USA

    Yusuf Hannun

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  1. Marc L Hyer
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  2. Sunil Sudarshan
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Correspondence to James S Norris.

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Hyer, M., Sudarshan, S., Schwartz, D. et al. Quantification and characterization of the bystander effect in prostate cancer cells following adenovirus-mediated FasL expression. Cancer Gene Ther 10, 330–339 (2003). https://doi.org/10.1038/sj.cgt.7700576

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