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Mechanisms involved in development of resistance to adenovirus-mediated proapoptotic gene therapy in DLD1 human colon cancer cell line

Gene Therapy volume 9, pages 1262–1270 (2002)Cite this article

Abstract

To evaluate resistance that develops in cancer cells during treatment with adenoviral vectors expressing proapoptotic genes, we repeatedly treated the human colon cancer cell line DLD1 with adenoviral vectors expressing the human Bax gene and the human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene. DLD1 cells resistant to the Bax- or TRAIL-expressing adenoviral vectors were then selected and designated as DLD1/Bax-R or DLD1/TRAIL-R cells, respectively. Further study showed that resistance in DLD1/Bax-R cells was caused by resistance to adenoviral infection, which can be overcome by dose escalation of the adenoviral vectors. However, resistance in DLD1/TRAIL-R cells was caused by resistance to the TRAIL gene. Therefore, different mechanisms are involved in the development of resistance during adenovirus-mediated proapoptotic gene therapy. A survey of molecules involved in TRAIL- or Bax-mediated apoptotic pathways showed no significant change in expression of death receptors, death decoy receptors; FLIP; Bcl-2; Bcl-xS; Bax; Bak; XIAP or caspase-2, -7, -8, or -9 in either DLD1/Bax-R or DLD1/TRAIL-R cells. Bcl-xL expression detected in both mRNA and protein level assays was three times higher in DLD1/TRAIL-R cells than in parental or DLD1/Bax-R cells. However, transfection of DLD1 cells with the Bcl-xL gene showed that overexpression of Bcl-xL is not sufficient for the resistance. Moreover, DLD1/Bax-R cells were sensitive to adenoviral vectors that expressed the TRAIL gene, but resistant to adenoviral vectors that expressed the Bak gene. In contrast, DLD1/TRAIL-R cells were sensitive to adenoviral vectors that expressed either Bax or Bak gene. Thus, alternative application of adenoviral vectors that expressed proapoptotic genes in different pathways or different cell killing models may delay or prevent development of resistance in adenovirus-mediated proapoptotic gene therapy.

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References

  1. Kagawa S et al. Antitumor effect of adenovirus-mediated Bax gene transfer on p53-sensitive and p53-resistant cancer lines Cancer Res 2000 60: 1157–1161

    CAS  PubMed  Google Scholar 

  2. Gu J et al. Tumor-specific transgene expression from the human telomerase reverse transcriptase promoter enables targeting of the therapeutic effects of the Bax gene to cancers Cancer Res 2000 60: 5359–5364

    CAS  PubMed  Google Scholar 

  3. Li X et al. Adenovirus-mediated Bax overexpression for the induction of therapeutic apoptosis in prostate cancer Cancer Res 2001 61: 186–191

    CAS  PubMed  Google Scholar 

  4. Tai YT, Strobel T, Kufe D, Cannistra SA . In vivo cytotoxicity of ovarian cancer cells through tumor-selective expression of the BAX gene Cancer Res 1999 59: 2121–2126

    CAS  PubMed  Google Scholar 

  5. Kagawa S et al. Antitumor activity and bystander effects of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene Cancer Res 2001 61: 3330–3338

    CAS  PubMed  Google Scholar 

  6. Griffith TS et al. Adenoviral-mediated transfer of the TNF-related apoptosis-inducing ligand/Apo-2 ligand gene induces tumor cell apoptosis J Immunol 2000 165: 2886–2894

    Article  CAS  PubMed  Google Scholar 

  7. Jurgensmeier JM et al. Bax directly induces release of cytochrome c from isolated mitochondria Proc Natl Acad Sci USA 1998 95: 4997–5002

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Pastorino JG et al. The overexpression of Bax produces cell death upon induction of the mitochondrial permeability transition J Biol Chem 1998 273: 7770–7775

    Article  CAS  PubMed  Google Scholar 

  9. Liu X et al. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c Cell 1996 86: 147–157

    Article  CAS  PubMed  Google Scholar 

  10. Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD . The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis Science 1997 275: 1132–1136

    Article  CAS  PubMed  Google Scholar 

  11. Oltvai ZN, Milliman CL, Korsmeyer SJ . Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death Cell 1993 74: 609–619

    Article  CAS  PubMed  Google Scholar 

  12. Pan G et al. The receptor for the cytotoxic ligand TRAIL Science 1991 276: 111–113

    Article  Google Scholar 

  13. Walczak H et al. TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL EMBO J 1997 16: 5386–5397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Eggert A et al. Resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in neuroblastoma cells correlates with a loss of caspase-8 expression Cancer Res 2001 61: 1314–1319

    CAS  PubMed  Google Scholar 

  15. Zhang XD et al. Mechanisms of resistance of normal cells to TRAIL induced apoptosis vary between different cell types FEBS Lett 2000 482: 193–199

    Article  CAS  PubMed  Google Scholar 

  16. Zhang XD et al. Tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis of human melanoma is regulated by smac/DIABLO release from mitochondria Cancer Res 2001 61: 7339–7348

    CAS  PubMed  Google Scholar 

  17. Pan G et al. An antagonist decoy receptor and a death domain-containing receptor for TRAIL Science 1997 277: 815–818

    Article  CAS  PubMed  Google Scholar 

  18. Sheridan JP et al. Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors Science 1997 277: 818–821

    Article  CAS  PubMed  Google Scholar 

  19. Degli-Esposti MA et al. The novel receptor TRAIL-R4 induces NF-kappaB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain Immunity 1997 7: 813–820

    Article  CAS  PubMed  Google Scholar 

  20. Emery JG et al. Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL J Biol Chem 1998 273: 14363–14367

    Article  CAS  PubMed  Google Scholar 

  21. Favrot M, Coll JL, Louis N, Negoescu A . Cell death and cancer: replacement of apoptotic genes and inactivation of death suppressor genes in therapy Gene Therapy 1998 5: 728–739

    Article  CAS  PubMed  Google Scholar 

  22. Kagawa S et al. A binary adenoviral vector for expressing high levels of the proapoptotic gene bax Gene Therapy 2000 7: 75–79

    Article  CAS  PubMed  Google Scholar 

  23. Bergelson JM et al. Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5 Science 1997 275: 1320–1323

    Article  CAS  PubMed  Google Scholar 

  24. Davison E et al. Integrin alpha5beta1-mediated adenovirus infection is enhanced by the integrin-activating antibody TS2/16 J Virol 1997 71: 6204–6207

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Goldman MJ, Wilson JM . Expression of alpha v beta 5 integrin is necessary for efficient adenovirus-mediated gene transfer in the human airway J Virol 1995 69: 5951–5958

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Pearson AS et al. Factors limiting adenovirus-mediated gene transfer into human lung and pancreatic cancer cell lines Clin Cancer Res 1999 5: 4208–4213

    CAS  PubMed  Google Scholar 

  27. Wickham TJ, Mathias P, Cheresh DA, Nemerow GR . Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment Cell 1993 73: 309–319

    Article  CAS  PubMed  Google Scholar 

  28. Wickham TJ, Roelvink PW, Brough DE, Kovesdi I . Adenovirus targeted to heparan-containing receptors increases its gene delivery efficiency to multiple cell types Nature Biotech 1996 14: 1570–1573

    Article  CAS  Google Scholar 

  29. Dmitriev I et al. An adenovirus vector with genetically modified fibers demonstrates expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent cell entry mechanism J Virol 1998 72: 9706–9713

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Wickham TJ, Carrion ME, Kovesdi I . Targeting of adenovirus penton base to new receptors through replacement of its RGD motif with other receptor-specific peptide motifs Gene Therapy 1995 2: 750–756

    CAS  PubMed  Google Scholar 

  31. Hinz S et al. Bcl-XL protects pancreatic adenocarcinoma cells against CD95- and TRAIL-receptor-mediated apoptosis Oncogene 2000 19: 5477–5486

    Article  CAS  PubMed  Google Scholar 

  32. Walczak H, Bouchon A, Stahl H, Krammer PH . Tumor necrosis factor-related apoptosis-inducing ligand retains its apoptosis-inducing capacity on Bcl-2- or Bcl-xL-overexpressing chemotherapy-resistant tumor cells Cancer Res 2000 60: 3051–3057

    CAS  PubMed  Google Scholar 

  33. Fang B, Ji L, Bouvet M, Roth JA . Evaluation of GAL4/TATA in vivo. Induction of transgene expression by adenovirally mediated gene codelivery J Biol Chem 1998 273: 4972–4975

    Article  CAS  PubMed  Google Scholar 

  34. Koch P et al. Augmenting transgene expression from carcinoembryonic antigen (CEA) promoter via a GAL4 gene regulatory system Mol Ther J Am Soc Gene Ther 2001 3: 278–283

    Article  CAS  Google Scholar 

  35. Fang B, Xu B, Koch P, Roth JA . Intercellular trafficking of VP22-GFP fusion proteins is not observed in cultured mammalian cells Gene Therapy 1998 5: 1420–1424

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported in part by a research project grant from the American Cancer Society (RPG-00-274-01-MGO to BF); an Institutional Start-Up Fund (to BF); a grant from the WM Keck Center for Cancer Gene Therapy, The University of Texas MD Anderson Cancer Center (BF); an NIH project grant (P01 CA78778-01A1, to JAR); and an NIH Core Grant (CA16672). JG is an MD Anderson Cancer Center Odyssey Program Fellow supported by the Kimberly-Clark Endowment for New and Innovative Research. We thank Allan Prejusa and Trupti Mehta for adenovirus propagation and quality control and Kate O Suilleabhain for editorial review.

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  1. Department of Thoracic and Cardiovascular Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    L Zhang, J Gu, T Lin, X Huang, J A Roth & B Fang

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  1. L Zhang
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  2. J Gu
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Zhang, L., Gu, J., Lin, T. et al. Mechanisms involved in development of resistance to adenovirus-mediated proapoptotic gene therapy in DLD1 human colon cancer cell line. Gene Ther 9, 1262–1270 (2002). https://doi.org/10.1038/sj.gt.3301797

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