Induction of Tumor Cell Apoptosis by TRAIL Gene Therapy

  • Thomas S. GriffithEmail author
Part of the Methods in Molecular Biology™ book series (MIMB, volume 542)


Members of the tumor necrosis factor (TNF) superfamily influence a variety of immunological functions, including cellular activation, proliferation, and death, upon interaction with a corresponding superfamily of receptors. Whereas interest in the apoptosis-inducing molecules TNF and Fas ligand has peaked because of their participation in events such as autoimmune disorders, activation-induced cell death, immune privilege, and tumor evasion from the immune system, another death-inducing family member, TNF-related apoptosis-inducing ligand (TRAIL), or Apo-2 ligand, has generated excitement because of its unique ability to induce apoptosis in a wide range of transformed cell lines but not in normal tissues. TRAIL is well tolerated when given to healthy animals, and no observable histological or functional changes have been observed in any of the tissues or organs examined. Moreover, multiple injections of soluble TRAIL into mice beginning the day after tumor implantation can significantly suppress the growth of the tumors, with many animals becoming tumor-free. One potential drawback to these findings is that large amounts of soluble TRAIL may be required to inhibit tumor formation, possibly because of the pharmacokinetic profile of soluble TRAIL that indicates that, after intravenous injection, the majority of the protein is rapidly cleared. Increasing the in vivo half-life of recombinant soluble TRAIL or developing an alternative means of delivery may increase the relative tumoricidal activity of TRAIL such that larger, more established tumors could be eradicated as efficiently as smaller tumors. The information presented here describes the production of an adenoviral vector engineered to carry the complementary DNA (cDNA) for murine TRAIL (hTRAIL).

Key Words

Adenovirus apoptosis gene transfer TRAIL tumor 



I thank Dr. Troy K. Kemp, Tamara Kucaba, and Dr. Rebecca L. VanOosten for their technical assistance; and Maria Scheel, Dalyz Ochoa, and the University of Iowa Gene Transfer Vector Core for virus production. This work was supported by the National Cancer Institute (CA109446).


  1. 1.
    Wiley S.R., Schooley K., Smolak P.J., et al. (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673–682.PubMedCrossRefGoogle Scholar
  2. 2.
    Pitti R.M., Marsters S.A., Ruppert S., Donahue C.J., Moore A., Ashkenazi A. (1996) Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 271:12687–12690.PubMedCrossRefGoogle Scholar
  3. 3.
    Griffith T.S., Chin W.A., Jackson G.C., Lynch D.H., Kubin M.Z. (1998) Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells. J Immunol 161:2833–2840.PubMedGoogle Scholar
  4. 4.
    Walczak H., Miller R.E., Ariail K., et al. (1999) Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 5:157–163.PubMedCrossRefGoogle Scholar
  5. 5.
    Ashkenazi A., Pai R.C., Fong S., et al. (1999) Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 104:155–162.PubMedCrossRefGoogle Scholar
  6. 6.
    Gliniak B., Le T. (1999) Tumor necrosis factor-related apoptosis-inducing ligand's antitumor activity in vivo is enhanced by the chemotherapeutic agent CPT-11. Cancer Res 59:6153–6158.PubMedGoogle Scholar
  7. 7.
    Chinnaiyan A.M., Prasad U., Shankar S., et al. (2000) Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy. Proc Natl Acad Sci USA 97:1754–1759.PubMedCrossRefGoogle Scholar
  8. 8.
    Griffith T.S., Anderson R.D., Davidson B.L., Williams R.D., Ratliff T.L. (2000) Adenoviral-mediated transfer of the TNF-related apoptosis-inducing ligand/Apo-2 ligand gene induces tumor cell apoptosis. J Immunol 165:2886–2894.PubMedGoogle Scholar
  9. 9.
    Griffith T.S., Broghammer E.L. (2001) Suppression of tumor growth following intralesional therapy with TRAIL recombinant adenovirus. Mol Ther 4:257–266.PubMedCrossRefGoogle Scholar
  10. 10.
    Denis L.J. (2000) The role of active treatment in early prostate cancer. Radiother Oncol 57:251–258.PubMedCrossRefGoogle Scholar
  11. 11.
    Stone N.N., Stock R.G. (2000) Prostate brachytherapy in patients with prostate volumes >/= 50 cm(3): dosimetic analysis of implant quality. Int J Radiat Oncol Biol Phys 46:1199–1204.PubMedCrossRefGoogle Scholar
  12. 12.
    Steiner M.S., Gingrich J.R. (2000) Gene therapy for prostate cancer: where are we now? J Urol 164:1121–1136.PubMedCrossRefGoogle Scholar
  13. 13.
    Herman J.R., Adler H.L., Aguilar-Cordova E., et al. (1999) In situ gene therapy for adenocarcinoma of the prostate: a phase I clinical trial. Hum Gene Ther 10:1239–1249.PubMedCrossRefGoogle Scholar
  14. 14.
    Tripathy S.K., Black H.B., Goldwasser E., Leiden J.M. (1996) Immune responses to transgene-encoded proteins limit the stability of gene expression after injection of replication-defective adenovirus vectors. Nat Med 2:545–550.PubMedCrossRefGoogle Scholar
  15. 15.
    Bergelson J.M., Cunningham J.A., Droguett G., et al. (1997) Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5. Science 275:1320–1323.PubMedCrossRefGoogle Scholar
  16. 16.
    Rubin S.A., Rorke L.B. Adenoviral vaccines. In: Plotkin M, ed. Vaccines. Philadelphia: W.B. Saunders; 1990:492–512.Google Scholar
  17. 17.
    Akli S., Caillaud C., Vigne E., et al. (1993) Transfer of a foreign gene into the brain using adenovirus vectors. Nat Genet 3:224–228.PubMedCrossRefGoogle Scholar
  18. 18.
    Baratin M., Ziol M., Romieu R., et al. (2001) Regression of primary hepatocarcinoma in cancer-prone transgenic mice by local interferon-gamma delivery is associated with macrophages recruitment and nitric oxide production. Cancer Gene Ther 8:193–202.PubMedCrossRefGoogle Scholar
  19. 19.
    Cordier L., Duffour M.T., Sabourin J.C., et al. (1995) Complete recovery of mice from a pre-established tumor by direct intratumoral delivery of an adenovirus vector harboring the murine IL-2 gene. Gene Ther 2:16–21.PubMedGoogle Scholar
  20. 20.
    Le Gal La Salle G., Robert J.J., Berrard S., et al. (1993) An adenovirus vector for gene transfer into neurons and glia in the brain. Science 259:988–990.PubMedCrossRefGoogle Scholar
  21. 21.
    Rosenfeld M.A., Yoshimura K., Trapnell B.C., et al. (1992) In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium. Cell 68:143–155.PubMedCrossRefGoogle Scholar
  22. 22.
    Timme T.L., Hall S.J., Barrios R., Woo S.L., Aguilar-Cordova E., Thompson T.C. (1998) Local inflammatory response and vector spread after direct intraprostatic injection of a recombinant adenovirus containing the herpes simplex virus thymidine kinase gene and ganciclovir therapy in mice. Cancer Gene Ther 5:74–82.PubMedGoogle Scholar
  23. 23.
    Murphy G.P., Hrushesky W.J. (1973) A murine renal cell carcinoma. J Natl Cancer Inst 50:1013–1025.PubMedGoogle Scholar
  24. 24.
    Kayagaki N., Yamaguchi N., Nakayama M., et al. (1999) Expression and function of TNF-related apoptosis-inducing ligand on murine activated NK cells. J Immunol 163:1906–1913.PubMedGoogle Scholar
  25. 25.
    Anderson R.D., Haskell R.E., Xia H., Roessler B.J., Davidson B.L. (2000) A simple method for the rapid generation of recombinant adenovirus vectors. Gene Ther 7:1034–1038.PubMedCrossRefGoogle Scholar
  26. 26.
    Flick D.A., Gifford G.E. (1984) Comparison of in vitro cell cytotoxic assays for tumor necrosis factor. J Immunol Methods 68:167–175.PubMedCrossRefGoogle Scholar
  27. 27.
    Martin S.J., Reutelingsperger C.P., McGahon A.J., et al. (1995) Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 182:1545–1556.PubMedCrossRefGoogle Scholar
  28. 28.
    Hemminki A., Alvarez R.D. (2002) Adenoviruses in oncology: a viable option? BioDrugs 16:77–87.PubMedCrossRefGoogle Scholar
  29. 29.
    Okegawa T., Li Y., Pong R.C., Bergelson J.M., Zhou J., Hsieh J.T. (2000) The dual impact of coxsackie and adenovirus receptor expression on human prostate cancer gene therapy. Cancer Res 60:5031–5036.PubMedGoogle Scholar
  30. 30.
    Eastham J.A., Hall S.J., Sehgal I., et al. (1995) In vivo gene therapy with p53 or p21 adenovirus for prostate cancer. Cancer Res 55:5151–5155.PubMedGoogle Scholar
  31. 31.
    Hall S.J., Mutchnik S.E., Yang G., et al. (1999) Cooperative therapeutic effects of androgen ablation and adenovirus-mediated herpes simplex virus thymidine kinase gene and ganciclovir therapy in experimental prostate cancer. Cancer Gene Ther 6:54–63.PubMedCrossRefGoogle Scholar
  32. 32.
    Nasu Y., Bangma C.H., Hull G.W., et al. (1999) Adenovirus-mediated interleukin-12 gene therapy for prostate cancer: suppression of orthotopic tumor growth and pre-established lung metastases in an orthotopic model. Gene Ther 6:338–349.PubMedCrossRefGoogle Scholar
  33. 33.
    Anello R., Cohen S., Atkinson G., Hall S.J. (2000) Adenovirus mediated cytosine deaminase gene transduction and 5-fluorocytosine therapy sensitizes mouse prostate cancer cells to irradiation. J Urol 164:2173–2177.PubMedCrossRefGoogle Scholar
  34. 34.
    Shariat S.F., Desai S., Song W., et al. (2001) Adenovirus-mediated transfer of inducible caspases: a novel “death switch” gene therapeutic approach to prostate cancer. Cancer Res 61:2562–2571.PubMedGoogle Scholar
  35. 35.
    Cao G., Su J., Lu W., et al. (2001) Adenovirus-mediated interferon-beta gene therapy suppresses growth and metastasis of human prostate cancer in nude mice. Cancer Gene Ther 8:497–505.PubMedCrossRefGoogle Scholar
  36. 36.
    Katner A.L., Hoang Q.B., Gootam P., et al. (2002) Induction of cell cycle arrest and apoptosis in human prostate carcinoma cells by a recombinant adenovirus expressing p27(Kip1). Prostate 53:77–87.PubMedCrossRefGoogle Scholar
  37. 37.
    Flynn V., Jr., Ramanitharan A., Moparty K., et al. (2003) Adenovirus-mediated inhibition of NF-kappaB confers chemo-sensitization and apoptosis in prostate cancer cells. Int J Oncol 23:317–323.PubMedGoogle Scholar
  38. 38.
    Voelkel-Johnson C., King D.L., Norris J.S. (2002) Resistance of prostate cancer cells to soluble TNF-related apoptosis-inducing ligand (TRAIL/Apo2L) can be overcome by doxorubicin or adenoviral delivery of full-length TRAIL. Cancer Gene Ther 9:164–172.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Department of Urology and the Interdisciplinary Graduate Program in ImmunologyUniversity of IowaIowa CityUSA

Personalised recommendations