A Heat-Inducible Transgene Expression System for Gene Therapy

Heat-inducible gene expression vectors are useful for hyperthermia-induced cancer gene therapy, because the combination of hyperthermia and gene therapy can considerably improve the therapeutic effects. In the present study, we developed an enhanced heat-inducible transgene expression system in which a heat-shock protein (HSP) promoter and tetracycline-responsive transactivator were combined. When the transactivator plasmid containing the tetracycline-responsive transactivator gene was co-transfected with the reporter gene expression plasmid, a high level of heat-induced gene expression was observed compared with that using the HSP promoter without the transactivator. In vitro evaluation of the therapeutic effect using HeLa cells showed that heat-induced therapeutic gene expression caused cell death in a high percentage of these cells, indicating that this strategy is promising for cancer gene therapy.




References:
[1] W. Walther, J. Wendt, and U. Stein, "Employment of the mdr1 promoter for the chemotherapy-inducible expression of therapeutic genes in cancer gene therapy," Gene Ther, vol. 4, June 1997, pp. 544-552.
[2] D.E. Hallahan, H.J. Mauceri, L.P. Seung, E.J. Dunphy, J.D. Wayne, N.N.
Hanna, A. Toledano, S. Hellman, D.W. Kufe, and R.R. Weichselbaum,
"Spatial and temporal control of gene therapy using ionizing radiation," Nat. Med, vol. 1, August 1995, pp. 786-791
[3] R.V. Blackburn, S.S. Galoforo, P.M. Corry, and Y.J. Lee, "Adenoviral-mediated transfer of a heat-inducible double suicide gene
into prostate carcinoma cells," Cancer Res, vol. 58, April 1998, pp. 1358-1362.
[4] M.W. Dewhirst, L. Prosnitz, D. Thrall, D. Prescott, S. Clegg, C. Charles,
J. MacFall, G. Rosner, T. Samulski, E. Gillette, and S. LaRue, "Hyperthermic treatment of malignant diseases: current status and a view toward the future," Semin Oncol, vol. 24, December 1997, pp. 616-625.
[5] J. van der Zee, "Heating the patient: a promising approach?" Ann. Oncol,
vol. 13, August 2002, pp. 1173-1184.
[6] P. Wust, B. Hildebrandt, G. Sreenivasa, B. Rau, J. Gellermann, H. Riess,
R. Felix, and P.M. Schlag, "Hyperthermia in combined treatment of
cancer," Lancet Oncol, vol. 3, August 2002, pp. 487-497.
[7] W. Walther, and U. Stein, "Heat-responsive gene expression for gene
therapy," Adv. Drug. Deliv. Rev, vol. 61, July 2009, pp. 641-649.
[8] A. Ito, M. Shinkai, H. Honda, and T. Kobayashi, "Heat inducible
TNF-alpha gene therapy combined with hyperthermia using magnetic
nanoparticles as a novel tumor-targeted therapy," Cancer Gene Ther, vol.
8, September 2001, pp. 649-654.
[9] S. Lindquist, "The heat-shock response," Ann. Rev. Biochem, vol. 55,
July 1986, pp. 1151-1191.
[10] T.K. Leung, M.Y. Rajendran, C. Monfries, C. Hall, and L. Lim, "The
human heat-shock protein family. Expression of a novel heat-inducible
HSP70 (HSP70B') and isolation of its cDNA and genomic DNA,"
Biochem. J, vol. 267, April 1990, pp. 125-132.
[11] K. Wada, A. Taniguchi, and T. Okano, "Highly sensitive detection of
cytotoxicity using a modified HSP70B' promoter," Biotechnol. Bioeng,
vol. 97, July 2007, pp. 871-876.
[12] M. Gossen, and H. Bujard, "Tight control of gene expression in
mammalian cells by tetracycline-responsive promoters," Proc. Natl.
Acad. Sci. U.S.A, vol. 89, June 1992, pp. 5547-5551.
[13] M. Kamihira, K. Ono, K. Esaka, K. Nishijima, R. Kigaku, H. Komatsu, T,
Yamashita, K. Kyogoku, and S. Iijima, "High-level expression of
single-chain Fv-Fc fusion protein in serum and egg white of genetically
manipulated chickens by using a retroviral vector," J. Virol, vol. 79,
September 2005, pp. 10864-10874.
[14] A. Ito, F. Matsuoka, H. Honda, and T. Kobayashi, "Antitumor effects of
combined therapy of recombinant heat shock protein 70 and hyperthermia
using magnetic nanoparticles in an experimental subcutaneous murine
melanoma," Cancer Immunol Immunother, vol. 53, January 2004, pp. 26-32.
[15] A. Ito, M. Shinkai, H. Honda, and T. Kobayashi, "Medical application of
functionalized magnetic nanoparticles," J. Biosci. Bioeng, vol. 100, July
2005, pp. 1-11.