Heat shock protein inhibitors increase the efficacy of measles virotherapy

C. Liu, C. Erlichman, C. J. McDonald, J. N. Ingle, P. Zollman, I. Iankov, Stephen J Russell, Evanthia Galanis

Research output: Contribution to journalArticle

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Abstract

Oncolytic measles virus strains have activity against multiple tumor types and are currently in phase I clinical testing. Induction of the heat shock protein 70 (HSP70) constitutes one of the earliest changes in cellular gene expression following infection with RNA viruses including measles virus, and HSP70 upregulation induced by heat shock has been shown to result in increased measles virus cytotoxicity. HSP90 inhibitors such as geldanamycin (GA) or 17-allylaminogeldanamycin result in pharmacologic upregulation of HSP70 and they are currently in clinical testing as cancer therapeutics. We therefore investigated the hypothesis that heat shock protein inhibitors could augment the measles virus-induced cytopathic effect. We tested the combination of a measles virus derivative expressing soluble human carcinoembryonic antigen (MV-CEA) and GA in MDA-MB-231 (breast), SKOV3.IP (ovarian) and TE671 (rhabdomyosarcoma) cancer cell lines. Optimal synergy was accomplished when GA treatment was initiated 6-24h following MV infection. Western immunoblotting confirmed HSP70 upregulation in combination-treated cells. Combination treatment resulted in statistically significant increase in syncytia formation as compared to MV-CEA infection alone. Clonogenic assays demonstrated significant decrease in tumor colony formation in MV-CEA/GA combination-treated cells. In addition there was increase in apoptosis by 4,6-diamidino-2-phenylindole staining. Western immunoblotting for caspase-9, caspase-8, caspase-3 and poly(ADP-ribose) polymerase (PARP) demonstrated increase in cleaved caspase-8 and PARP. The pan-caspase inhibitor Z-VAD-FMK and caspase-8 inhibitor Z-IETD-FMK, but not the caspase-9 inhibitor Z-IEHD-FMK, protected tumor cells from MV-CEA/GA-induced PARP activation, indicating that apoptosis in combination-treated cells occurs mainly via the extrinsic caspase pathway. Treatment of normal cells, such as normal human fibroblasts, however, with the MV-CEA/GA combination, did not result in cytopathic effect, indicating that GA did not alter the MV-CEA specificity for tumor cells. One-step viral growth curves, western immunoblotting for MV-N protein expression, QRT-PCR quantitation of MV-genome copy number and CEA levels showed comparable proliferation of MV-CEA in GA-treated vs -untreated tumor cells. Rho activation assays and western blot for total RhoA, a GTPase associated with the actin cytoskeleton, demonstrated decrease in RhoA activation in combination-treated cells, a change previously shown to be associated with increase in paramyxovirus-induced cell-cell fusion. The enhanced cytopathic effect resulting from measles virus/GA combination supports the translational potential of this approach in the treatment of cancer.

Original languageEnglish (US)
Pages (from-to)1024-1034
Number of pages11
JournalGene Therapy
Volume15
Issue number14
DOIs
StatePublished - Jul 2008

Fingerprint

Measles
Heat-Shock Proteins
Measles virus
HSP70 Heat-Shock Proteins
Caspase Inhibitors
Poly(ADP-ribose) Polymerases
Caspase 8
Neoplasms
Western Blotting
Up-Regulation
Caspase 9
tanespimycin
RNA Virus Infections
Oncolytic Viruses
Apoptosis
geldanamycin
Rhabdomyosarcoma
Cell Fusion
GTP Phosphohydrolases
Carcinoembryonic Antigen

ASJC Scopus subject areas

  • Genetics

Cite this

Liu, C., Erlichman, C., McDonald, C. J., Ingle, J. N., Zollman, P., Iankov, I., ... Galanis, E. (2008). Heat shock protein inhibitors increase the efficacy of measles virotherapy. Gene Therapy, 15(14), 1024-1034. https://doi.org/10.1038/gt.2008.30

Heat shock protein inhibitors increase the efficacy of measles virotherapy. / Liu, C.; Erlichman, C.; McDonald, C. J.; Ingle, J. N.; Zollman, P.; Iankov, I.; Russell, Stephen J; Galanis, Evanthia.

In: Gene Therapy, Vol. 15, No. 14, 07.2008, p. 1024-1034.

Research output: Contribution to journalArticle

Liu, C, Erlichman, C, McDonald, CJ, Ingle, JN, Zollman, P, Iankov, I, Russell, SJ & Galanis, E 2008, 'Heat shock protein inhibitors increase the efficacy of measles virotherapy', Gene Therapy, vol. 15, no. 14, pp. 1024-1034. https://doi.org/10.1038/gt.2008.30
Liu C, Erlichman C, McDonald CJ, Ingle JN, Zollman P, Iankov I et al. Heat shock protein inhibitors increase the efficacy of measles virotherapy. Gene Therapy. 2008 Jul;15(14):1024-1034. https://doi.org/10.1038/gt.2008.30
Liu, C. ; Erlichman, C. ; McDonald, C. J. ; Ingle, J. N. ; Zollman, P. ; Iankov, I. ; Russell, Stephen J ; Galanis, Evanthia. / Heat shock protein inhibitors increase the efficacy of measles virotherapy. In: Gene Therapy. 2008 ; Vol. 15, No. 14. pp. 1024-1034.
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