Abstract
Reovirus shows considerable potential as an oncolytic agent for Ras-activated tumors and is currently in clinical trials. Here we ask whether such tumor cell lines can acquire resistance to reoviral oncolysis. We challenged human HT1080 fibrosarcoma cells that carry a Ras mutation by prolonged exposure to reovirus, thereby yielding highly virus-resistant HTR1 cells. These cells are persistently infected with reovirus, exhibit high Ras activity and retain the original Ras gene mutation, showing that resistance to reovirus can be displayed in cells with active Ras. The HTR1 cells also exhibit reduced cellular cathepsin B activity, which normally contributes to viral entry and activation. Persistently infected HTR1 cells were not tumorigenic in vivo, whereas immunologically cured virus-free HTR1 cells were highly tumorigenic. Thus, acquisition of resistance to reovirus may constrain therapeutic strategies. To determine whether reoviral resistance is associated with a general reduction in apoptotic potential, we challenged the HTR1 cells with apoptotic inducers and E1B-defective adenovirus, resulting in significant apoptosis and cell death following both approaches. Therefore, even if resistance to reoviral oncolysis should arise in tumor cells in vivo, other therapeutic strategies may nevertheless remain effective.
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References
Ahmed R, Canning WM, Kauffman RS, Sharpe AH, Hallum JV, Fields BN . (1981). Role of the host cell in persistent viral infection: coevolution of L cells and reovirus during persistent infection. Cell 25: 325–332.
Baer GS, Ebert DH, Chung CJ, Erickson AH, Dermody TS . (1999). Mutant cells selected during persistent reovirus infection do not express mature cathepsin L and do not support reovirus disassembly. J Virol 73: 9532–9543.
Barrett AJ, Kirschke H . (1981). Cathepsin B, cathepsin H, and cathepsin L. Methods Enzymol 80: 535–561.
Brown R, Marshall CJ, Pennie SG, Hall A . (1984). Mechanism of activation of an N-ras gene in the human fibrosarcoma cell line HT1080. EMBO J 3: 1321–1326.
Buttle DJ, Murata M, Knight CG, Barrett AJ . (1992). CA074 methyl ester: a proinhibitor for intracellular cathepsin B. Arch Biochem Biophys 299: 377–380.
Clarke P, Meintzer SM, Spalding AC, Johnson GL, Tyler KL . (2001). Caspase 8-dependent sensitization of cancer cells to TRAIL-induced apoptosis following reovirus-infection. Oncogene 20: 6910–6919.
Coffey MC, Strong JE, Forsyth PA, Lee PW . (1998). Reovirus therapy of tumors with activated Ras pathway. Science 282: 1332–1334.
Connolly JL, Dermody TS . (2002). Virion disassembly is required for apoptosis induced by reovirus. J Virol 76: 1632–1641.
Danis C, Mabrouk T, Faure M, Lemay G . (1997). Interferon has no protective effect during acute or persistent reovirus infection of mouse SC1 fibroblasts. Virus Res 51: 139–149.
De Rooij J, Bos JL . (1997). Minimal Ras-binding domain of Raf1 can be used as an activation-specific probe for Ras. Oncogene 14: 623–625.
Dermody TS . (1998). Molecular mechanisms of persistent infection by reovirus. Curr Top Microbiol Immunol 233: 1–22.
Dermody TS, Chappell JD, Hofler JG, Kramp W, Tyler KL . (1995). Eradication of persistent reovirus infection from a B-cell hybridoma. Virology 212: 272–276.
Duncan MR, Stanish SM, Cox DC . (1978). Differential sensitivity of normal and transformed human cells to reovirus infection. J Virol 28: 444–449.
Duursma AM, Agami R . (2003). Ras interference as cancer therapy. Semin Cancer Biol 13: 267–273.
Ebert DH, Kopecky-Bromberg SA, Dermody TS . (2004). Cathepsin B is inhibited in mutant cells selected during persistent reovirus infection. J Biol Chem 279: 3837–3851.
Hashiro G, Loh PC, Yau JT . (1977). The preferential cytotoxicity of reovirus for certain transformed cell lines. Arch Virol 54: 307–315.
Kominsky DJ, Bickel RJ, Tyler KL . (2002). Reovirus-induced apoptosis requires both death receptor- and mitochondrial-mediated caspase-dependent pathways of cell death. Cell Death Differ 9: 926–933.
Langland JO, Cameron JM, Heck MC, Jancovich JK, Jacobs BL . (2006). Inhibition of PKR by RNA and DNA viruses. Virus Res 119: 100–110.
Loken SD, Norman K, Hirasawa K, Nodwell M, Lester WM, Demetrick DJ . (2004). Morbidity in immunosuppressed (SCID/NOD) mice treated with reovirus (dearing 3) as an anti-cancer biotherapeutic. Cancer Biol Ther 3: 734–738.
Meusel TR, Imani F . (2003). Viral induction of inflammatory cytokines in human epithelial cells follows a p38 mitogen-activated protein kinase-dependent but NF-kappa B-independent pathway. J Immunol 171: 3768–3774.
Mosmann T . (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55–63.
Nagata L, Masri SA, Mah DC, Lee PW . (1984). Molecular cloning and sequencing of the reovirus (serotype 3) S1 gene which encodes the viral cell attachment protein sigma 1. Nucleic Acids Res 12: 8699–8710.
Norman KL, Lee PW . (2005). Not all viruses are bad guys: the case for reovirus in cancer therapy. Drug Discov Today 10: 847–855.
Paterson H, Reeves B, Brown R, Hall A, Furth M, Bos J et al. (1987). Activated N-ras controls the transformed phenotype of HT1080 human fibrosarcoma cells. Cell 51: 803–812.
Samuel CE . (1998). Reoviruses and the interferon system. Curr Top Microbiol Immunol 233: 125–145.
Schotte P, Schauvliege R, Janssens S, Beyaert R . (2001). The cathepsin B inhibitor z-FA.fmk inhibits cytokine production in macrophages stimulated by lipopolysaccharide. J Biol Chem 276: 21153–21157.
Shiratsuchi A, Kaido M, Takizawa T, Nakanishi Y . (2000). Phosphatidylserine-mediated phagocytosis of influenza A virus-infected cells by mouse peritoneal macrophages. J Virol 74: 9240–9244.
Smith RE, Zweerink HJ, Joklik WK . (1969). Polypeptide components of virions, top component and cores of reovirus type 3. Virology 39: 791–810.
Steele TA, Cox DC . (1996). Elimination of EL-4 and L1210 murine tumors by 1,3-bis (2-chloroethyl)-1-nitrosourea requires an intact immune response. Proc Soc Exp Biol Med 212: 63–68.
Steele TA, Hauser CC . (2005). The role of interferon-alpha in a successful murine tumor therapy. Exp Biol Med (Maywood) 230: 487–493.
Stewart MJ, Smoak K, Blum MA, Sherry B . (2005). Basal and reovirus-induced beta interferon (IFN-beta) and IFN-beta-stimulated gene expression are cell type specific in the cardiac protective response. J Virol 79: 2979–2987.
Stoeckel J, Hay JG . (2006). Drug evaluation: Reolysin – wild-type reovirus as a cancer therapeutic. Curr Opin Mol Ther 8: 249–260.
Strong JE, Coffey MC, Tang D, Sabinin P, Lee PW . (1998). The molecular basis of viral oncolysis: usurpation of the Ras signaling pathway by reovirus. EMBO J 17: 3351–3362.
Taber R, Alexander V, Whitford W . (1976). Persistent reovirus infection of CHO cells resulting in virus resistance. J Virol 17: 513–524.
Teodoro JG, Halliday T, Whalen SG, Takayesu D, Graham FL, Branton PE . (1994). Phosphorylation at the carboxy terminus of the 55-kilodalton adenovirus type 5 E1B protein regulates transforming activity. J Virol 68: 776–786.
Tyler KL . (2001). Mammalian reoviruses. In: Knipe DM, Howley PM (eds). Lippincott Fields Virology. Williams & Wilkens: Philadelphia, pp 1729–1745.
Wong H, Anderson WD, Cheng T, Riabowol KT . (1994). Monitoring mRNA expression by polymerase chain reaction: the ‘primer-dropping’ method. Anal Biochem 223: 251–258.
Yeung MC, Chang DL, Camantigue RE, Lau AS . (1999). Inhibitory role of the host apoptogenic gene PKR in the establishment of persistent infection by encephalomyocarditis virus in U937 cells. Proc Natl Acad Sci USA 96: 11860–11865.
Acknowledgements
Portions of this work were completed as part of a PhD thesis by MK. This study was supported by research grants to RNJ from the Alberta Cancer Board and the Canadian Institutes for Health Research.
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Kim, M., Egan, C., Alain, T. et al. Acquired resistance to reoviral oncolysis in Ras-transformed fibrosarcoma cells. Oncogene 26, 4124–4134 (2007). https://doi.org/10.1038/sj.onc.1210189
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DOI: https://doi.org/10.1038/sj.onc.1210189
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