Abstract
The E2F-1 promoter has been used to confer tumor-selective E1A expression in oncolytic adenoviruses. Tumor specificity is mainly conferred by a unique structure of E2F-responsive sites organized in palindromes. Binding of the E2F–pRb complex to these palindromes results in repression of transcription in normal cells. Owing to deregulation of the Rb/p16 pathway in tumor cells, binding of free E2F activates transcription and initiates an autoactivation loop involving E1A and E4-6/7. ICOVIR-7 is a new oncolytic adenovirus designed to increase the E2F dependency of E1A gene expression. It incorporates additional palindromes of E2F-responsive sites in an insulated E2F-1 promoter controlling E1A-Δ24. The E2F palindromes inhibited replication in normal cells, resulting in a low systemic toxicity at high doses in immunocompetent mice. The Δ24 deletion avoids a loop of E2F-mediated self-activation in nontumor cells. Importantly, the additional E2F-binding hairpins boost the positive feedback loop on the basis of E1A-mediated transcriptional regulation of E4-6/7 turned on in cancer cells and increased antitumoral potency as shown in murine subcutaneous xenograft models treated by intravenous injection. These results suggest that the unique genetic combination featured in ICOVIR-7 may be promising for treating disseminated neoplasias.
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References
Bauzon M, Hermiston TW . Exploiting diversity: genetic approaches to creating highly potent and efficacious oncolytic viruses. Curr Opin Mol Ther 2008; 10: 350–355.
Rodriguez R, Schuur ER, Lim HY, Henderson GA, Simons JW, Henderson DR . Prostate attenuated replication competent adenovirus (ARCA) CN706: a selective cytotoxic for prostate-specific antigen-positive prostate cancer cells. Cancer Res 1997; 57: 2559–2563.
Fuerer C, Iggo R . Adenoviruses with Tcf binding sites in multiple early promoters show enhanced selectivity for tumour cells with constitutive activation of the wnt signalling pathway. Gene Therapy 2002; 9: 270–281.
Sherr CJ, McCormick F . The RB and p53 pathways in cancer. Cancer Cell 2002; 2: 103–112.
Black AR, Azizkhan-Clifford J . Regulation of E2F: a family of transcription factors involved in proliferation control. Gene 1999; 237: 281–302.
Johnson DG, Degregori J . Putting the oncogenic and tumor suppressive activities of E2F into context. Curr Mol Med 2006; 6: 731–738.
Neuman E, Flemington EK, Sellers WR, Kaelin Jr WG . Transcription of the E2F-1 gene is rendered cell cycle dependent by E2F DNA-binding sites within its promoter. Mol Cell Biol 1994; 14: 6607–6615.
Neuman E, Flemington EK, Sellers WR, Kaelin Jr WG . Transcription of the E2F-1 gene is rendered cell cycle dependent by E2F DNA-binding sites within its promoter. Mol Cell Biol 1995; 15: 4660.
Kovesdi I, Reichel R, Nevins JR . Identification of a cellular transcription factor involved in E1A trans-activation. Cell 1986; 45: 219–228.
Cress WD, Nevins JR . Use of the E2F transcription factor by DNA tumor virus regulatory proteins. Curr Top Microbiol Immunol 1996; 208: 63–78.
Schaley J, O’Connor RJ, Taylor LJ, Bar-Sagi D, Hearing P . Induction of the cellular E2F-1 promoter by the adenovirus E4-6/7 protein. J Virol 2000; 74: 2084–2093.
Fueyo J, Gomez-Manzano C, Alemany R, Lee PS, McDonnell TJ, Mitlianga P et al. A mutant oncolytic adenovirus targeting the Rb pathway produces anti-glioma effect in vivo. Oncogene 2000; 19: 2–12.
Heise C, Hermiston T, Johnson L, Brooks G, Sampson-Johannes A, Williams A et al. An adenovirus E1A mutant that demonstrates potent and selective systemic anti-tumoral efficacy. Nat Med 2000; 6: 1134–1139.
Jakubczak JL, Ryan P, Gorziglia M, Clarke L, Hawkins LK, Hay C et al. An oncolytic adenovirus selective for retinoblastoma tumor suppressor protein pathway-defective tumors: dependence on E1A, the E2F-1 promoter, and viral replication for selectivity and efficacy. Cancer Res 2003; 63: 1490–1499.
Alonso MM, Cascallo M, Gomez-Manzano C, Jiang H, Bekele BN, Perez-Gimenez A et al. ICOVIR-5 shows E2F1 addiction and potent antiglioma effect in vivo. Cancer Res 2007; 67: 8255–8263.
Alonso MM, Gomez-Manzano C, Jiang H, Bekele NB, Piao Y, Yung WK et al. Combination of the oncolytic adenovirus ICOVIR-5 with chemotherapy provides enhanced anti-glioma effect in vivo. Cancer Gene Ther 2007; 14: 756–761.
Cascallo M, Alonso MM, Rojas JJ, Perez-Gimenez A, Fueyo J, Alemany R . Systemic toxicity-efficacy profile of ICOVIR-5, a potent and selective oncolytic adenovirus based on the pRB pathway. Mol Ther 2007; 15: 1607–1615.
Engler H, Machemer T, Philopena J, Wen SF, Quijano E, Ramachandra M et al. Acute hepatotoxicity of oncolytic adenoviruses in mouse models is associated with expression of wild-type E1a and induction of TNF-alpha. Virology 2004; 328: 52–61.
Kirby TO, Rivera A, Rein D, Wang M, Ulasov I, Breidenbach M et al. A novel ex vivo model system for evaluation of conditionally replicative adenoviruses therapeutic efficacy and toxicity. Clin Cancer Res 2004; 10: 8697–8703.
Morral N, O’Neal WK, Rice K, Leland MM, Piedra PA, Aguilar-Cordova E et al. Lethal toxicity, severe endothelial injury, and a threshold effect with high doses of an adenoviral vector in baboons. Hum Gene Ther 2002; 13: 143–154.
Duncan SJ, Gordon FC, Gregory DW, McPhie JL, Postlethwaite R, White R et al. Infection of mouse liver by human adenovirus type 5. J Gen Virol 1978; 40: 45–61.
Nemunaitis J, Cunningham C, Buchanan A, Blackburn A, Edelman G, Maples P et al. Intravenous infusion of a replication-selective adenovirus (ONYX-015) in cancer patients: safety, feasibility and biological activity. Gene Therapy 2001; 8: 746–759.
Aghi M, Martuza RL . Oncolytic viral therapies—the clinical experience. Oncogene 2005; 24: 7802–7816.
Cichon G, Schmidt HH, Benhidjeb T, Loser P, Ziemer S, Haas R et al. Intravenous administration of recombinant adenoviruses causes thrombocytopenia, anemia and erythroblastosis in rabbits. J Gene Med 1999; 1: 360–371.
Rossini M . The role of adenovirus early region 1A in the regulation of early regions 2A and 1B expression. Virology 1983; 131: 49–58.
Weeks DL, Jones NC . E1A control of gene expression is mediated by sequences 5′ to the transcriptional starts of the early viral genes. Mol Cell Biol 1983; 3: 1222–1234.
Majem M, Cascallo M, Bayo-Puxan N, Mesia R, Germa JR, Alemany R . Control of E1A under an E2F-1 promoter insulated with the myotonic dystrophy locus insulator reduces the toxicity of oncolytic adenovirus Ad-Delta24RGD. Cancer Gene Ther 2006; 13: 696–705.
Ricciardelli C, Russell DL, Ween MP, Mayne K, Suwiwat S, Byers S et al. Formation of hyaluronan- and versican-rich pericellular matrix by prostate cancer cells promotes cell motility. J Biol Chem 2007; 282: 10814–10825.
Draffin JE, McFarlane S, Hill A, Johnston PG, Waugh DJ . CD44 potentiates the adherence of metastatic prostate and breast cancer cells to bone marrow endothelial cells. Cancer Res 2004; 64: 5702–5711.
Habib N, Salama H, Abd El Latif Abu Median A, Isac Anis I, Abd Al Aziz RA, Sarraf C et al. Clinical trial of E1B-deleted adenovirus (dl1520) gene therapy for hepatocellular carcinoma. Cancer Gene Ther 2002; 9: 254–259.
Small EJ, Carducci MA, Burke JM, Rodriguez R, Fong L, van Ummersen L et al. A phase I trial of intravenous CG7870, a replication-selective, prostate-specific antigen-targeted oncolytic adenovirus, for the treatment of hormone-refractory, metastatic prostate cancer. Mol Ther 2006; 14: 107–117.
Hamid O, Varterasian ML, Wadler S, Hecht JR, Benson 3rd A, Galanis E et al. Phase II trial of intravenous CI-1042 in patients with metastatic colorectal cancer. J Clin Oncol 2003; 21: 1498–1504.
Dyson N . The regulation of E2F by pRB-family proteins. Genes Dev 1998; 12: 2245–2262.
Suzuki K, Fueyo J, Krasnykh V, Reynolds PN, Curiel DT, Alemany R . A conditionally replicative adenovirus with enhanced infectivity shows improved oncolytic potency. Clin Cancer Res 2001; 7: 120–126.
Tsukuda K, Wiewrodt R, Molnar-Kimber K, Jovanovic VP, Amin KM . An E2F-responsive replication-selective adenovirus targeted to the defective cell cycle in cancer cells: potent antitumoral efficacy but no toxicity to normal cell. Cancer Res 2002; 62: 3438–3447.
Ryan PC, Jakubczak JL, Stewart DA, Hawkins LK, Cheng C, Clarke LM et al. Antitumor efficacy and tumor-selective replication with a single intravenous injection of OAS403, an oncolytic adenovirus dependent on two prevalent alterations in human cancer. Cancer Gene Ther 2004; 11: 555–569.
Johnson L, Shen A, Boyle L, Kunich J, Pandey K, Lemmon M et al. Selectively replicating adenoviruses targeting deregulated E2F activity are potent, systemic antitumor agents. Cancer Cell 2002; 1: 325–337.
Neill SD, Hemstrom C, Virtanen A, Nevins JR . An adenovirus E4 gene product trans-activates E2 transcription and stimulates stable E2F binding through a direct association with E2F. Proc Natl Acad Sci USA 1990; 87: 2008–2012.
Nettelbeck DM, Rivera AA, Balague C, Alemany R, Curiel DT . Novel oncolytic adenoviruses targeted to melanoma: specific viral replication and cytolysis by expression of E1A mutants from the tyrosinase enhancer/promoter. Cancer Res 2002; 62: 4663–4670.
Hitt MM, Graham FL . Adenovirus E1A under the control of heterologous promoters: wide variation in E1A expression levels has little effect on virus replication. Virology 1990; 179: 667–678.
Liu Q, Zaiss AK, Colarusso P, Patel K, Haljan G, Wickham TJ et al. The role of capsid-endothelial interactions in the innate immune response to adenovirus vectors. Hum Gene Ther 2003; 14: 627–643.
Alemany R . Cancer selective adenoviruses. Mol Aspects Med 2007; 28: 42–58.
Cheng J, Sauthoff H, Huang Y, Kutler DI, Bajwa S, Rom WN et al. Human matrix metalloproteinase-8 gene delivery increases the oncolytic activity of a replicating adenovirus. Mol Ther 2007; 15: 1982–1990.
Ganesh S, Gonzalez-Edick M, Gibbons D, Van Roey M, Jooss K . Intratumoral coadministration of hyaluronidase enzyme and oncolytic adenoviruses enhances virus potency in metastatic tumor models. Clin Cancer Res 2008; 14: 3933–3941.
Kim JH, Lee YS, Kim H, Huang JH, Yoon AR, Yun CO . Relaxin expression from tumor-targeting adenoviruses and its intratumoral spread, apoptosis induction, and efficacy. J Natl Cancer Inst 2006; 98: 1482–1493.
Fuerer C, Iggo R . 5-Fluorocytosine increases the toxicity of Wnt-targeting replicating adenoviruses that express cytosine deaminase as a late gene. Gene Therapy 2004; 11: 142–151.
Lukashev AN, Fuerer C, Chen MJ, Searle P, Iggo R . Late expression of nitroreductase in an oncolytic adenovirus sensitizes colon cancer cells to the prodrug CB1954. Hum Gene Ther 2005; 16: 1473–1483.
Cascante A, Abate-Daga D, Garcia-Rodriguez L, Gonzalez JR, Alemany R, Fillat C . GCV modulates the antitumoural efficacy of a replicative adenovirus expressing the Tat8-TK as a late gene in a pancreatic tumour model. Gene Therapy 2007; 14: 1471–1480.
Guedan S, Gros A, Cascallo M, Vile R, Mercade E, Alemany R . Syncytia formation affects the yield and cytotoxicity of an adenovirus expressing a fusogenic glycoprotein at a late stage of replication. Gene Therapy 2008; 15: 1240–1245.
Villanueva A, Garcia C, Paules AB, Vicente M, Megias M, Reyes G et al. Disruption of the antiproliferative TGF-beta signaling pathways in human pancreatic cancer cells. Oncogene 1998; 17: 1969–1978.
Bauerschmitz GJ, Lam JT, Kanerva A, Suzuki K, Nettelbeck DM, Dmitriev I et al. Treatment of ovarian cancer with a tropism modified oncolytic adenovirus. Cancer Res 2002; 62: 1266–1270.
Dmitriev I, Krasnykh V, Miller CR, Wang M, Kashentseva E, Mikheeva G 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.
Olinga P, Merema MT, de Jager MH, Derks F, Melgert BN, Moshage H et al. Rat liver slices as a tool to study LPS-induced inflammatory response in the liver. J Hepatol 2001; 35: 187–194.
Acknowledgements
We thank Blanca Luena and Eduard Serra for their technical assistance. We also thank pathologist Teresa Serrano from the Department of Pathology of the Hospital Universitari de Bellvitge, Spain, for procurement of human liver biopsies. JJ Rojas was supported by a predoctoral fellowship (FI) granted by the Generalitat de Catalunya. This work was supported by BIO2008-04692-C03-01 from the Ministerio de Ciencia y Tecnología of the Government of Spain (RA), EU 6th FP research contract 18700 (Theradpox, RA), 2005 SGR 00727 from the Departament d’Universitats, Recerca i Societat de la Informació of the Generalitat de Catalunya and by Mutua Madrileña Medical Research Foundation. R. Alemany is a member of the Network of Cooperative Research on Cancer (C03-10), Instituto de Salud Carlos III of the Ministerio de Sanidad y Consumo, Government of Spain. A Hemminki is K. Albin Johansson Research Professor of the Foundation for the Finnish Cancer Institute and supported by the European Research Council, HUCH Research Funds (EVO), Sigrid Juselius Foundation, Academy of Finland, Biocentrum Helsinki and Finnish Cancer Society.
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Rojas, J., Cascallo, M., Guedan, S. et al. A modified E2F-1 promoter improves the efficacy to toxicity ratio of oncolytic adenoviruses. Gene Ther 16, 1441–1451 (2009). https://doi.org/10.1038/gt.2009.103
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DOI: https://doi.org/10.1038/gt.2009.103
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