Loss of p14ARF in tumor cells facilitates replication of the adenovirus mutant dl1520 (ONYX-015)

Abstract

The adenovirus mutant dl1520 (ONYX-015) does not express the E1B-55K protein that binds and inactivates p53. This virus replicates in tumor cells with mutant p53, but not in normal cells with functional p53. Although intra-tumoral injection of dl1520 shows promising responses in patients with solid tumors, previous in vitro studies have not established a close correlation between p53 status and dl1520 replication. Here we identify loss of p14ARF as a mechanism that allows dl1520 replication in tumor cells retaining wild-type p53. We demonstrate that the re-introduction of p14ARF into tumor cells with wild-type p53 suppresses replication of dl1520 in a p53-dependent manner. Our study supports the therapeutic use of dl1520 in tumors with lesions within the p53 pathway other than mutation of p53.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Response of p53, p21WAF1/CIP1 and Mdm2 in HCT 116 TP53+/+ cells following adenovirus infection.
Figure 2: The adenovirus E1A protein activates p53 independently of p14ARF in HCT 116 TP53+/+ cells.
Figure 3: p14ARF protein levels in normal cells and A2780 cells.
Figure 4: Effect of p14ARF re-introduction into HCT116 TP53+/+ cells on p53.
Figure 5: p14ARF protects HCT116 TP53+/+ cells from lysis by dl1520 replication.

References

  1. 1

    Fearon, E.R., Hamilton, S.R. & Vogelstein, B. Clonal analysis of human colorectal tumors. Science 238, 193–197 (1987).

  2. 2

    Chang, F., Syrjanen, S. & Syrjanen, K. Implications of the p53 tumor-suppressor gene in clinical oncology. J. Clin. Oncol. 13, 1009–1022 (1995).

  3. 3

    Lowe, S.W. et al. p53 status and the efficacy of cancer therapy in vivo. Science 266, 807–810 (1994).

  4. 4

    Bergh, J., Norberg, T., Sjogren, S., Lindgren, A. & Holmberg, L. Complete sequencing of the p53 gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy. Nature Med. 1, 1029–1034 (1995).

  5. 5

    Barker, D.D. & Berk, A.J. Adenovirus proteins from both E1B reading frames are required for transformation of rodent cells by viral infection and DNA transfection. Virology 156, 107–121 (1987).

  6. 6

    Ganly, I. et al. A phase I study of Onyx-015, an E1B attenuated adenovirus, administered intratumorally to patients with recurrent head and neck cancer. Clin. Cancer Res. 6, 798–806 (2000).

  7. 7

    Khuri, F.R. et al. A controlled trial of intratumoral ONYX-015, a selectively-replicating adenovirus, in combination with cisplatin and 5-fluorouracil in patients with recurrent head and neck cancer. Nature Med. 6, 879–885 (2000).

  8. 8

    Heise, C. et al. ONYX-015, an E1B gene-attenuated adenovirus, causes tumor-specific cytolysis and antitumoral efficacy that can be augmented by standard chemotherapeutic agents. Nature Med. 3, 639–645 (1997).

  9. 9

    Bischoff, J.R. et al. An adenovirus mutant that replicates selectively in p53-deficient human tumor cells. Science 274, 373–376 (1996).

  10. 10

    Goodrum, F.D. & Ornelles, D.A. p53 status does not determine outcome of E1B 55-kilodalton mutant adenovirus lytic infection. J. Virol. 72, 9479–9490 (1998).

  11. 11

    Turnell, A.S., Grand, R.J. & Gallimore, P.H. The replicative capacities of large E1B-null group A and group C adenoviruses are independent of host cell p53 status. J. Virol. 73, 2074–2083 (1999).

  12. 12

    Rothmann, T., Hengstermann, A., Whitaker, N.J., Scheffner, M. & zur Hausen, H. Replication of ONYX-015, a potential anticancer adenovirus, is independent of p53 status in tumor cells. J. Virol. 72, 9470–9478 (1998).

  13. 13

    Harada, J.N. & Berk, A.J. p53-independent and -dependent requirements for E1B-55K in adenovirus type 5 replication. J. Virol. 73, 5333–5344 (1999).

  14. 14

    Piette, J., Neel, H. & Marechal, V. Mdm2: keeping p53 under control. Oncogene 15, 1001–1010 (1997).

  15. 15

    Kamijo, T. et al. Functional and physical interactions of the ARF tumor suppressor with p53 and Mdm2. Proc. Natl Acad. Sci. USA 95, 8292–8297 (1998).

  16. 16

    Pomerantz, J. et al. The Ink4a tumor suppressor gene product, p19Arf, interacts with MDM2 and neutralizes MDM2's inhibition of p53. Cell 92, 713–723 (1998).

  17. 17

    Waldman, T., Kinzler, K.W. & Vogelstein, B. p21 is necessary for the p53-mediated G1 arrest in human cancer cells. Cancer Res. 55, 5187–5190 (1995).

  18. 18

    Bunz, F. et al. Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 282, 1497–1501 (1998).

  19. 19

    Di Leonardo, A., Linke, S.P., Clarkin, K. & Wahl, G.M. DNA damage triggers a prolonged p53-dependent G1 arrest and long-term induction of Cip1 in normal human fibroblasts. Genes Dev. 8, 2540–2551 (1994).

  20. 20

    Barak, Y., Juven, T., Haffner, R. & Oren, M. mdm2 expression is induced by wild type p53 activity. EMBO J. 12, 461–468 (1993).

  21. 21

    Stott, F.J. et al. The alternative product from the human CDKN2A locus, p14(ARF), participates in a regulatory feedback loop with p53 and MDM2. EMBO J. 17, 5001–5014 (1998).

  22. 22

    Bates, S. et al. p14ARF links the tumour suppressors RB and p53. Nature 395, 124–125 (1998).

  23. 23

    de Stanchina, E. et al. E1A signaling to p53 involves the p19(ARF) tumor suppressor. Genes Dev. 12, 2434–2442 (1998).

  24. 24

    Weber, J.D., Taylor, L.J., Roussel, M.F., Sherr, C.J. & Bar-Sagi, D. Nucleolar Arf sequesters Mdm2 and activates p53. Nature Cell Biol. 1, 20–26 (1999).

  25. 25

    Zhang, Y. & Xiong, Y. Mutations in human ARF exon 2 disrupt its nucleolar localization and impair its ability to block nuclear export of MDM2 and p53. Mol. Cell 3, 579–591 (1999).

  26. 26

    Brown, R. et al. Increased accumulation of p53 protein in cisplatin-resistant ovarian cell lines. Int. J. Cancer 55, 678–684 (1993).

  27. 27

    Steegenga, W.T., Riteco, N. & Bos, J.L. Infectivity and expression of the early adenovirus proteins are important regulators of wild-type and DeltaE1B adenovirus replication in human cells. Oncogene 18, 5032–5043 (1999).

  28. 28

    Billon, N., van Grunsven, L.A. & Rudkin, B.B. The CDK inhibitor p21WAF1/Cip1 is induced through a p300-dependent mechanism during NGF-mediated neuronal differentiation of PC12 cells. Oncogene 13, 2047–2054 (1996).

  29. 29

    Roth, J., Dobbelstein, M., Freedman, D.A., Shenk, T. & Levine, A.J. Nucleo-cytoplasmic shuttling of the hdm2 oncoprotein regulates the levels of the p53 protein via a pathway used by the human immunodeficiency virus rev protein. EMBO J. 17, 554–564 (1998).

  30. 30

    Dobner, T., Horikoshi, N., Rubenwolf, S. & Shenk, T. Blockage by adenovirus E4orf6 of transcriptional activation by the p53 tumor suppressor. Science 272, 1470–1473 (1996).

  31. 31

    Tao, W. & Levine, A.J. P19(ARF) stabilizes p53 by blocking nucleo-cytoplasmic shuttling of Mdm2. Proc. Natl Acad. Sci. USA 96, 6937–6941 (1999).

  32. 32

    Schmitt, C.A., McCurrach, M.E., de Stanchina, E., Wallace-Brodeur, R.R. & Lowe, S.W. INK4a/ARF mutations accelerate lymphomagenesis and promote chemoresistance by disabling p53. Genes Dev. 13, 2670–2677 (1999).

  33. 33

    Hall, A.R., Dix, B.R., O'Carroll, S.J. & Braithwaite, A.W. p53-dependent cell death/apoptosis is required for a productive adenovirus infection. Nature Med. 4, 1068–1072 (1998).

  34. 34

    Babiss, L.E., Ginsberg, H.S. & Darnell, J.E., Jr. Adenovirus E1B proteins are required for accumulation of late viral mRNA and for effects on cellular mRNA translation and transport. Mol. Cell. Biol. 5, 2552–2558 (1985).

Download references

Acknowledgements

We thank B. Vogelstein and F. Bunz for the HCT116 cell lines; E. Lees for the p14ARF antibody and the pCMV-p14ARFexpression plasmid; H. Jiang, Y.J. Shen and O. Tetsu for technical advice; J. Giovanola and K. Halfmeyer for technical assistance; D. Moore for helping with the statistical analysis; A. Fattaey, L. Johnson, C. O'Shea and J. Lyons for suggestions and reagents; and B. Bastian for discussions. S.J.R. is a recipient of a fellowship as part of the “Gemeinsames Hochschulsonderprogramm III von Bund und Ländern” granted by the German Academic Exchange Service, DAAD. C.H.B. is the recipient of a fellowship granted by the Deutsche Forschungsgemeinschaft (DFG).

Author information

Correspondence to W. Michael Korn.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ries, S., Brandts, C., Chung, A. et al. Loss of p14ARF in tumor cells facilitates replication of the adenovirus mutant dl1520 (ONYX-015). Nat Med 6, 1128–1133 (2000). https://doi.org/10.1038/80466

Download citation

Further reading