An adenovirus mutant that replicates selectively in p53-deficient human tumor cells

Science

Volumn 274, Issue 5286, 1996, Pages 373-376

  Bischoff, James R ^a,b   Kirn, David H ^a   Williams, Angelica ^a   Heise, Carla ^a   Horn, Sharon ^a   Muna, Mike ^a   Ng, Lelia ^a,b   Nye, Julie A ^a   Sampson Johannes, Adam ^a   Fattaey, Ali ^a   McCormick, Frank ^a  

^a ONYX PHARMACEUTICALS   (United States)

^b SUGEN INC   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PROTEIN P53; VIRUS PROTEIN;

ADENOVIRUS; ANIMAL MODEL; ARTICLE; CLINICAL TRIAL; CONTROLLED CLINICAL TRIAL; CONTROLLED STUDY; CYTOPATHOGENIC EFFECT; DELETION MUTANT; GENE MUTATION; HEAD AND NECK CARCINOMA; HUMAN; HUMAN CELL; MOUSE; NONHUMAN; NUDE MOUSE; PHASE 1 CLINICAL TRIAL; PRIORITY JOURNAL; RAT; SQUAMOUS CELL CARCINOMA; TUMOR CELL CULTURE; TUMOR SUPPRESSOR GENE; UTERINE CERVIX CARCINOMA; VIRUS MUTANT; VIRUS REPLICATION;

ADENOVIRUS E1B PROTEINS; ADENOVIRUSES, HUMAN; ANIMALS; CYTOPATHOGENIC EFFECT, VIRAL; GENE DELETION; GENES, P53; HEAD AND NECK NEOPLASMS; HUMANS; MICE; MICE, NUDE; MUTATION; NEOPLASM TRANSPLANTATION; NEOPLASMS, EXPERIMENTAL; SIGMODONTINAE; TRANSPLANTATION, HETEROLOGOUS; TUMOR CELLS, CULTURED; TUMOR SUPPRESSOR PROTEIN P53; VIRUS REPLICATION;

ADENOVIRIDAE; ANIMALIA; HUMAN ADENOVIRUS; MUS MUSCULUS;

EID: 0344188096     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5286.373     Document Type: Article

References (33)

1. E. White, Nature 371, 21 (1994).
2. S. C. Righetti et al., Cancer Res. 56, 689 (1996); J. S. Kovack et al., Proc. Natl. Acad. Sci. U.S.A 93, 1093 (1996).
3. S. C. Righetti et al., Cancer Res. 56, 689 (1996); J. S. Kovack et al., Proc. Natl. Acad. Sci. U.S.A 93, 1093 (1996).
4. T. A. Van Dyke, Semin. Cancer Biol. 5, 47 (1994); E. Moran, FASEB J. 7, 880 (1993); P. A. Boulanger and G. E. Blair, Biochem. J. 275, 281 (1991); N. Dyson and E. Harlow, Cancer Surv. 12, 161 (1992); E. White and L. R. Gooding, in Apoptosis II: The Molecular Basis of Apoptosis in Disease (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 111-141.
5. T. A. Van Dyke, Semin. Cancer Biol. 5, 47 (1994); E. Moran, FASEB J. 7, 880 (1993); P. A. Boulanger and G. E. Blair, Biochem. J. 275, 281 (1991); N. Dyson and E. Harlow, Cancer Surv. 12, 161 (1992); E. White and L. R. Gooding, in Apoptosis II: The Molecular Basis of Apoptosis in Disease (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 111-141.
6. T. A. Van Dyke, Semin. Cancer Biol. 5, 47 (1994); E. Moran, FASEB J. 7, 880 (1993); P. A. Boulanger and G. E. Blair, Biochem. J. 275, 281 (1991); N. Dyson and E. Harlow, Cancer Surv. 12, 161 (1992); E. White and L. R. Gooding, in Apoptosis II: The Molecular Basis of Apoptosis in Disease (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 111-141.
7. T. A. Van Dyke, Semin. Cancer Biol. 5, 47 (1994); E. Moran, FASEB J. 7, 880 (1993); P. A. Boulanger and G. E. Blair, Biochem. J. 275, 281 (1991); N. Dyson and E. Harlow, Cancer Surv. 12, 161 (1992); E. White and L. R. Gooding, in Apoptosis II: The Molecular Basis of Apoptosis in Disease (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 111-141.
8. T. A. Van Dyke, Semin. Cancer Biol. 5, 47 (1994); E. Moran, FASEB J. 7, 880 (1993); P. A. Boulanger and G. E. Blair, Biochem. J. 275, 281 (1991); N. Dyson and E. Harlow, Cancer Surv. 12, 161 (1992); E. White and L. R. Gooding, in Apoptosis II: The Molecular Basis of Apoptosis in Disease (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 111-141.
9. A. J. van der Eb, C. Mulder, F. L. Graham, A. Houweling, Gene 2, 115 (1977); A. Houweling, P. J. van den Elsen, A. J. van der Eb, Virology 105, 537 (1930); H. Land, L. F. Parada, R. A. Weinberg, Nature 304, 596 (1983); H. E. Ruley, ibid., p. 602.
10. A. J. van der Eb, C. Mulder, F. L. Graham, A. Houweling, Gene 2, 115 (1977); A. Houweling, P. J. van den Elsen, A. J. van der Eb, Virology 105, 537 (1930); H. Land, L. F. Parada, R. A. Weinberg, Nature 304, 596 (1983); H. E. Ruley, ibid., p. 602.
11. A. J. van der Eb, C. Mulder, F. L. Graham, A. Houweling, Gene 2, 115 (1977); A. Houweling, P. J. van den Elsen, A. J. van der Eb, Virology 105, 537 (1930); H. Land, L. F. Parada, R. A. Weinberg, Nature 304, 596 (1983); H. E. Ruley, ibid., p. 602.
12. A. J. van der Eb, C. Mulder, F. L. Graham, A. Houweling, Gene 2, 115 (1977); A. Houweling, P. J. van den Elsen, A. J. van der Eb, Virology 105, 537 (1930); H. Land, L. F. Parada, R. A. Weinberg, Nature 304, 596 (1983); H. E. Ruley, ibid., p. 602.
13. P. R. Yew and A. J. Berk, Nature 357, 82 (1992); C. C. Kao, P. R. Yew, A. J. Berk, Virology 179, 806 (1990).
14. P. R. Yew and A. J. Berk, Nature 357, 82 (1992); C. C. Kao, P. R. Yew, A. J. Berk, Virology 179, 806 (1990).
15. D. D. Barker and A. J. Berk, Virology 156, 107 (1987).
16. M. Scheffner, K. Munger, J. C. Byrne, P. M. Howley, Proc. Natl. Acacl. Sci. U.S.A. 88, 5523 (1991); T. Crook, D. Wrede, K. H. Vousden, Oncogene 6, 873 (1991).
17. M. Scheffner, K. Munger, J. C. Byrne, P. M. Howley, Proc. Natl. Acacl. Sci. U.S.A. 88, 5523 (1991); T. Crook, D. Wrede, K. H. Vousden, Oncogene 6, 873 (1991).
18. L. Diller et al., Mol. Cell. Biol. 10, 5772 (1990).
19. J. R. Bischoff et al., unpublished observations.
20. D. Kirn et al., unpublished observations.
21. E. G. Van Meir et al., Cancer Res. 54, 649 (1994).
22. B. Ruggeri et al., Oncogene 7, 1503 (1992).
23. 2-terminus of E1B 55K, and the construct was subcloned into pCMVneo. This clone was designated pCMVFLSS. Another construct was made in which amino acids 216 through 354 of the Myctagged full-length E1B 55K were deleted. This clone, which was expressed in U4.5H cells, was designated pCMV55D4.5. The inserts of pCMVFLSSk and pCMV55D4.5 were sequenced to confirm that they were correct (9).
24. W. J. Slichenmeyer, W. G. Nelson, R. J. Slebos, M. B. Kastan, Cancer Res. 53, 4164 (1993).
25. T. Dobner, N. Horikoshi, S. Rubenwolf, T. Shenk, Science 272, 1470 (1996).
26. 9 PFU of dl1520 per animal. No evidence of toxicity was seen in these animals, as assayed by animal weight gain, serum chemistries, and organ histopathology (including the liver, lung, kidneys, heart, gastrointestinal tract, and brain).
27. Phase I tests of dl1520 (ONYX-015) began in April 1996 at the University of Texas, San Antonio (by Daniel Von Hoff) and the Beatson Institute, Glasgow, Scotland (by Stanley Kaye).
28. Plates were scraped into 1 ml of media and frozen. Lysates were prepared by three cycles of freezing and thawing, followed by a 30-s pulse in a sonicator water bath. Serial dilutions of the lysates were titered on HEK293 cells (human embryonic kidney cells expressing the E1 region of Ad2).
29. 35S]methionine for 2 hours at 37°C. The cells were lysed in 50 mM Hepes (pH 7.9), 250 mM NaCl, 50 mM NaF, 5 mM EDTA, 0.1 mM Na orthovanadate, and 0.1% Triton X-100. The protein concentration was determined by Bradford analysis. Equal amounts of protein were immunoprecipitated with the appropriate antibody for 1 hour at 4°C, protein G-Sepharose (Sigma) was added, and the samples were incubated (or an additional 30 to 60 min at 4°C. The immunocomplexes were washed three times in lysis buffer, resuspended in SDS sample buffer, and resolved on 10% polyacrylamide gels. The gels were fixed, dried, and subjected to autoradiography.
30. Lysates were prepared 36 hours after transfection and chloramphenicol acetyl transferase (CAT] (Boehringer-Mannheim), and luciferase (Promega) assays were performed according to the manufacturer's instructions.
31. 9 PFU of dl1520 (n = 10 UB7 tumors and 5 C33A tumors), Ad2 (n = 10 tumors), or UV-inactivated Ad2 (n = 6 tumors) divided equally into four tumor quadrants (15 μl per quadrant) every other day for three total doses. Tumor volumes were recorded weekly until termination of the study. At the time of termination, the tumor volume ratio was calculated as follows: (tumor volume at study termination)/(tumor volume at the time of virus injection). The unpaired t test (two-tailed) was used to compare final tumor volume ratios in various groups.
32. Immunohistochemistry was performed on formalin-fixed paraffin-embedded tumors that had been cut into 4-μm sections, hydrated, and digested with pronase. The primary antibody (MAB805, Chemicon International) is specific for all 41 serotypes of adenovirus hexon protein. Tissues were incubated for 1 hour at 35°C with an antibody dilution of 1:1000. A biotinylated goat secondary antibody to mouse immunoglobulin was then applied, followed by a streptavidin-horseradish peroxidase conjugate. Diaminobenzidine was used as the chromagen, and slides were counterstained with hematoxylin.
33. We thank A. Berk (University of California, Los Angeles) for providing wild-type adenovirus and dl1520; E. White (Rutgers University) for a cDNA encoding full-length E1B 55K polypeptide; M. Kastan (Johns Hopkins University) for RKO cells; J, Hassell (Mc-Master University) for HEK293 cells; D. Von Moff, G. Mangold, and D. Dexter (San Antonio Cancer Treatment and Research Center) for help with animal models; J. Olesch for technical help; and C. Maack for many stimulating discussions.