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Adenovirus-mediated tissue-targeted expression of the HSVtk gene for the treatment of breast cancer

Gene Therapy volume 6, pages 854–864 (1999)Cite this article

Abstract

In an effort to develop a genetic therapy for the treatment of breast cancer, we constructed adenoviral vectors containing either the beta-galactosidase (β-gal) reporter gene or the herpes simplex thymidine kinase (HSVtk) suicide gene driven by breast tissue-specific promoters. We utilized upstream regulatory sequences from either the human alpha-lactalbumin (hALA) gene, or the ovine beta-lactoglobulin (oBLG) gene in these vector constructs to target expression of heterologous genes transcriptionally to breast cancer cells both in vitro and in vivo. Data derived from breast tissue-specific reporter vectors in vitro demonstrate that expression from the hALA and oBLG promoters are indeed specific for breast cells (T47D, MCF-7, ZR75–1) when compared with non-breast cells (U2OS, HeLa). Moreover, these vectors displayed tumor cell specificity when compared with the normal MCF-10A breast cell line. These vectors also displayed breast tissue specificity when injected systemically (i.v.) into lactating Balb/c mice, which suggests that these promoters maintain their tissue- specific expression pattern within the context of the adenoviral genome in vivo. Tumors, derived from T47D human breast cancer cells, were established in nude mice and injected with either the tissue-specific reporter or suicide vectors. Results from tumors injected (i.t.) with reporter adenoviruses demonstrate that these promoters are active in T47D cells when grown as established tumors and we observed a marked regression of tumors injected with suicide vectors and treated systemically with gancyclovir (150 mg/kg/day) when compared with control animals. Moreover, mouse survival was prolonged after 35 days in mice undergoing therapy with the suicide vectors in conjunction with gancyclovir when compared with the control animals. These data suggest that the transcriptionally targeted hALA or oBLG driven expression of the HSVtk gene may be a feasible therapy for the treatment of human breast cancer.

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References

  1. Weichselbaum R, Kufe D . Gene therapy of cancer Lancet 1997; 349: SII10–SII102

  2. Miller N, Whelan J . Progress in transcriptionally targeted and regulatable vectors for genetic therapy Hum Gene Ther 1997 8: 803–815

    Article  CAS  Google Scholar 

  3. Shirakawa T, Sung-Chu K, Gardner TA, Jun C, Tadayuki M, Gotoh A, Chung LWK, Kao C . In vivo suppression of osteosarcoma pulmonary metastasis with intravenous osteocalcin promoter-based toxic gene therapy Cancer Gene Ther 1998 5: 274–280

    CAS  PubMed  Google Scholar 

  4. Cohen C, Sharkey FE, Shulman G, Uthman E, Budgeon L . Tumor-associated antigens in breast carcinomas Cancer 1987 60: 1294–1298

    Article  CAS  Google Scholar 

  5. De Almeida PC, Pestana CB . Immunohistochemical markers in the identification of metastic breast cancer Breast Cancer Res Treat 1992 21: 201–210

    Article  CAS  Google Scholar 

  6. Thean ET, Toh B . Serum human alpha-lactalbumin as a marker for breast cancer Br J Cancer 1990 61: 773–775

    Article  CAS  Google Scholar 

  7. Lee AK, Delellis RA, Rosen PP, Herbert-Stanton T, Tallberg K, Garcia C, Wolfe H . Alpha-lactalbumin as an immunohistochemical marker for metastatic breast carcinomas Am J Surg Pathol 1984 8: 93–100

    Article  CAS  Google Scholar 

  8. Collette J, Hendrick JC, Jaspar J, Franchimont P . Presence of alpha-lactalbumin, epidermal growth factor, epithelial membrane antigen, and gross cystic disease fluid protein (15000 daltons) in breast cyst fluid Cancer Res 1986 46: 3728–3733

    CAS  PubMed  Google Scholar 

  9. Simickova M, Lang BA, Rejthar A, Cernoch M, Sakalovk J, Pacovsky Z . Immunohistochemical localization of alpha-lactalbumin in human breast cancer tissue Neoplasma 1989 36: 11–20

    CAS  PubMed  Google Scholar 

  10. Walker RA . The demonstration of alpha-lactalbumin in human breast carcinomas J Pathol 1979 129: 37–42

    Article  CAS  Google Scholar 

  11. Eden JA, Wren BG . Hormone replacement therapy: a review Cancer Treat Rev 1996 22: 335–343

    Article  CAS  Google Scholar 

  12. Findlay J, Brew K . The complete amino acid sequence of human alpha-lactalbumin Eur J Biochem 1972 27: 65–86

    Article  CAS  Google Scholar 

  13. Quirk SJ, Gannell JE, Funder JW . Alpha-lactalbumin production by rat mammary gland: prepartum emergence of resistance to glucocorticoids and progestins Mol Cell Biol 1988 58: 183–189

    CAS  Google Scholar 

  14. Maschio A et al. Transgenic mice carrying the guinea-pig alpha-lactalbumin gene transcribe milk protein genes in their sebaceous glands during lactation Biochem J 1991 275: 459–467

    Article  CAS  Google Scholar 

  15. Stacy A et al. Use of double-replacment gene targeting to replace the murine ALA gene with its human counterpart in embryonic stem cells and mice. Mol Cell Biol 1994; 14: 1009–1016

  16. Stacy A et al. Lactation is disrupted by alpha-lactalbumin deficiency and can be restored by human alpha-lactalbumin gene replacement in mice. Proc Natl Acad Sci USA 1995; 92: 2835–2839

  17. Moolten FL, Wells M . Curability of tumors bearing herpes thymidine kinase genes transferred by retroviral vectors J Natl Cancer Inst 1990 82: 297–300

    Article  CAS  Google Scholar 

  18. Ali S, Clark J . Characterization of the gene encoding ovine beta-lactoglobulin: similarity to the genes for retinol binding protein and secretory proteins J Mol Biol 1988 199: 415–426

    Article  CAS  Google Scholar 

  19. Ali S, McClenaghan M, Simons PJ, Clark JA . Characterization of the alleles encoding ovine beta-lactoglobulins A and B J Mol Biol 1990 91: 201–207

    CAS  Google Scholar 

  20. Mink S et al. A mammary cell-specific enhancer in mouse mammary tumor virus DNA is composed of multiple regulatory elements including binding sites for CTF/NFI and a novel transcription factor, mammary cell-activating factor Mol Cell Biol 1992 12: 4906–4918

    Article  CAS  Google Scholar 

  21. Bruce C et al. Position-independent expression of the ovine beta-lactoglobulin gene in transgenic mice Biochem J 1992 286: 31–39

    Article  Google Scholar 

  22. Mercier JC, Vilotte JL . Structure and function of milk protein genes J Dairy Sci 1993 76: 3079–3098

    Article  CAS  Google Scholar 

  23. Harris S et al. Developmental regulation of the sheep beta-lactoglobulin gene in the mammary gland of transgenic mice Dev Genet 1991 12: 299–307

    Article  CAS  Google Scholar 

  24. Curiel DT, Pilewski JM, Albelda SM . Gene therapy approaches for inherited and acquired lung diseases Am J Respir Cell Mol Biol 1996 14: 1–18

    Article  CAS  Google Scholar 

  25. Esandi MC et al. Gene therapy of experimental malignant mesothelioma using adenovirus vectors encoding the HSVtk gene Gene Therapy 1997 4: 280–287

    Article  CAS  Google Scholar 

  26. Connelly S et al. Complete short-term correction of canine hemophilia A by in vivo gene therapy Blood 1996 88: 3846–3853

    CAS  PubMed  Google Scholar 

  27. Chen SH et al. Combination suicide and cytokine gene therapy for hepatic metastases of colon carcinoma: sustained antitumor immunity prolongs animal survival Cancer Res 1996 56: 3758–3762

    CAS  PubMed  Google Scholar 

  28. Zhu N, Liggitt D, Liu Y, Debs R . Systemic gene expression after intravenous DNA delivery into adult mice Science 1993 261: 209–211

    Article  CAS  Google Scholar 

  29. Kass-Eisler A et al. The impact of developmental stage, route of injection and the immune system on adenovirus-mediated gene transfer Gene Therapy 1994 1: 395–402

    CAS  PubMed  Google Scholar 

  30. Smiley RS et al. Establishment of parameters for optimal transduction efficiency and antitumor effects with purified high-titer HSVtk retroviral vector in established solid tumors Hum Gene Ther 1997 8: 965–977

    Article  CAS  Google Scholar 

  31. Stow ND . Cloning of a DNA fragment from the left-hand terminus of the adenovirus type 2 genome and its use in site-directed mutagenesis J Virol 1981 37: 171–180

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Swaminathan S, Thimmapaya B . Transactivation of adenovirus E2-early promoter by E1a and E4 6/7 in the context of the viral chromosome J Mol Biol 1996 258: 736–746

    Article  CAS  Google Scholar 

  33. Graham FL, Prevec L . Manipulation of adenovirus vectors. In: Murray EJ (ed). Gene Transfer and Expression Protocols Humana Press: New York 1991 pp109–128

    Book  Google Scholar 

  34. Bradford MM . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 1976 72: 248–254

    Article  CAS  Google Scholar 

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Author notes

  1. B Thimmapaya and S A Weitzman: B Thimmapaya and SA Weitzman are senior co-authors

Authors and Affiliations

  1. Robert H Lurie Cancer Center, Northwestern University Medical School, Chicago, IL, USA

    L M Anderson, S Swaminathan, I Zackon, A-K Tajuddin, B Thimmapaya & S A Weitzman

  2. Department of Microbiology and Immunology, Northwestern University Medical School, Chicago, IL, USA

    L M Anderson, S Swaminathan & B Thimmapaya

  3. Division of Hematology and Oncology, Department of Medicine, Northwestern University Medical School, Chicago, IL, USA

    I Zackon, A-K Tajuddin & S A Weitzman

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  1. L M Anderson
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  2. S Swaminathan
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  5. B Thimmapaya
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Anderson, L., Swaminathan, S., Zackon, I. et al. Adenovirus-mediated tissue-targeted expression of the HSVtk gene for the treatment of breast cancer. Gene Ther 6, 854–864 (1999). https://doi.org/10.1038/sj.gt.3300909

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