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Intraperitoneal gene therapy by rAAV provides long-term survival against epithelial ovarian cancer independently of survivin pathway

Gene Therapy volume 14, pages 138–146 (2007)Cite this article

Abstract

Epithelial ovarian carcinoma is the leading cause of death from gynecological malignancies. Owing to the lack of an effective screening method, insidious onset, and non-specific symptoms, a majority of women present with advanced stage disease. Despite improvements from cytoreductive surgery and chemotherapy, recurrent disease remains a formidable challenge. In the present study, we demonstrate for the first time that stable intra-abdominal genetic transfer of endostatin and angiostatin (E+A) by recombinant adeno-associated virus (rAAV) provides sustained antitumor effects on the growth and dissemination of epithelial ovarian cancer in a mouse model. Further, when combined with paclitaxel (taxol), the effect of this therapy was dramatically increased and resulted in long-term tumor-free survival overcoming prior limitations of chemotherapy and gene therapy. The combined effects of angiosuppressive therapy and chemotherapy were found to be independently of survivin pathway. Evidence for the superior effects of the combination therapy was indicated by significantly lower ascites volume with less hemorrhage and tumor conglomerates, lower ascites vascular endothelial growth factor, higher tumor cell apoptosis and decreased blood vasculature, and long-term disease-free survival. Histopathology of visceral organs and liver enzyme assays indicated no toxicity or pathology.

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References

  1. Jemal A, Siegel R, Ward E, Murray T, Xu J, Smigal C . Cancer statistics, 2006. CA Cancer J Clin 2006; 56: 106–130.

    Article  Google Scholar 

  2. Cannistra SA . Cancer of the ovary. N Engl J Med 2004; 351: 2519–2529.

    Article  CAS  Google Scholar 

  3. Berkenblit A, Cannistra SA . Advances in the management of epithelial ovarian cancer. J Reprod Med 2005; 50: 426–438.

    PubMed  Google Scholar 

  4. Holschneider CH, Berek JS . Ovarian cancer: epidemiology, biology, and prognostic factors. Semin Surg Oncol 2000; 19: 3–10.

    Article  CAS  Google Scholar 

  5. Young RC, Pecorelli S . Management of early ovarian cancer. Semin Oncol 1998; 25: 335–339.

    CAS  PubMed  Google Scholar 

  6. Ozols RF . Future directions in the treatment of ovarian cancer. Semin Oncol 2002; 29: 32–42.

    Article  CAS  Google Scholar 

  7. Folkman J . Tumor angiogenesis: therapeutic implications. N Engl J Med 1971; 285: 1182–1186.

    Article  CAS  Google Scholar 

  8. Folkman J . Anti-angiogenic gene therapy. Proc Natl Acad Sci USA 1998; 95: 9064–9066.

    Article  CAS  Google Scholar 

  9. Li L, Wang L, Zhang W, Tang B, Zhang J, Song H et al. Correlation of serum VEGF levels with clinical stage, therapy efficacy, tumor metastasis and patient survival in ovarian cancer. Anticancer Res 2004; 24: 1973–1979.

    CAS  PubMed  Google Scholar 

  10. Raspollini MR, Amunni G, Villanucci A, Baroni G, Boddi V, Taddei GL . Prognostic significance of microvessel density and vascular endothelial growth factor expression in advanced ovarian serous carcinoma. Int J Gynecol Cancer 2004; 14: 815–823.

    Article  CAS  Google Scholar 

  11. Ponnazhagan S, Curiel DT, Shaw DR, Alvarez R, Siegal GP . Adeno-associated viral vectors for cancer gene therapy. Cancer Res 2001; 61: 6313–6321.

    CAS  PubMed  Google Scholar 

  12. Boocock CA, Charnock-Jones DS, Sharkey AM, McLaren J, Barker PJ, Wright KA et al. Expression of vascular endothelial growth factor and its receptors flt and KDR in ovarian carcinoma. J Natl Cancer Inst (Bethesda) 1995; 87: 506–516.

    Article  CAS  Google Scholar 

  13. Olson TA, Mohanraj D, Carson LF, Ramakrishnan S . Vascular permeability factor gene expression in normal and neoplastic human ovaries. Cancer Res 1994; 54: 276–280.

    CAS  PubMed  Google Scholar 

  14. Schondorf T, Kurbacher CM, Gohring UJ, Benz C, Becker M, Sartorius J et al. Induction of MDR1-gene expression by antineoplastic agents in ovarian cancer cell lines. Anticancer Res 2002; 22: 2199–2203.

    PubMed  Google Scholar 

  15. Mozzetti S, Ferlini C, Concolino P, Filippetti F, Raspaglio G, Prislei S et al. Class III beta-tubulin overexpression is a prominent mechanism of paclitaxel resistance in ovarian cancer patients. Clin Cancer Res 2005; 11: 298–305.

    CAS  PubMed  Google Scholar 

  16. Nyberg P, Heikkila P, Sorsa T, Luostarinen J, Heljasvaara R, Stenman UH et al. Endostatin inhibits human tongue carcinoma cell invasion and intravasation and blocks the activation of matrix metalloprotease-2, 9 and -13. J Biol Chem 2003; 278: 22404–22411.

    Article  CAS  Google Scholar 

  17. O'Reilly MS, Wiederschain D, Stetler-Stevenson WG, Folkman J, Moses MA . Regulation of angiostatin production by matrix metalloproteinase-2 in a model of concomitant resistance. J Biol Chem 1999; 274: 29568–29571.

    Article  CAS  Google Scholar 

  18. Wickstrom SA, Alitalo K, Keski-Oja J . Endostatin associates with integrin alpha5beta1 and caveolin-1, and activates Src via a tyrosyl phosphatase-dependent pathway in human endothelial cells. Cancer Res 2002; 62: 5580–5589.

    CAS  Google Scholar 

  19. Conlon TJ, Flotte TR . Recombinant adeno-associated virus vectors for gene therapy. Expert Opin Biol Ther 2004; 4: 1093–1101.

    Article  CAS  Google Scholar 

  20. Ponnazhagan S, Mahendra G, Kumar S, Shaw D, Meleth S, Stockardt R et al. Adeno-associated virus-2-mediated anti-angiogenic gene therapy: long-term efficacy of a vector encoding angiostatin and endostatin over vectors encoding a single factor. Cancer Res 2004; 64: 1781–1787.

    Article  CAS  Google Scholar 

  21. Ponnazhagan S, Mahendra G, Lima J, Aldrich W, Jenkins C, Ren C et al. Augmentation of anti-tumor activity of a recombinant adeno-associated virus carcinoembryonic antigen vaccine with plasmid adjuvants. Hum Gene Ther 2004; 15: 856–864.

    Article  CAS  Google Scholar 

  22. Mahendra G, Mahasreshti P, Curiel DT, Stockardt R, Grizzle WE, Alapati V et al. Anti-angiogenic gene therapy through adeno-associated virus 2-mediated stable expression of soluble Flt-1 receptor. Cancer Gene Ther 2005; 12: 26–34.

    Article  CAS  Google Scholar 

  23. Isayeva T, Ren C, Ponnazhagan S . Recombinant adeno-associated virus 2-mediated anti-angiogenic prevention in a mouse model of intraperitoneal ovarian cancer. Clin Cancer Res 2005; 11: 1342–1347.

    CAS  PubMed  Google Scholar 

  24. Lalani AS, Chang B, Lin J, Case SS, Luan B, Wu-Prior WW et al. Anti-tumor efficacy of human angiostatin using liver-mediated adeno-associated virus gene therapy. Mol Ther 2004; 9: 56–66.

    Article  CAS  Google Scholar 

  25. Subramanian IV, Ghebre R, Ramakrishnan S . Adeno-associated virus-mediated delivery of a mutant endostatin suppresses ovarian carcinoma growth in mice. Gene Therapy 2005; 12: 30–38.

    Article  CAS  Google Scholar 

  26. Ling X, Bernacki RJ, Brattain MG, Li F . Induction of survivin expression by taxol (paclitaxel) is an early event, which is independent of taxol-mediated G2/M arrest. J Biol Chem 2004; 279: 15196–15203.

    Article  CAS  Google Scholar 

  27. O'Connor DS, Wall NR, Porter AC, Altieri DC . A p34(cdc2) survival checkpoint in cancer. Cancer Cell 2002; 2: 43–54.

    Article  CAS  Google Scholar 

  28. Kirby TO, Curiel DT, Alvarez RD . Gene therapy for ovarian cancer: progress and potential. Hematol Oncol Clin North Am 2003; 17: 1021–1050.

    Article  Google Scholar 

  29. Nagy JA, Masse EM, Herzberg KT, Meyers MS, Yeo KT, Yeo TK et al. Pathogenesis of ascites tumor growth: vascular permeability factor, vascular hyperpermeability, and ascites fluid accumulation. Cancer Res 1995; 55: 360–368.

    CAS  PubMed  Google Scholar 

  30. Yoneda J, Kuniyasu H, Crispens MA, Price JE, Bucana CD, Fidler IJ . Expression of angiogenesis-related genes and progression of human ovarian carcinomas in nude mice. J Natl Cancer Inst (Bethesda) 1998; 90: 447–454.

    Article  CAS  Google Scholar 

  31. Mesiano S, Ferrara N, Jaffe RB . Role of vascular endothelial growth factor in ovarian cancer: inhibition of ascites formation by immunoneutralization. Am J Pathol 1998; 153: 1249–1256.

    Article  CAS  Google Scholar 

  32. Nakanishi Y, Kodama J, Yoshinouchi M, Tokumo K, Kamimura S, Okuda H et al. The expression of vascular endothelial growth factor and transforming growth factor-β associates with angiogenesis in epithelial ovarian cancer. Int J Gynecol Pathol 1997; 16: 256–262.

    Article  CAS  Google Scholar 

  33. Fujimoto J, Sakaguchi H, Hirose R, Ichigo S, Tamaya T . Biologic implications of the expression of vascular endothelial growth factor subtypes in ovarian carcinoma. Cancer 1998; 83: 2528–2533.

    Article  CAS  Google Scholar 

  34. Sykes JA . Pharmacologically active substances in malignant ascites fluid. Br J Pharmacol 1970; 40: 595.

    Google Scholar 

  35. Greenbaum LM, Grebow P, Johnston M, Prakash A, Semente G . Pepstatin, an inhibitor of leukokinin formation and ascitic fluid accumulation. Cancer Res 1975; 35: 706–710.

    CAS  PubMed  Google Scholar 

  36. Ettinghausen SE, Puri RK, Rosenberg SA . Increased vascular permeability in organs mediated by the systemic administration of lymphokine-activated killer cells and recombinant interleukin-2 in mice. J Natl Cancer Inst (Bethesda) 1998; 80: 177–188.

    Article  Google Scholar 

  37. Ohmura E, Tsushima T, Kamiya Y, Okada M, Onoda N, Shizume K et al. Epidermal growth factor and transforming growth factor α-induce ascitic fluid in mice. Cancer Res 1990; 50: 4915–4917.

    CAS  PubMed  Google Scholar 

  38. Kumar N . Taxol-induced polymerization of purified tubulin. Mechanism of action. J Biol Chem 1981; 256: 10435–10441.

    CAS  PubMed  Google Scholar 

  39. Schiff PB, Fant J, Horwitz SB . Promotion of microtubule assembly in vitro by taxol. Nature 1979; 277: 665–667.

    Article  CAS  Google Scholar 

  40. Schiff PB, Horwitz SB . Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci USA 1980; 77: 1561–1565.

    Article  CAS  Google Scholar 

  41. Schibler MJ, Cabral F . Taxol-dependent mutants of Chinese hamster ovary cells with alterations in alpha- and beta-tubulin. J Cell Biol 1986; 102: 1522–1531.

    Article  CAS  Google Scholar 

  42. Giannakakou P, Sackett DL, Kang YK, Zhan Z, Buters JT, Fojo T et al. Paclitaxel-resistant human ovarian cancer cells have mutant beta-tubulins that exhibit impaired paclitaxel-driven polymerization. J Biol Chem 1997; 272: 17118–17125.

    Article  CAS  Google Scholar 

  43. Snyder RO . Adeno-associated virus-mediated gene delivery. J Gene Med 1999; 1: 166–175.

    Article  CAS  Google Scholar 

  44. Gao GP, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson JM . Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc Natl Acad Sci USA 2002; 99: 11854–11859.

    Article  CAS  Google Scholar 

  45. Gao GP, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson J . Adeno-associated viruses undergo substantial evolution in primates during natural infections. Proc Natl Acad Sci USA 2003; 100: 6081–6086.

    Article  CAS  Google Scholar 

  46. Thomas JP, Arzoomanian RZ, Alberti D, Marnocha R, Lee F, Friedl A et al. Phase I pharmacokinetic and pharmacodynamic study of recombinant human endostatin in patients with advanced solid tumors. J Clin Oncol 2003; 21: 223–231.

    Article  CAS  Google Scholar 

  47. Ye X, Rivera VM, Zoltick P, Cerasoli Jr F, Schnell MA, Gao G et al. Regulated delivery of therapeutic proteins after in vivo somatic cell gene transfer. Science 1999; 283: 88–91.

    Article  CAS  Google Scholar 

  48. Rivera VM, Ye X, Courage NL, Sachar J, Cerasoli Jr F, Wilson JM et al. Long-term regulated expression of growth hormone in mice after intramuscular gene transfer. Proc Natl Acad Sci USA 1999; 15: 8657–8662.

    Article  Google Scholar 

  49. Tinwell H, Ashby J . Genetic toxicity and potential carcinogenicity of taxol. Carcinogenesis 1994; 8: 1499–1501.

    Article  Google Scholar 

  50. Ozkan L, Egeli U, Tunca B, Aydemir N, Cecener G, Akpinar G et al. Investigation of genotoxic effects of taxol plus radiation on mice bone marrow cells. Teratogen Carcinogen Mutagen 2002; 22: 1–11.

    Article  CAS  Google Scholar 

  51. Zolotukhin S, Byrne B, Mason E, Zolotukhin I, Potter M, Chesnut K et al. Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Therapy 1999; 6: 973–985.

    Article  CAS  Google Scholar 

  52. Ren C, Kumar S, Shaw DR, Ponnazhagan S . Genomic stability of self-complementary adeno-associated virus 2 during early stages of transduction in mouse muscle in vivo. Human Gene Ther 2005; 16: 1047–1057.

    Article  CAS  Google Scholar 

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Acknowledgements

Financial support of the National Institutes of Health Grants R01CA90850, R01CA98817, the US Army Department of Defense Grants BC044440, PC020372, PC050949 and the Susan G Komen Breast Cancer Foundation Grant BCTR50206 is gratefully acknowledged.

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  1. Department of Pathology, The University of Alabama at Birmingham, Birmingham, AL, USA

    T Isayeva, C Ren & S Ponnazhagan

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Correspondence to S Ponnazhagan.

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Isayeva, T., Ren, C. & Ponnazhagan, S. Intraperitoneal gene therapy by rAAV provides long-term survival against epithelial ovarian cancer independently of survivin pathway. Gene Ther 14, 138–146 (2007). https://doi.org/10.1038/sj.gt.3302853

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