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A novel antisense inhibitor of MMP-9 attenuates angiogenesis, human prostate cancer cell invasion and tumorigenicity

Abstract

Androgen deprivation therapy causes a paradoxical elevation of matrix metalloproteinases (MMPs) including MMP-9 resulting in aggressive tumor phenotype in many patients with prostate cancer. In this study, we have evaluated a novel antisense phosphorodiamidate Morpholino oligomer (PMO) targeted against MMP-9 in models of angiogenesis and in human prostate xenograft in athymic mice. The treatment of androgen-independent DU145 human prostate cells with a 21-mer MMP-9 antisense PMO caused a dose-dependent inhibition of cell proliferation compared to scrambled or MMP-2 antisense PMO at similar concentrations. This was associated with decreases in MMP-9 expression, gelatinolytic activity and increased stability of the insulin-like growth factor-binding protein (IGFBP-3), a proapoptotic factor and MMP-9 substrate. In vitro invasion assays revealed a 40–60% inhibition of DU145 cell invasion in the presence of 25 μM MMP-9 antisense PMO. A significant decrease in endothelial cell migration and vascularization was observed in the Matrigel plug assay in mice when treated intraperitoneally with 300 μg/day MMP-9 antisense for 21 days. In the highly vascular DU145 tumor xenografts, MMP-9 inhibition caused decreased tumor growth with regression in 50% of the animals. Histological analysis revealed increased apoptosis and fibrous tissue deposits in the MMP-9 antisense-treated tumors compared to the scrambled and saline controls. No apparent toxicity or mortality was associated with the MMP-9 PMO treatment. In summary, the MMP-9 antisense PMO inhibited in vitro prostate cancer cell proliferation, invasion and in vivo angiogenesis. These data establish the feasibility of developing a site-directed, nontoxic antisense therapeutic agent for inhibiting local invasion and metastasis.

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References

  1. Greenlee RT, Hill-Harmon MB, Murray T, Thun M . Cancer statistics (2001). CA: Cancer J Clin. 2001;51:15–36.

    CAS  Google Scholar 

  2. Richie JP . Anti-androgens and other hormonal therapies for prostate cancer. Urology. 1999;54:15–18.

    Article  CAS  Google Scholar 

  3. Devi GR . Prostate cancer: status of current treatments and emerging antisense-based therapies. Curr Opin Mol Ther. 2002;4:138–148.

    CAS  PubMed  Google Scholar 

  4. Sadar MD, Hussain M, Bruchovsky N . Prostate cancer: molecular biology of early progression to androgen in-dependence. Endocrin Rel Cancer. 1999;6:487–502.

    Article  CAS  Google Scholar 

  5. Overall CM . Molecular determinants of metalloproteinase substrate specificity: matrix metalloproteinase substrate binding domains, modules, and exosites. Mol Biotechnol. 2002;22:51–86.

    Article  CAS  Google Scholar 

  6. Creemers EE, Cleutjens JP, Smits JF, Daemen MJ . Matrix metalloproteinase inhibition after myocardial infarction: a new approach to prevent heart failure? Circ Res. 2001;89:201–210 (Review).

    Article  CAS  Google Scholar 

  7. Porter KE, Turner NA . Statins for the prevention of vein graft stenosis: a role for inhibition of matrix metalloproteinase-9. Biochem Soc Trans. 2002;30:120–126.

    Article  CAS  Google Scholar 

  8. Lijnen HR . Plasmin and matrix metalloproteinases in vascular remodeling. Thromb Haemost. 2001;86:324–333. (Review).

    Article  CAS  Google Scholar 

  9. Harada H, Furuya M, Ishikura H, Shindo J, Koyanagi T, Yoshiki T . Expression of matrix metalloproteinase in the fluids of renal cystic lesions. J Urol. 2002;168:19–22.

    Article  Google Scholar 

  10. Faber-Elmann A, Sthoeger Z, Tcherniack A, Dayan M, Mozes E . Activity of matrix metalloproteinase-9 is elevated in sera of patients with systemic lupus erythematosus. Clin Exp Immunol. 2002;127:393–398.

    Article  CAS  Google Scholar 

  11. Giannelli G, Antonaci S . Gelatinases and their inhibitors in tumor metastasis: from biological research to medical applications. Histol Histopathol. 2002;17:339–345.

    CAS  PubMed  Google Scholar 

  12. Egeblad M, Werb Z . New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2002;2:161–174.

    Article  CAS  Google Scholar 

  13. Pajouh MS, Nagle RB, Breathnach R, Finch JS, Brawer MK, Bowden GT . Expression of metalloproteinase genes in human prostate cancer. J Cancer Res Clin Oncol. 1991;117:144–150.

    Article  CAS  Google Scholar 

  14. Lokeshwar BL, Selzer MG, Block NL, Gunja-Smith Z . Secretion of matrix metalloproteinases and their inhibitors (tissue inhibitor of metalloproteinases) by human prostate in explant cultures: reduced tissue inhibitor of metalloproteinase secretion by malignant tissues. Cancer Res. 1993;53:4493–4498.

    CAS  PubMed  Google Scholar 

  15. Stearns ME, Wang M . Type IV collagenase (M(r) 72,000) expression in human prostate: benign and malignant tissue. Cancer Res. 1993;53:878–883.

    CAS  PubMed  Google Scholar 

  16. Sternlicht MD, Werb Z . How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol. 2001;17:463–516.

    Article  CAS  Google Scholar 

  17. Rajah R, Valentinis B, Cohen P . IGFBP-3 induces apoptosis and mediates the effects of transforming growth factor-b1 on programmed cell death through a p53- and IGF-independent mechanism. J Biol Chem. 1997;272:12181–12188.

    Article  CAS  Google Scholar 

  18. Devi GR, Sprenger C, Graham D, Plymate S, Rosenfeld RG . Overexpression of IGFBP-3 induces cell growth inhibition and early apoptosis in malignant human prostate cancer cells. Prostate 2002;53:200–210.

    Article  CAS  Google Scholar 

  19. Cohen P, Peehl D, Graves H, Rosenfeld R . Biological effects of prostate specific antigen (PSA) as an IGF binding protein-3 (IGFBP-3) protease. J Endocrinol. 1994;142:407–415.

    Article  CAS  Google Scholar 

  20. Smith GL, Doherty AP, Mitchell H, Hanham IW, Christmas TJ, Epstein RJ . Inverse relation between prostate-specific antigen and insulin-like growth factor-binding protein 3 in bone metastases and serum of patients with prostate cancer. Lancet. 1999;354:2053–2054.

    Article  CAS  Google Scholar 

  21. Shariat SF, Lamb DJ, Kattan MW et al. Association of preoperative plasma levels of insulin-like growth factor 1 and insulin-like growth factor binding proteins-2 and -3 with prostate cancer invasion, progression, and metastasis. J Clin Oncol. 2002;20:833–841.

    Article  CAS  Google Scholar 

  22. Zucker S, Cao J, Chen WT . Critical appraisal of the use of matrix metalloproteinase inhibitors in cancer treatment. Oncogene. 2000;19:6642–6650.

    Article  CAS  Google Scholar 

  23. Overall CM, Lopez-Otin C . Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nat Rev Cancer. 2002;2:657–672.

    Article  CAS  Google Scholar 

  24. Summerton J, Weller D . Morpholino antisense oligomers: design, preparation, and properties. Antisense Nucleic Acids Drug Dev. 1997;7:187–195.

    Article  CAS  Google Scholar 

  25. Hudziak RM, Summerton J, Weller DD, Iversen PL . Antiproliferative effects of steric blocking phosphorodiamidate morpholino antisense agents directed against c-myc. Antisense Nucleic Acids Drug Dev. 2000;10:163–176.

    Article  CAS  Google Scholar 

  26. Devi GR, Oldenkamp JR, London CA, Iversen PL . Inhibition of human chorionic gonadotropin beta subunit modulates the mitogenic effect of c-myc in human prostate cancer cells. Prostate 2002;53:200–210.

    Article  CAS  Google Scholar 

  27. Attiga FA, Fernandez PM, Weeraratna AT, Manyak MJ, Patierno SR . Inhibitors of prostaglandin synthesis inhibit human prostate tumor cell invasiveness and reduce the release of matrix metalloproteinases. Cancer Res. 2000;60:4629–4637.

    CAS  PubMed  Google Scholar 

  28. Smith JP, Verderame MF, Zagon IS . Antisense oligonucleotides to gastrin inhibit growth of human pancreatic cancer. Cancer Lett. 1999;135:107–112.

    Article  CAS  Google Scholar 

  29. Sauter ER, Takemoto R, Litwin S, Herlyn M . p53 alone or in combination with antisense cyclin D1 induces apoptosis and reduces tumor size in human melanoma. Cancer Gene Ther. 2002;9:807–812.

    Article  CAS  Google Scholar 

  30. Iversen PL, Arora V, Acker AJ, Mason D, Devi GR . Efficacy of antisense Morpholino oligomer targeted to c-myc in prostate cancer xenograft murine model and a phase I safety study in humans. Clin Cancer Res. 2003;9:2510–2519.

    CAS  PubMed  Google Scholar 

  31. Desai SB, Libutti SK . Tumor angiogenesis and endothelial cell modulatory factors. J Immunother. 1999;22:186–211.

    Article  CAS  Google Scholar 

  32. Folkman J . Angiogenesis-dependent diseases. Semin Oncol. 2001;28:536–542.

    Article  CAS  Google Scholar 

  33. Lokeshwar BL . MMP inhibition in prostate cancer. Ann NY Acad Sci. 1999;878:271–289 (Review).

    Article  CAS  Google Scholar 

  34. Manes S, Llorente M, Lacalle RA, Gomez-Mouton C, Kremer L, Mira E, Martinez- AC . The matrix metalloproteinase-9 regulates the insulin-like growth factor-triggered autocrine response in DU-145 carcinoma cells. J Biol Chem. 1999;274:6935–6945.

    Article  CAS  Google Scholar 

  35. Angelloz-Nicoud P, Binoux M . Autocrine regulation of cell proliferation by the insulin-like growth factor (IGF) and IGF binding protein-3 protease system in a human prostate carcinoma cell line (PC-3). Endocrinology. 1995;136:5485–5492.

    Article  CAS  Google Scholar 

  36. Devi GR, Yang DY, Rosenfeld RG, Oh Y . Differential effects of IGFBP-3 and its proteolytic fragments on ligand binding, IGFIR signaling and cell surface association. Endocrinology. 2000;141:4171–4179.

    Article  CAS  Google Scholar 

  37. Devi GR, Oh Y, Rosenfeld RG . Effect of IGFBP-3 on IGF- and IGF-analog-induced IGFIR signaling. Growth Horm. IGF Res. 2001;11:231–239.

    Article  CAS  Google Scholar 

  38. Summerton J, Weller D . Antisense properties of morpholino oligomers. Nucleosides Nucleotides. 1997;16:889–898.

    Article  CAS  Google Scholar 

  39. Summerton J . Morpholino antisense oligomers: the case for an RNase H-independent structural type. Biochim Biophys Acta. 1999;1489:141–158.

    Article  CAS  Google Scholar 

  40. Iversen PL . PMO: favorable properties for sequence-specific inactivation. Curr Opin Mol Ther. 2001;3:235–238.

    CAS  PubMed  Google Scholar 

  41. Giles RV, Spiller DG, Clark RE, Tidd DM . Antisense morpholino oligonucleotide analog induces missplicing of c-myc mRNA. Antisense Nucleic Acids Drug Dev. 1999;9:213–220.

    Article  CAS  Google Scholar 

  42. Ghosh C, Stein D, Weller D, Iversen PL . Evaluation of antisense mechanisms of action. Methods Enzymol. 2000;313:135.

    Article  CAS  Google Scholar 

  43. Knapp DC, Mata JE, Reddy MT, Devi GR, Iversen PL . Resistance to chemotherapeutic drugs overcome by c-Myc inhibition in Lewis lung carcinoma murine model. Anti-Cancer Drugs. 2003;14:39–47.

    Article  CAS  Google Scholar 

  44. Arora V, Knapp DC, Reddy MT, Weller DD, Iversen PL . Bioavailability and efficacy of antisense morpholino oligomers targeted to c-Myc and cytochrome P-450 3A2 following oral administration in rats. J Pharm Sci. 2002;91:1009–1017.

    Article  CAS  Google Scholar 

  45. Arora V, Hannah TL, Iversen PL, Brand R . Transdermal use of phosphorodiamidate Morpholino oligomer AVI-4472 inhibits cytochrome P450 3A2 activity in male rats. Pharm Res. 2002;19:1465–1470.

    Article  CAS  Google Scholar 

  46. Henry SP, Novotny W, Leeds J, Auletta C, Kornbrust DJ . Inhibition of coagulation by a phosphorothioate oligonucleotide. Antisense Nucleic Acids Drug Dev. 1997;7:503–510.

    Article  CAS  Google Scholar 

  47. Iversen PL, Cornish KG, Iversen LJ, Mata JE, Bylund DB . Bolus intravenous injection of phosphorothioate oligonucleotides causes hypotension by acting as alpha1-adrenergic receptor antagonists. Toxicol Appl Pharm. 1999;160:289–296.

    Article  CAS  Google Scholar 

  48. Levin AA . A review of the issues in the pharmacokinetics and toxicology of phosphorothioate antisense oligonucleotides. Biochem Biophys Acta. 1999;1489:69–84.

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by a grant from Department of Defense-DAMD17-01-1-0017 (GRD) and funds from AVI BioPharma. We thank Melissa Cate and Andrew Kroecker for technical assistance; Dr Ojerio (Oregon State University) for expert advice on animal care issues; and Anita Sonn (Oregon State University) for assistance in preparation of slides. We also acknowledge the technical assistance provided by the synthesis and purification group at AVI BioPharma.

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Correspondence to Gayathri R Devi.

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London, C., Sekhon, H., Arora, V. et al. A novel antisense inhibitor of MMP-9 attenuates angiogenesis, human prostate cancer cell invasion and tumorigenicity. Cancer Gene Ther 10, 823–832 (2003). https://doi.org/10.1038/sj.cgt.7700642

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