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Chronic Myeloid Leukemia, BCR/ABL Studies and Myeloproliferative Disorders

Synergistic activity of imatinib and 17-AAG in imatinib-resistant CML cells overexpressing BCR-ABL – Inhibition of P-glycoprotein function by 17-AAG

Abstract

Overexpression of BCR-ABL and P-glycoprotein (Pgp) are two of the known mechanisms of imatinib resistance. As combination therapy may allow to overcome drug resistance, we investigated the effect of combination treatment with imatinib and 17-allylamino-17-demethoxygeldanamycin (17-AAG), a heat-shock protein 90 (Hsp90) inhibitor, on different imatinib-sensitive and imatinib-resistant CML cell lines. In imatinib-sensitive cells, combination index (CI) values obtained using the method of Chou and Talalay indicated additive (CI=1) or marginally antagonistic (CI>1) effects following simultaneous treatment with imatinib and 17-AAG. In imatinib-resistant cells both drugs acted synergistically (CI<1). In primary chronic-phase CML cells additive or synergistic effects of the combination of imatinib plus 17-AAG were discernible. Annexin V/propidium iodide staining showed that the activity of imatinib plus 17-AAG is mediated by apoptosis. Combination treatment with imatinib plus 17-AAG was more effective in reducing the BCR-ABL protein level than 17-AAG alone. Monotherapy with 17-AAG decreased P-glycoprotein activity, which may increase intracellular imatinib levels and contribute to the sensitization of CML cells to imatinib. The results suggest that combination of imatinib and 17-AAG may be useful to overcome imatinib resistance in a clinical setting.

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References

  1. Druker BJ, Tamura S, Buchdunger E, Ohno S, Segal GM, Fanning S et al. Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nat Med 1996; 2: 561–566.

    Article  CAS  PubMed  Google Scholar 

  2. Druker BJ, Talpaz M, Resta DJ, Peng B, Buchdunger E, Ford JM et al. Efficacy and safety of a specific inhibitor of the BCR-ABL tyrosine kinase in chronic myeloid leukemia. N Engl J Med 2001; 344: 1031–1037.

    Article  CAS  PubMed  Google Scholar 

  3. Druker BJ, Sawyers CL, Kantarjian H, Resta DJ, Reese SF, Ford JM et al. Activity of a specific inhibitor of the BCR-ABL tyrosine kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med 2001; 344: 1038–1042.

    Article  CAS  PubMed  Google Scholar 

  4. Gorre ME, Mohammed M, Ellwood K, Hsu N, Paquette R, Rao PN et al. Clinical resistance to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification. Science 2001; 293: 876–880.

    Article  CAS  PubMed  Google Scholar 

  5. Hochhaus A, Kreil S, Corbin AS, La Rosée P, Müller MC, Lahaye T et al. Molecular and chromosomal mechanisms of resistance to imatinib (STI571) therapy. Leukemia 2002; 16: 2190–2196.

    Article  CAS  PubMed  Google Scholar 

  6. von Bubnoff N, Schneller F, Peschel C, Duyster J . BCR-ABL gene mutations in relation to clinical resistance of Philadelphia-chromosome-positive leukaemia to STI571: a prospective study. Lancet 2002; 359: 487–491.

    Article  CAS  PubMed  Google Scholar 

  7. von Bubnoff N, Peschel C, Duyster J . Resistance of Philadelphia-chromosome positive leukemia towards the kinase inhibitor imatinib (STI571, Glivec): a targeted oncoprotein strikes back. Leukemia 2003; 17: 829–838.

    Article  CAS  PubMed  Google Scholar 

  8. Mahon FX, Deininger MW, Schultheis B, Chabrol J, Reiffers J, Goldman JM et al. Selection and characterization of BCR-ABL positive cell lines with differential sensitivity to the tyrosine kinase inhibitor ST571: diverse mechanisms of resistance. Blood 2000; 96: 1070–1079.

    CAS  PubMed  Google Scholar 

  9. Mahon FX, Belloc F, Lagarde V, Chollet C, Moreau-Gaudry F, Reiffers J et al. MDR1 gene overexpression confers resistance to imatinib mesylate in leukemia cell line models. Blood 2003; 101: 2368–2373.

    Article  CAS  PubMed  Google Scholar 

  10. Topaly J, Zeller WJ, Fruehauf S . Combination therapy with imatinib mesylate (STI571): synopsis of in vitro studies. Br J Haematol 2002; 119: 3–14.

    Article  CAS  PubMed  Google Scholar 

  11. Tipping AJ, Mahon FX, Zafirides G, Lagarde V, Goldman JM, Melo JV . Drug responses of imatinib mesylate-resistant cells: synergism of imatinib with other chemotherapeutic drugs. Leukemia 2002; 16: 2349–2357.

    Article  CAS  PubMed  Google Scholar 

  12. La Rosée P, O'Dwyer ME, Druker BJ . Insights from pre-clinical studies for new combination treatment regimens with the Bcr-Abl kinase inhibitor imatinib mesylate (Gleevec/Glivec) in chronic myelogenous leukemia: a translational perspective. Leukemia 2002; 16: 1213–1219.

    Article  PubMed  Google Scholar 

  13. Illmer T, Schaich M, Platzbecker U, Freiberg-Richter J, Oelschlagel U, von Bonin M et al. P-glycoprotein-mediated drug efflux is a resistance mechanism of chronic myelogenous leukemia cells to treatment with imatinib mesylate. Leukemia 2004; 18: 401–408.

    Article  CAS  PubMed  Google Scholar 

  14. Topaly J, Zeller WJ, Fruehauf S . Synergistic activity of the new ABL-specific tyrosine kinase inhibitor STI571 and chemotherapeutic drugs on BCR-ABL-positive chronic myelogenous leukemia cells. Leukemia 2001; 15: 342–347.

    Article  CAS  PubMed  Google Scholar 

  15. Chou TC, Talalay P . Quantitative analysis of dose–effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 1984; 22: 27–55.

    Article  CAS  PubMed  Google Scholar 

  16. Laemmli UK . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680–685.

    Article  CAS  PubMed  Google Scholar 

  17. Fruehauf S, Srbic K, Seggewiss R, Topaly J, Ho AD . Functional characterization of podia formation in normal and malignant hematopoietic cells. J Leukoc Biol 2002; 71: 425–432.

    CAS  PubMed  Google Scholar 

  18. Müller S, Neusser M, Wienberg J . Towards unlimited colors for fluorescence in situ hybridization (FISH). Chromosome Res 2002; 10: 223–232.

    Article  PubMed  Google Scholar 

  19. Speicher MR, Ballard GS, Ward DC . Karyotyping human chromosomes by combinatorial multi-fluor FISH. Nat Genet 1996; 12: 368–375.

    Article  CAS  PubMed  Google Scholar 

  20. Shtivelman E, Lifshitz B, Gale RP, Canaani E . Fused transcript of abl and bcr genes in chronic myelogenous leukaemia. Nature 1985; 315: 550–554.

    Article  CAS  PubMed  Google Scholar 

  21. Jerome V, Leger J, Devin J, Baulieu EE, Catelli MG . Growth factors acting via tyrosine kinase receptors induce HSP90 alpha gene expression. Growth Factors 1991; 4: 317–327.

    Article  CAS  PubMed  Google Scholar 

  22. Buchdunger E, Cioffi CL, Law N, Stover D, Ohno-Jones S, Druker BJ et al. Abl protein-tyrosine kinase inhibitor STI571 inhibits in vitro signal transduction mediated by c-kit and platelet-derived growth factor receptors. J Pharmacol Exp Ther 2000; 295: 139–145.

    CAS  PubMed  Google Scholar 

  23. Duhrsen U, Martinez T, Vohwinkel G, Ergun S, Sun L, McMahon G et al. Effects of vascular endothelial and platelet-derived growth factor receptor inhibitors on long-term cultures from normal human bone marrow. Growth Factors 2001; 19: 1–17.

    Article  CAS  PubMed  Google Scholar 

  24. Wang EJ, Casciano CN, Clement RP, Johnson WW . In vitro flow cytometry method to quantitatively assess inhibitors of P-glycoprotein. Drug Metab Dispos 2000; 28: 522–528.

    CAS  PubMed  Google Scholar 

  25. Sonneveld P, Wiemer E . Assays for the analysis of P-glycoprotein in acute myeloid leukemia and CD34 subsets of AML blasts. Leukemia 1997; 11: 1160–1165.

    Article  CAS  PubMed  Google Scholar 

  26. Weisberg E, Griffin JD . Mechanism of resistance to the ABL tyrosine kinase inhibitor STI571 in BCR/ABL-transformed hematopoietic cell lines. Blood 2000; 95: 3498–3505.

    CAS  PubMed  Google Scholar 

  27. Keeshan K, Mills KI, Cotter TG, McKenna SL . Elevated Bcr-Abl expression levels are sufficient for a haematopoietic cell line to acquire a drug-resistant phenotype. Leukemia 2001; 15: 1823–1833.

    Article  CAS  PubMed  Google Scholar 

  28. Tipping AJ, Mahon FX, Lagarde V, Goldman JM, Melo JV . Restoration of sensitivity to STI571 in STI571-resistant chronic myeloid leukemia cells. Blood 2001; 98: 3864–3867.

    Article  CAS  PubMed  Google Scholar 

  29. Schulte TW, Neckers LM . The benzoquinone ansamycin 17-allylamino-17-demethoxygeldanamycin binds to HSP90 and shares important biologic activities with geldanamycin. Cancer Chemother Pharmacol 1998; 42: 273–279.

    Article  CAS  PubMed  Google Scholar 

  30. Ochel HJ, Eichhorn K, Gademann G . Geldanamycin: The prototype of a class of antitumor drugs targeting the heat shock protein 90 family of molecular chaperones. Cell Stress Chaperones 2001; 6: 105–112.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. An WG, Schulte TW, Neckers LM . The heat shock protein 90 antagonist geldanamycin alters chaperone association with p210bcr-abl and v-src before their degradation by the proteasome. Cell Growth Differ 2000; 11: 355–360.

    CAS  PubMed  Google Scholar 

  32. Nimmanapalli R, O'Bryan E, Bhalla K . Geldanamycin and its analogue 17-allylamino-17-demethoxygeldanamycin lowers Bcr-Abl levels and induces apoptosis and differentiation of Bcr-Abl-positive human leukemic blasts. Cancer Res 2001; 61: 1799–1804.

    CAS  PubMed  Google Scholar 

  33. Nimmanapalli R, O'Bryan E, Huang M, Bali P, Burnette PK, Loughran T et al. Molecular characterization and sensitivity of STI-571 (imatinib mesylate, Gleevec)-resistant, Bcr-Abl-positive, human acute leukemia cells to SRC kinase inhibitor PD180970 and 17-allylamino-17-demethoxygeldanamycin. Cancer Res 2002; 62: 5761–5769.

    CAS  PubMed  Google Scholar 

  34. Gorre ME, Ellwood-Yen K, Chiosis G, Rosen N, Sawyers CL . BCR-ABL point mutants isolated from patients with imatinib mesylate-resistant chronic myeloid leukemia remain sensitive to inhibitors of the BCR-ABL chaperone heat shock protein 90. Blood 2002; 100: 3041–3044.

    Article  CAS  PubMed  Google Scholar 

  35. Banerji U, Judson I, Workman P . The clinical applications of heat shock protein inhibitors in cancer – present and future. Curr Cancer Drug Targets 2003; 3: 385–390.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Bernhard Berkus (German Cancer Research Center, Heidelberg, Germany), Brigitte Geisler and Carmen Hoppstock (both Department of Internal Medicine V, University of Heidelberg) for their excellent technical support, Carina Ittrich (German Cancer Research Center, Heidelberg, Germany) for the statistical calculations, Brigitte Schoell and Heidi Holtgreve-Grez (both Institute of Human Genetics, University of Heidelberg) for chromosome preparation and FISH experiments and Dr E Buchdunger (Novartis, Basel, Switzerland) for providing imatinib mesylate.

This work was supported in part by the Deutsche José Carreras Leukämie Stiftung (Grant DJCLS-R00/03 to SF and WJZ), by the competence network ‘Acute and Chronic Leukemias’ of the Federal Ministry of Education and Research (BMBF Grant 01GI9974 to AJ) and by the Medical Faculty of the University of Heidelberg (Juniorantrag to JT).

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Radujkovic, A., Schad, M., Topaly, J. et al. Synergistic activity of imatinib and 17-AAG in imatinib-resistant CML cells overexpressing BCR-ABL – Inhibition of P-glycoprotein function by 17-AAG. Leukemia 19, 1198–1206 (2005). https://doi.org/10.1038/sj.leu.2403764

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