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Reduced effect of gemtuzumab ozogamicin (CMA-676) on P-glycoprotein and/or CD34-positive leukemia cells and its restoration by multidrug resistance modifiers

Leukemia volume 16, pages 813–819 (2002)Cite this article

Abstract

Gemtuzumab ozogamicin (CMA-676), a calicheamicin-conjugated humanized anti-CD33 mouse monoclonal antibody, has recently been introduced clinically as a promising drug for the treatment of patients with acute myeloid leukemia (AML), more than 90% of which express CD33 antigen. However, our recent study suggested that CMA-676 was excreted by a multi- drug-resistance (MDR) mechanism in P-glycoprotein (P-gp)-expressing leukemia cell lines. We analyzed the in vitro effects of CMA-676 on leukemia cells from 27 AML patients in relation to the amount of P-gp, MDR-associated protein 1 (MRP1), CD33 and CD34, using a multi-laser-equipped flow cytometer. The cytocidal effect of CMA-676, estimated by the amount of hypodiploid portion on cell cycle, was inversely related to the amount of P-gp estimated by MRK16 monoclonal antibody (P = 0.004), and to the P-gp function assessed by intracellular rhodamine-123 accumulation in the presence of PSC833 or MS209 as a MDR modifier (P = 0.0004 and P = 0.002, respectively). In addition, these MDR modifiers reversed CMA-676 resistance in P-gp-expressing CD33+ leukemia cells (P = 0.001 with PSC833 and P = 0.0007 with MS209). In CD33+ AML cells from 13 patients, CMA-676 was less effective on CD33+CD34+ than CD33+CD34− cells (P = 0.002). PSC833 partially restored the effect of CMA-676 in CD33+CD34+ cells. These results suggest that the combined use of CMA-676 and a MDR modifier will be more effective on CD33+ AML with P-gp-related MDR.

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References

  1. Pirker R, Wallner J, Geissler K, Linkesch W, Haas OA, Bettelheim P, Hopfner M, Scherrer R, Valent P, Havelec L, Ludwig H, Lechner K . MDR1 gene expression and treatment outcome in acute myeloid leukemia J Natl Cancer Inst 1991 83: 708–712

    Article  CAS  Google Scholar 

  2. Campos L, Guyotat D, Archimbaud E, Oriol PC, Tsuruo T, Troncy J, Treille D, Fiere D . Clinical significance of multidrug resistance P-glycoprotein expression on acute nonlymphoblastic leukemia cells at diagnosis Blood 1992 79: 473–476

    CAS  PubMed  Google Scholar 

  3. Michieli M, Damiani D, Geromin A, Michelutti A, Fanin R, Raspadori D, Russo D, Visani G, Dinota A, Pileri S, Tsuruo T, Grandi M, Baccarani M, Tura S . Overexpression of multidrug resistance-associated p170-glycoprotein in acute non-lymphocytic leukemia Eur J Haematol 1992 48: 87–92

    Article  CAS  Google Scholar 

  4. te Boekhorst PAW, de Leeuw K, Shoester M, Wittebol S, Nooter K, Hagemeijer A, Lowenberg B, Sonneveld P . Predominance of functional multidrug resistance (MDR-1) phenotype in CD34+ acute myeloid leukemia cells Blood 1993 82: 3157–3162

    CAS  PubMed  Google Scholar 

  5. Sonneveld P, van Dongen JJ, Hagemeijer A, van Lom K, Nooter K, Shoester M, Adriaansen HJ, Tsuruo T, de Leeuw K . High expression of the multidrug resistance P-glycoprotein in high-risk myelodysplasia is associated with immature phenotype Leukemia 1993 7: 963–969

    CAS  PubMed  Google Scholar 

  6. Takeshita A, Shinjo K, Ohnishi K, Ohno R . Expression of multidrug resistance P-glycoprotein in myeloid progenitor cells of different phenotype: comparison between normal bone marrow cells and leukaemia cells Br J Haematol 1996 93: 18–21

    Article  CAS  Google Scholar 

  7. Merlin JL, Guerci AP, Marchal S, Bour C, Colosetti P, Kataki A, Guerci O . Influence of SDZ-PSC833 on daunorubicin intracellular accumulation in bone marrow specimens from patients with acute myeloid leukaemia Br J Haematol 1998 103: 480–487

    Article  CAS  Google Scholar 

  8. Sato W, Fukazawa N, Nakanishi O, Baba M, Suzuki T, Yano O, Naito M, Tsuruo T . Reversal of multidrug resistance by a novel quinoline derivative, MS-209 Cancer Chemother Pharmacol 1995 35: 271–277

    Article  CAS  Google Scholar 

  9. Multani PS, Grossbard ML . Monoclonal antibody-based therapies for hematologic malignancies J Clin Oncol 1998 16: 3691–3710

    Article  CAS  Google Scholar 

  10. Freeman SD, Kelm S, Barber EK, Crocker PR . Characterization of CD33 as a new member of the sialoadhesin family of cellular interaction molecules Blood 1995 85: 2005–2012

    CAS  PubMed  Google Scholar 

  11. Wagner JE, Collins D, Fuller S, Schain LR, Berson AE, Almici C, Hall MA, Chen KE, Okarma TB, Lebkowski JS . Isolation of small, primitive human hematopoietic stem cells: distribution of cell surface cytokine receptors and growth in SCID-Hu mice Blood 1995 86: 512–523

    CAS  PubMed  Google Scholar 

  12. Sheinberg DA, Lovett D, Divgi CR, Graham MC, Berman E, Pentlow K, Feirt N, Finn RD, Clarkson BD, Gee TS, Larson SM, Oettgen HF, Old LJ . A phase I trial of monoclonal antibody M195 in acute myelogenous leukemia: specific bone marrow targeting and internalization of radionuclide J Clin Oncol 1991 9: 478–490

    Article  Google Scholar 

  13. Caron PC, Co MS, Bull MK, Avdalovic NM, Queen C, Scheinberg DA . Biological and immunological features of Humanized M195 (Anti-CD33) monoclonal antibodies Cancer Res 1992 52: 6761–6767

    CAS  PubMed  Google Scholar 

  14. Appelbaum FR, Matthews DC, Eary JF, Badger CC, Kellogg M, Press OW, Martin PJ, Fisher DR, Nelp WB, Thomas ED, Bernstein ID . The use of radiolabeled anti-CD33 antibody to augment marrow irradiation prior to marrow transplantation for acute myelogenous leukemia Transplantation 1992 54: 829–833

    Article  CAS  Google Scholar 

  15. Linenberger ML, Hong T, Flowers D, Sievers EL, Gooley TA, Bennett JM, Berger MS, Leopold LH, Appelbaum FR, Bernstein ID . Multidrug-resistance phenotype and clinical responses to gemtuzumab ozogamicin Blood 2001 98: 988–994

    Article  CAS  Google Scholar 

  16. Lee MD, Dunne TS, Siegel MM, Chang CC, Morton GO, Borders DB . Calichemicins, a novel family of antitumor antibiotics. 1. Chemistry and partial structure of calichemicin γ1I J Am Chem Soc 1987 109: 3464–3466

    Article  CAS  Google Scholar 

  17. Lee MD, Dunne TS, Chang CC, Ellestad GA, Siegel MM, Morton GO, McGahren WJ, Borders DB . Calichemicins, a novel family of antitumor antibiotics. 2. Chemistry and structure of calichemicin γ1I J Am Chem Soc 1987 109: 3466–3468

    Article  CAS  Google Scholar 

  18. Zein N, Poncin M, Nilakantan R, Ellestad GA . Calicheamicin γ1I and DNA: molecular recognition process responsible for site-specificity Science 1989 244: 697–699

    Article  CAS  Google Scholar 

  19. Hangeland JJ, De Voss JJ, Heath JA, Townsend CA . Specific abstraction of the 5′(S)-and 4′-deoxyribosyl hydrogen atoms from DNA by calicheamicin γ1I J Am Chem Soc 1992 114: 9200–9202

    Article  CAS  Google Scholar 

  20. Naito K, Takeshita A, Shigeno K, Nakamura S, Fujisawa S, Shinjo K, Yoshida H, Ohnishi K, Mori M, Terakawa S, Ohno R . Calicheamicin-conjugated humanized anti-CD33 monoclonal antibody (gemtuzumab zogamicin, CMA-676) shows cytocidal effect on CD33-positive leukemia cell lines, but is inactive on P-glycoprotein-expressing sublines Leukemia 2000 14: 1436–1443

    Article  CAS  Google Scholar 

  21. Matsui H, Takeshita A, Naito K, Shinjo K, Ohnishi K, Ohno R . Reduced effect of gemtuzumab ozogamicin (CMA-676) on P-glycoprotein and/or CD34 positive leukemia cells and its restoration by multidrug resistance modifiers Blood 2000 96 (Suppl.): 117a (Abstr.)

    Google Scholar 

  22. Leith CP, Kopecky KJ, Godwin J, McConnell T, Slovak ML, Chen IM, Head DR, Appelbaum FR, Willman CL . Acute myeloid leukemia in the elderly: assessment of multidrug resistance (MDR1) and cytogenetics distinguishes biologic subgroups with remarkably distinct responses to standard chemotherapy. A Southwest Oncology Group study Blood 1997 89: 3323–3329

    CAS  PubMed  Google Scholar 

  23. Bennett JM, Catovsky D, Daniel MT, Flandrin G, Galton DAG, Gralnick HR, Sultan C . Criteria for the diagnosis of acute leukemia of megakaryocyte lineage (M7) Ann Intern Med 1985 103: 460–462

    Article  CAS  Google Scholar 

  24. Bennett JM, Catovsky D, Daniel MT, Flandrin G, Galton DAG, Gralnick HR, Sultan C . Proposed revised criteria for the classification of acute myeloid leukemia Ann Intern Med 1985 103: 626–629

    Article  Google Scholar 

  25. Lacombe F, Durrieu F, Briais A, Dumain P, Belloc F, Bascans E, Reiffers J, Boisseau MR, Bernard PH . Flow cytometry CD45 gating for immunophenotyping of acute myeloid leukemia Leukemia 1997 11: 1878–1886

    Article  CAS  Google Scholar 

  26. Takeshita A, Shinjo K, Ohnishi K, Ohno R . New flow cytometric method for detection of minimally expressed multidrug resistance P-glycoprotein on normal and acute leukemia cells using biotinylated MRK16 and streptavidin-RED670 conjugate Jpn J Cancer Res 1995 86: 607–615

    Article  CAS  Google Scholar 

  27. den Boer ML, Zwaan CM, Pieters R, Kazemier KM, Rottier MMA, Flens MJ, Scheper RJ, Veerman AJP . Optimal immunocytochemical and flow cytometric detection of P-gp, MRP and LRP inchildhood acute lymphoblastic leukemia Leukemia 1997 11: 1078–1085

    Article  CAS  Google Scholar 

  28. Overton WR . Modified histogram subtraction technique for analysis of flow cytometry data Cytometry 1988 9: 619–626

    Article  CAS  Google Scholar 

  29. Muller MR, Lennartz K, Nowrousian MR, Dux R, Tsuruo T, Rajewsky MF, Seeber S . Improved flow-cytometric detection of lowP-glycoprotein expression in leukaemic blasts by histogramsubtraction analysis Cytometry 1994 15: 64–72

    Article  CAS  Google Scholar 

  30. Lamy T, Drenou B, Grulois I, Fardel O, Jacquelinet C, Goasguen J, Dauriac C, Amiot L, Bernard M, Fauchet R, Le Prise PY . Multi-drug resistance (MDR) activity in acute leukemia determined by rhodamine 123 efflux assay Leukemia 1995 9: 1549–1555

    CAS  PubMed  Google Scholar 

  31. Legrand O, Simonin G, Perrot JY, Zittoun R, Marie JP . Pgp and MRP activities using calcein-AM are prognostic factors in adult acute myeloid leukemia patients Blood 1998 91: 4480–4488

    CAS  PubMed  Google Scholar 

  32. Advani R, Saba HI, Tallman MS, Rowe JM, Wiernik PH, Ramek J, Dugan K, Lum B, Villena J, Davis E, Paietta E, Litchman M, Sikic BI, Greenberg PL . Treatment of refractory and relapsed acute myelogenous leukemia with combination chemotherapy plus the multidrug resistance modulater PSC833 (Valspodar) Blood 1999 93: 787–795

    CAS  PubMed  Google Scholar 

  33. Borg AG, Burgess R, Green LM, Scheper RJ, Liu Yin JA . P-glycoprotein and multidrug resistance-associated protein, but not lung resistance protein, lower the intracellular daunorubicin accumulation in acute myeloid leukaemia cells Br J Haematol 2000 108: 48–54

    Article  CAS  Google Scholar 

  34. Sievers EL, Larson RA, Stadtmauer EA, Estey E, Lowenberg B, Dombret H, Karanes C, Theobald M, Bennett JM, Sherman ML, Berger MS, Eten CB, Loken MR, van Dongen JJ, Bernstein ID, Appelbaum FR Mylotarg Study Group. Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukemia in first relapse J Clin Oncol 2001 19: 3244–3254

    Article  CAS  Google Scholar 

  35. 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  Google Scholar 

  36. Krause DS, Fackler MJ, Civin CI, May WS . CD34: structure, biology, and clinical utility Blood 1996 87: 1–13

    CAS  PubMed  Google Scholar 

  37. Osawa M, Hanada K, Hamada H, Nakauchi H . Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell Science 1996 273: 242–245

    Article  CAS  Google Scholar 

  38. Yin AH, Miraglia S, Zanjani ED, Almeida-Porada G, Ogawa M, Leary AG, Olweus J, Kearney J, Buck DW . AC133, a novel marker for human hematopoietic stem and progenitor cells Blood 1997 90: 5002–5012

    CAS  Google Scholar 

  39. Feuring-Buske M, Hogge DE . Hoechst 33342 efflux identifies a subpopulation of cytogenetically normal CD34(+)CD38(−) progenitor cells from patients with acute myeloid leukemia Blood 2001 97: 3882–3889

    Article  CAS  Google Scholar 

  40. Wulf GG, Wang RY, Kuehnle I, Weidner D, Marini F, Brenner MK, Andreeff M, Goodell MA . A leukemic stem cell with intrinsic drug efflux capacity in acute myeloid leukemia Blood 2001 98: 1166–1173

    Article  CAS  Google Scholar 

  41. Campos L, Rouault JP, Sabido O, Oriol P, Roubi N, Vasselon C, Archimbaud E, Magaud JP, Guyotat D . High expression of bcl-2 protein in acute myeloid leukemia cells is associated with poor response to chemotherapy Blood 1993 81: 3091–3096

    CAS  PubMed  Google Scholar 

  42. Rochlitz C, Lohri A, Bacchi M, Schmidt M, Nagel S, Fopp M, Fey MF, Herrmann R, Neubauer A . Axl expression is associated with adverse prognosis and with expression of Bcl-2 and CD34 in de novo acute myeloid leukemia (AML): results from a multicenter trial of the Swiss Group for Clinical Cancer Research (SAKK) Leukemia 1999 13: 1352–1358

    Article  CAS  Google Scholar 

  43. Michieli M, Damiani D, Ermacora A, Masolini P, Raspadori D, Visani G, Scheper RJ, Baccarani M . P-glycoprotein, lung resistance-related protein and multidrug resistance associated protein in de novo acute non-lymphocytic leukaemias: biological and clinical implications Br J Haematol 1999 104: 328–335

    Article  CAS  Google Scholar 

  44. Ross DD, Karp JE, Chen TT, Doyle LA . Expression of breast cancer resistance protein in blast cells from patients with acute leukemia Blood 2000 96: 365–368

    CAS  Google Scholar 

  45. Paietta E, Anderson J, Gallagher R, Bennett J, Yunis J, Cassileth P, Rowe J, Wiernik PH . The immunophenotype of acute promyelocytic leukemia (APL): an ECOG study Leukemia 1994 8: 1108–1112

    CAS  PubMed  Google Scholar 

  46. Guglielmi C, Martelli MP, Diverio D, Fenu S, Vegna ML, Cantu-Rajnoldi A, Biondi A, Cocito MG, Del Vecchio L, Tabilio A, Avvisati G, Basso G, Lo Coco F . Immunophenotype of adult andchildhood acute promyelocytic leukaemia: correlation withmorphology, type of PML gene breakpoint and clinical outcome. A cooperative Italian study on 196 cases Br J Haematol 1998 102: 1035–1041

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to express our sincere gratitude to Ms Yoshimi Suzuki, Ms Noriko Anma and Ms Keiko Ono for technical assistance. We are indebted to Wyeth-Ayerst Research Division of American Home Products for the supply of test substances.

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Authors and Affiliations

  1. Division of Hematology, Department of Medicine III, Hamamatsu University School of Medicine, Hamamatsu, Japan

    H Matsui, K Naito, K Shinjo, K Shigeno & K Ohnishi

  2. Department of Laboratory Medicine, Hamamatsu University School of Medicine, Hamamatsu, Japan

    A Takeshita & M Maekawa

  3. Equipment Center, Nagoya University, Nagoya, Japan

    Y Yamakawa

  4. Department of Internal Medicine II, Okayama University, Okayama, Japan

    M Tanimoto

  5. Division of Hematology, Hamamatsu Medical Center, Hamamatsu, Japan

    M Kobayashi

  6. Aichi Cancer Center, Nagoya, Japan

    R Ohno

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Matsui, H., Takeshita, A., Naito, K. et al. Reduced effect of gemtuzumab ozogamicin (CMA-676) on P-glycoprotein and/or CD34-positive leukemia cells and its restoration by multidrug resistance modifiers. Leukemia 16, 813–819 (2002). https://doi.org/10.1038/sj.leu.2402459

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