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01-10-2008 | Original Article

Cytotoxic effects of treosulfan and busulfan against leukemic cells of pediatric patients

Authors: Doreen Munkelt, Ulrike Koehl, Stephan Kloess, Stefanie-Yvonne Zimmermann, Rabiá El Kalaäoui, Sibylle Wehner, Dirk Schwabe, Thomas Lehrnbecher, Ralf Schubert, Joerg Kreuter, Thomas Klingebiel, Ruth Esser

Published in: Cancer Chemotherapy and Pharmacology | Issue 5/2008

Abstract

Purpose

The alkylating agent treosulfan exerts a high cytotoxic activity against various malignant cells. Due to limited non-hematological toxicity, treosulfan might be a promising compound in myeloablative therapy for hematopoietic transplantation in children. Since in vitro data regarding the activity of treosulfan against childhood leukemic cells are limited, we compared the effect of treosulfan and busulfan against pediatric leukemic and non-malignant cells.

Experimental design

Both agents were tested alone and in combination with fludarabine by means of the MTT and/or a five color-flow cytometric assay. Moreover, the induction of apoptosis by treosulfan was investigated via regulation of the proteinase caspase 3.

Results

Treosulfan was more active against leukemic cells of 20 children as well as against 3 leukemia-derived cell lines than busulfan, with increasing IC₅₀ values from initial diagnosis to relapse. Overall purified stem cells were most sensitive, followed by CD56⁺CD3⁻ NK and CD3⁺ T cells. The combination of treosulfan with fludarabine resulted in a synergistic effect against leukemic cells. In malignant cells, treosulfan induced rapid cell apoptosis measured by the activation of the centrally proteinase caspase 3.

Conclusion

Our results indicate that treosulfan has activity against pediatric leukemic cells, myeloablative potential and immunosuppressive properties suitable for conditioning regimen in childhood malignancies.

Blaise D, Maraninchi D, Archimbaud E, Reiffers J, Devergie A, Jouet JP, Milpied N, Attal M, Michallet M, Ifrah N (1992) Allogeneic bone marrow transplantation for acute myeloid leukemia in first remission: a randomized trial of a busulfan-cytoxan versus cytotxan-total body irradiation as preparative regimen. Blood 79:2578–2582PubMed

Beck O, Seidl C, Lehrnbecher T, Kreyenberg H, Schwabe D, Klingebiel T, Seifried E, Bader P, Koehl U (2006) Quantification of chimerism within peripheral blood, bone marrow and purified leukocyte subsets: comparison of singleplex and multiplex PCR amplification of short tandem repeat (STR) loci. Eur J Haematol 76:237–244PubMedCrossRef

Beelen DW, Trenschel R, Casper J, Freund M, Hilger RA, Scheulen ME, Basara N, Fauser AA, Hertenstein B, Mylius HA, Baumgart J, Pichlmeier U, Hahn JR, Holler E (2005) Dose-escalated treosulphan in combination with cyclophosphamide as a new preparative regimen for allogeneic haematopoietic stem cell transplantation in patients with an increased risk for regimen-related complications. Bone Marrow Transplant 35:233–241PubMedCrossRef

Bene MC, Castoldi G, Knapp W, Ludwig WD, Matutes E, Orfao A, van´t Veer MB (1995) Proposals for the immunological classification of acute leukemias. European group for the immunological characterization of leukemias (EGIL). Leukemia 9:1783–1786PubMed

Bosanquet AG, Burlton AR (1994) Airborne cytotoxicity in the DiSC assay caused by solutions of treosulfan but not busulphan. Cytotechnology 16:131–136PubMedCrossRef

Casper J, Knauf W, Kiefer T, Wolff D, Steiner B, Hammer U, Wegener R, Kleine HD, Wilhelm S, Knopp A, Hartung G, Dölken G, Freund M (2004) Treosulfan and fludarabine: a new toxicity-reduced conditioning regimen for allogeneic haematopoietic stem cell transplantation. Blood 103:725–731PubMedCrossRef

Chou TC, Talalay P (1988) Quantitative analysis of dose-effect relationship: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55CrossRef

Den Boer M, Harms DO, Pieters R, Kazemier KM, Gobel U, Körholz D, Graubner U, Haas RJ, Jorch N, Spaar HJ, Kaspers GJ, Kamps WA, Van der Does-Van den Berg A, Van Wering ER, Veerman AJ, Janka-Schaub GE (2003) Patient stratification based on prednisolone-vincristine-asparaginase resistance profiles in children with acute lymphoblastic leukemia. J Clin Oncol 21:3262–3268CrossRef

Di Nicolantonio F, Knight LA, Di Palma S, Sharma S, Whitehouse PA, Mercer SJ, Charlton PA, Norris D, Cree IA (2004) Ex vivo characterization of XR11576 (MLN576) against ovarian cancer and other solid tumors. Anticancer Drugs 15:849–860PubMedCrossRef

10.

Drabko K, Zawitkowska-Klaczynska J, Wojcik B, Choma M, Zaucha-Prazmo A, Kowalczyk J, Gorczynska E, Toporski J, Kalwak K, Turkiewicz D, Chybicka A (2005) Megachemotherapy followed by autologous stem cell transplantation in children with Ewing’s sarcoma. Pediatr Transplant 9:618–621PubMedCrossRef

11.

Feyerabend S, Feil G, Krug J, Kassen A, Stenzl A (2007) Cytotoxic effects of treosulfan on prostate cancer cell lines. Anticancer Res 27:2403–2408PubMed

12.

Fichtner I, Becker M, Baumgart J (2003) Antileukaemic activity of treosulfan in xenografted human acute lymphoblastic leukaemias (ALL). Eur J Cancer 39:801–807PubMedCrossRef

13.

Gutiérrez MI, Siraj AK, Ibrahim MM, Hussain A, Bhatia K (2005) Childood and adult ALL: differences in epigenetic lesions associated with cell cycle genes. Am J Hematol 80:158–160PubMedCrossRef

14.

Hilger RA, Harstrick A, Eberhardt W, Oberhoff C, Skorzec M, Baumgart J, Seeber S, Scheulen ME (1998) Clinical pharmacokinetics of intravenous treosulfan in patients with advanced solid tumours. Cancer Chemother Pharmacol 42:99–104PubMedCrossRef

15.

Koehl U, Esser R, Zimmermann S, Tonn T, Kotchetkov R, Bartling T, Sörensen J, Grüttner HP, Bader P, Seifried E, Martin H, Lang P, Passweg J, Klingebiel T, Schwabe D (2005) Ex vivo expansion of highly purified NK-cells for immunotherapy after haploidentical stem cell transplantation in children. Klin Padiatr 217:345–350PubMedCrossRef

16.

Kopf- Maier P (1998) The alkylator treosulfan shows activity towards human renal-cell carcinoma in vivo and in vitro. In vivo 12:275–288PubMed

17.

Kröger N, Shimoni A, Zabelina T, Schieder H, Panse J, Ayuk F, Wolschke C, Renges H, Dahlke J, Atanackovic D, Nagler A, Zander A (2006) Reduced-toxicity conditioning with treosulfan, fludarabine and ATG as preparative regimen for allogeneic stem cell transplantation (alloSCT) in elderly patients with secondary acute myeloid leukemia (sAML) or myelodysplastic syndrome (MDS). Bone Marrow Transplant 37:339–344PubMedCrossRef

18.

Lanvers-Kaminsky C, Bremer A, Dirksen U, Jurgens H, Boos J (2006) Cytotoxicity of treosulfan and busulfan on pediatric tumor cell lines. Anticancer Drugs 17:657–662PubMedCrossRef

19.

Litzow MR, Prez WS, Klein JP, Bollwell BJ, Camitta B, Copelan EA, Gale RP, Giralt SA, Keating A, Lazarus HM, Marjus DI, McCarthy PL, Miller CB, Milone G, Prentice HG, Russel JA, Schultz KR, Trigg ME, Weisdorf DJ, Horowitz MM (2002) Comparison of outcome following allogeneic bone marrow transplantation with cyclophosphamide-total body irradiation versus busulfan-cyclophosphamide conditioning regimen for acute myelogenous leukaemia in first remission. Br J Haematol 119:1115–1124PubMedCrossRef

20.

Meinhardt G, Dayyani F, Jahrsdörfer B, Baumgart J, Emmerich B, Schmidmaier R (2003) Treosulfan is an effective inducer of cell death in myeloma cell lines and primary myeloma cells from patients. Br J Haematol 122: 892–899PubMedCrossRef

21.

Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63PubMedCrossRef

22.

Ploemacher RE, Johnson KW, Rombouts EJC, Kenol E, Westerhof GR, Baumgart J, White-Scharf ME, Down JD (2004) Addition of Treosulfan to a nonmyeloablative conditioning regimen results in enhanced chimerism and immunologic tolerance in an experimental allogeneic bone marrow transplant model. Biol Blood Marrow Transplant 10:236–245PubMedCrossRef

23.

Sanderson BJ, Johnson KJ, Henner WD (1991) Dose-dependent cytotoxic and mutagenic effects of antineoplastic alkylating agents on human lymphoblastoid cells. Environ Mol Mutagen 17:238–243PubMedCrossRef

24.

Sanyal U, Nanda R, Samanta S, Pain A, Dutta S, Verma AS, Rider BJ, Agrawal KC (2000) Evaluation of dimethylaminosulfonates of alkane diols as a novel group of anticancer agents. Cancer Lett 155:89–97PubMedCrossRef

25.

Sauer M, Zeidler C, Meissner B, Rehe K, Hanke A, Welte K, Lohse P, Sykora K-W (2007) Substitution of cyclophosphamide and busulfan by fludarabine, treosulfan and melphalan in a preparative regimen for children and adolescents with Shwachman-Diamond syndrome. Bone Marrow Transplant 39:143–147PubMedCrossRef

26.

Scheulen ME, Hilger RA, Oberhoff C, Casper J, Freund M, Josten KM, Bornhauser M, Ehninger G, Berdel WE, Baumgart J, Harstrick A, Bojko P, Wolf HH, Schindler AE, Seeber S (2000) Clinical phase I dose escalation and pharmacokinetic study of high-dose chemotherapy with treosulfan and autologous peripheral blood stem cell transplantation in patients with advanced malignancies. Clin Cancer Res 6:4209–4216PubMed

27.

Schmidmaier R, Oellerich M, Baumgart J, Emmerich B, Meinhardt G (2004) Treosulfan-induced apoptosis in acute myeloid leukemia cells is accompanied by translocation of protein kinase C delta and enhanced by bryostatin-1. Exp Hematol 32:76–96PubMedCrossRef

28.

Schmidmaier R, Baumann P, Emmerich B, Meinhardt G (2006) Evaluation of chemosensitivity of human bone marrow stromal cells—differences between common chemotherapeutic drugs. Anticaner Res 26:347–350

29.

Sjoo F, Hassan Z, Abedi-Valugerdi M, Griskevicius L, Nilsson C, Remberger M, Aschan J, Concha H, Gaughan U, Hassan M (2006) Myeloablative and immunosuppressive properties of treosulfan in mice. Exp Hematol 34:115–121PubMedCrossRef

30.

Styczynski J, Toporski J, Wysocki M, Debski R, Chybicka A, Boruczkowski D, Wachowiak J, Wojcik B, Kowalczyk J, Gil L, Balwierz W, Matysiak M, Krawczuk-Rybak M, Balcerska A, Sonta-Jakimczyk D (2007) Fludarabine, treosulfan and etoposide sensitivity and the outcome of hematopoietic stem cell transplantation in childhood acute myeloid leukemia. Anticancer Res 27:1547–1552PubMed

31.

Vassal G, Koscielny S, Challine D, Valteau-Couanet D, Boland I, Deroussent A, Lemerle J, Gouyette A, Hartmann O (1996) Busulfan disposition and hepatic veno-occlusive disease in children undergoing bone marrow transplantation. Cancer Chemother Pharmacol 37:247–253PubMedCrossRef

32.

Vergnon JM, Boucheron S, Riffat J, Guy C, Blanc P, Emonot A (1988) Interstitial pneumopathies caused by busulfan. Histologic, developmental and bronchoalveolar lavage analysis of 3 cases. Rev Med Interne 9:377–383PubMedCrossRef

33.

Werner S, Mendoza A, Hilger RA, Erlacher M, Reichardt W, Lissat A, Konanz C, Uhl M, Niemeyer CM, Khanna C, Kontny U (2007) Preclinical studies of treosulfan demonstrate potent activity in Ewing’s sarcoma. Cancer Chemother Pharmacol. (Epub ahead of print)

34.

Westerhof GR, Ploemacher RE, Boudewijn A, Blokland I, Dillingh JH, McGown AT, Hadfield JA, Dawson MJ, Down JD (2000) Comparison of different busulfan analogues for depletion of hematopoietic stem cells and promotion of donor-type chimerism in murine bone marrow transplant recipients. Cancer Res 60:5470–5478PubMed

35.

Zimmermann SY, Esser R, Rohrbach E, Klingebiel T, Koehl U (2005) A novel four-colour flowcytometric assay to determine natural killer cell or T cell-mediated cellular cytotoxicity against leukaemic cells in peripheral or bone marrow specimens containing greater than 20% of normal cells. J Immunol Methods 296:63–76PubMedCrossRef

Title: Cytotoxic effects of treosulfan and busulfan against leukemic cells of pediatric patients
Authors: Doreen Munkelt
Ulrike Koehl
Stephan Kloess
Stefanie-Yvonne Zimmermann
Rabiá El Kalaäoui
Sibylle Wehner
Dirk Schwabe
Thomas Lehrnbecher
Ralf Schubert
Joerg Kreuter
Thomas Klingebiel
Ruth Esser
Publication date: 01-10-2008
Publisher: Springer-Verlag
Published in: Cancer Chemotherapy and Pharmacology / Issue 5/2008
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-007-0669-3

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Purpose

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