Advertisement

Beta-irradiation used for systemic radioimmunotherapy induces apoptosis and activates apoptosis pathways in leukaemia cells

  • Claudia Friesen
  • Annelie Lubatschofski
  • Jörg Kotzerke
  • Inga Buchmann
  • Sven N. Reske
  • Klaus-Michael DebatinEmail author
Original Article

Abstract

Beta-irradiation used for systemic radioimmunotherapy (RIT) is a promising treatment approach for high-risk leukaemia and lymphoma. In bone marrow-selective radioimmunotherapy, beta-irradiation is applied using iodine-131, yttrium-90 or rhenium-188 labelled radioimmunoconjugates. However, the mechanisms by which beta-irradiation induces cell death are not understood at the molecular level. Here, we report that beta-irradiation induced apoptosis and activated apoptosis pathways in leukaemia cells depending on doses, time points and dose rates. After beta-irradiation, upregulation of CD95 ligand and CD95 receptor was detected and activation of caspases resulting in apoptosis was found. These effects were completely blocked by the broad-range caspase inhibitor zVAD-fmk. In addition, irradiation-mediated mitochondrial damage resulted in perturbation of mitochondrial membrane potential, caspase-9 activation and cytochrome c release. Bax, a death-promoting protein, was upregulated and Bcl-xL, a death-inhibiting protein, was downregulated. We also found higher apoptosis rates and earlier activation of apoptosis pathways after gamma-irradiation in comparison to beta-irradiation at the same dose rate. Furthermore, irradiation-resistant cells were cross-resistant to CD95 and CD95-resistant cells were cross-resistant to irradiation, indicating that CD95 and irradiation used, at least in part, identical effector pathways. These findings demonstrate that beta-irradiation induces apoptosis and activates apoptosis pathways in leukaemia cells using both mitochondrial and death receptor pathways. Understanding the timing, sequence and molecular pathways of beta-irradiation-mediated apoptosis may allow rational adjustment of chemo- and radiotherapeutic strategies.

Keywords

Radioimmunotherapy Beta-irradiation Apoptosis Leukaemia Rhenium-188 

Notes

Acknowledgements

We thank Stefanie Rath for excellent technical assistance, Gerhard Glatting for help in dosimetry and Detlev Bartkowiak for providing the gamma-irradiation source.

This work was supported by BMBF IZKF C14 grant and Wilhelm Sander-Stiftung grant 2002.045.1.

References

  1. 1.
    Reske SN, Bunjes D, Buchmann I, Seitz U, Glatting G, Neumaier B, Kotzerke J, Buck A, Martin H, Döhner H, Bergmann L. Targeted bone marrow irradiation in the conditioning of high-risk leukaemia prior to stem cell transplantation. Eur J Nucl Med 2001; 28:807–815.PubMedGoogle Scholar
  2. 2.
    Bunjes D, Buchmann I, Duncker, C, Seitz U, Kotzerke J, Wiesneth M, Dohr D, Stefanic M, Buck A, Harsdorf SV, Glatting G, Grimminger W, Karakas T, Munzert G, Doehner H, Bergmann, L, Reske SN. Rhenium 188-labeled anti-CD66 (a, b, c, e) monoclonal antibody to intensify the conditioning regimen prior to stem cell transplantation for patients with high-risk acute myeloid leukemia or myelodysplastic syndrome: results of a phase I-II study. Blood 2001; 98:565–572.PubMedGoogle Scholar
  3. 3.
    Matthews DC, Appelbaum FR, Eary JF, et al. Radiolabeled anti-CD45 monoclonal antibodies target lymphohematopoietic tissue in the macaque. Blood 1991; 78:1864–1874.PubMedGoogle Scholar
  4. 4.
    Matthews DC, Appelbaum FR, Eary JF, et-al. Development of a marrow transplant regimen for acute leukemia using targeted hematopoietic irradiation delivered by (131)I-labeled anti-CD45 antibody, combined with cyclophosphamide and total body irradiation. Blood 1995; 85:1122–1131.PubMedGoogle Scholar
  5. 5.
    Matthews DC, Appelbaum FR, Eary JF, Fisher DR, Durack LD, Hui TE, Martin PJ, Mitchell D, Press OW, Storb R, Bernstein ID. Phase I study of (131)I-anti-CD45 antibody plus cyclophosphamide and total body irradiation for advanced acute leukemia and myelodysplastic syndrome. Blood 1999; 94:1237–1247.PubMedGoogle Scholar
  6. 6.
    Press OW, Shan D, Howell-Clark J, Eary J, Appelbaum FR, Matthews D, King DJ, Haines AM, Hamann P, Hinman L, Shochat D, Bernstein ID. Comparative metabolism and retention of iodine-125, yttrium-90, and indium-111 radioimmunoconjugates by cancer cells. Cancer Res 1996; 56:2123–2129.PubMedGoogle Scholar
  7. 7.
    Friesen C, Herr I, Krammer PH, Debatin KM. Involvement of the CD95 (APO-1/FAS) receptor/ligand system in drug-induced apoptosis in leukemia cells. Nat Med 1996; 2:574-577.PubMedGoogle Scholar
  8. 8.
    Friesen C, Fulda S, Debatin KM. Drugs and the CD95 pathway. Leukemia 1999; 13:1854–1858.CrossRefPubMedGoogle Scholar
  9. 9.
    Friesen C, Fulda S, Debatin KM. Activation of the CD95 system by doxorubicin is modulated by the redox state in chemosensitive- and drug-resistant tumor cells. Cell Death Differ 1999; 6:471–480.CrossRefPubMedGoogle Scholar
  10. 10.
    Fulda S, Scaffidi C, Pietsch T, Krammer PH, Peter ME, Debatin KM. Activation of the CD95 (APO-1/Fas) pathway in drug- and gamma-irradiation-induced apoptosis of brain tumor cells. Cell Death Differ 1998; 5:884–893.CrossRefPubMedGoogle Scholar
  11. 11.
    Müller M, Strand S, Hug H, Heinemann M, Walczak H, Hofmann WJ, Stremmel W, Krammer PH, Galle PR. Drug-induced apoptosis in hepatoma cells is mediated by the CD95 (APO-1/Fas) receptor/ligand system and involves activation of wild-type p53. J Clin Invest 1997; 99:403–413.PubMedGoogle Scholar
  12. 12.
    Kasibatla S, Brunner T, Genestier L, Echeverri F, Mahboubi A, Green DR. DNA damaging agents induce expression of FAS ligand and subsequent apoptosis in T lymphocytes via activation of NF-κ-B. Mol Cell 1998; 1:543–551.PubMedGoogle Scholar
  13. 13.
    Stahnke K, Fulda S, Friesen C, Strauss G, Debatin KM. Activation of apoptosis pathways in peripheral blood lymphocytes by in vivo chemotherapy. Blood 2001; 98:3066–3073.CrossRefPubMedGoogle Scholar
  14. 14.
    Kaufmann SH, Earnshaw WC. Induction of apoptosis by cancer therapy. Exp Cell Res 2000; 256:42–49.CrossRefPubMedGoogle Scholar
  15. 15.
    Scaffidi C, Fulda S, Li F, Friesen C, Srinivasan A, Tomaselli KJ, Debatin KM, Krammer PH, Peter ME. Two CD95 signaling pathways. EMBO 1998; 17:1675–1687.PubMedGoogle Scholar
  16. 16.
    Kischkel FC, Hellbardt S, Behrmann I, Gremer M, Pawlita M, Krammer PH, Peter M. Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins (CAP) form a death-inducing signalling complex (DISC) with the receptor. EMBO J 1995; 14:5579–5588.PubMedGoogle Scholar
  17. 17.
    Peter M E, Kischkel FC, Hellbrandt S, Chinnaiyan AM, Krammer PH, Dixit VM. CD95 (APO-1/Fas)-associating signaling proteins. Cell Death Differ 1996; 3:161–170.Google Scholar
  18. 18.
    Debatin KM. Disturbances of the CD95 (APO-1/Fas) system in disorders of lymphohaematopoietic cells. Cell Death Differ 1996; 3:185–189.Google Scholar
  19. 19.
    Dhein J, Walczak H, Bäumler C, Debatin KM, Krammer PH. Autocrine T cell suicide mediated by APO-1 (Fas/CD95). Nature 1995; 373:438–441.PubMedGoogle Scholar
  20. 20.
    Rueffli AA, Smyth MJ, Johnstone RW. HMBA induces activation of caspase-independent cell death pathway to overcome P-glycoprotein-mediated multidrug resistance. Blood 2000; 95:2378–2385.PubMedGoogle Scholar
  21. 21.
    Fulda S, Meyer E, Friesen C, Susin SA, Kroemer E, Debatin KM. Cell type specific involvement of death receptor and mitchondrial pathways in drug-induced apoptosis. Oncogene 2001; 20:1063–1075.Google Scholar
  22. 22.
    Eischen CM, Kottke TJ, Martins LM, Basi GS, Tung JS, Earnshaw WC, Leibson PJ, Kaufmann SH. Comparison of apoptosis in wild-type and Fas-resistant cells: chemotherapy-induced apoptosis is not dependent on Fas/Fas ligand interactions. Blood 1997; 90:935–943.PubMedGoogle Scholar
  23. 23.
    Villunger A, Egle A, Kos M, Hartmann BL, Geley S, Kofler R, Greil R. Drug-induced apoptosis is associated with enhanced Fas (Apo-1/CD95) ligand expression but occurs independently of Fas (Apo-1/CD95) signaling in human T-acute lymphatic leukemia cells. Cancer Res 1997; 57:3331–3334.PubMedGoogle Scholar
  24. 24.
    Wesselborg S, Engels IH, Rossmann E, Los M, Schulze-Osthoff K. Anticancer drugs induce caspase-8/FLICE activation and apoptosis in the absence of CD95 receptor/ligand interaction. Blood 1999; 93:3053–3063.PubMedGoogle Scholar
  25. 25.
    McGahon AJ, Pereira Costa AP, Daly L, Cotter TG. Chemotherapeutic drug-induced apoptosis in human leukaemic cells is independent of Fas (APO-1/CD95) receptor/ligand system. Br J Haematol 1998; 101:539–547.CrossRefPubMedGoogle Scholar
  26. 26.
    Li P, Nijhawan D, Budihardjo I, Srinivasula S, Ahmad M, Alnemri AS, Wang X. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997; 91:479–489.PubMedGoogle Scholar
  27. 27.
    Zou H, Henzel WJ, Liu,X, Lutschg A, Wang X. Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell 1997; 90:405–413.PubMedGoogle Scholar
  28. 28.
    Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE, Brothers GM, Mangion J, Jacotot E, Costantini P, Loeffler M, Larochette N, Goodlett DR, Aebersold R, Siderovski DP, Penninger JM, Kroemer G. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 1999; 397:441–446.PubMedGoogle Scholar
  29. 29.
    Susin SA, Lorenzo H., Zamzami N, Marzo I, Brenner C, Larochette N, Prevost MC, Alzari PM, Kroemer G. Mitochondrial release of caspase-2 and caspase-9 during the apoptotic process. J Exp Med 1999; 189:381–394.PubMedGoogle Scholar
  30. 30.
    Srinivasula SM, Hedge R, Saleh A, Datta P, Shiozaki E, Chai J, Lee RA, Robbins PD, Fernandes-Alnemri T, Shi Y, Alnemri E. A conserved XIAP-interaction in caspase-9 and Smac/DIABLO regulates caspase activity and apoptosis. Nature 2001; 410:112–116.CrossRefPubMedGoogle Scholar
  31. 31.
    Deveraux QL, Leo E, Stennicke HR, Welsh K, Salvesen GS, Reed J. Cleavage of human inhibitors of apoptosis protein XIAP results in fragments with distinct specificities for caspases. EMBO J 1999; 18:5242–5251.PubMedGoogle Scholar
  32. 32.
    Kroemer G. The proto-oncogene Bcl-2 and its role in regulating apoptosis. Nat Med 1997; 3:614–620.PubMedGoogle Scholar
  33. 33.
    Hu Y, Benedict MA, Wu D, Inohara N, Nunez G. Bcl-XL interacts with Apaf-1 and inhibits Apaf-1-dependent caspase-9 activation. Proc Natl Acad Sci U S A 1998; 95:4386–4391.PubMedGoogle Scholar
  34. 34.
    Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 1997; 275:1129–1132.PubMedGoogle Scholar
  35. 35.
    Lakin ND, Jackson SP. Regulation of p53 in response to DNA damage. Oncogene 1999; 18:7644–7655.PubMedGoogle Scholar
  36. 36.
    Miyashita T, Reed J. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 1995; 80:293–299.PubMedGoogle Scholar
  37. 37.
    Milner J. DNA damage, p53 and anticancer therapies. Nat Med 1995; 1:879–880.PubMedGoogle Scholar
  38. 38.
    Nicoletti I, Migliorati G, Pagliacci MC, Grignani F,Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 1991; 139:271–279.PubMedGoogle Scholar
  39. 39.
    Carbonari M, Cibati M, Cherchi M, Sbarigia D, Pesce AM, Dell'Ann L, Modica A, Fiorelli M. Detection and characterization of apoptotic peripheral blood lymphocytes in human immunodeficiency virus-infection and cancer chemotherapy by a novel flow immunocytometric method. Blood 1994; 83:1268–1277.PubMedGoogle Scholar
  40. 40.
    Weller M, Winter S, Schmidt C, Esser P, Fontana A, Dichgans J, Groscurth P. Topoisomerase I inhibitors for human malignant glioma: differential modulation of p53, p21, bax and Bcl-2 expression and of CD95-mediated apoptosis by camptothecin and beta lapachone. Int J Cancer 1997; 73:707–714.CrossRefPubMedGoogle Scholar
  41. 41.
    Narita M, Shimizu S, Ito T, Chittenden T, Lutz RJ, Matsuda H, Tsujimoto Y. Bax interacts with the permeability transition pore to induce permeability transition and cytochrome c release in isolated mitochondria. Proc Natl Acad Sci U S A 1998; 95:14681–14686.PubMedGoogle Scholar
  42. 42.
    Shimizu S, Narita M, Tsujimoto Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 1999; 399:483–487.Google Scholar
  43. 43.
    Vogt Sionov R, Haupt Y. The cellular response tp p53: the decision between life and death. Oncogene 1999; 18:6145–6157.CrossRefPubMedGoogle Scholar
  44. 44.
    Ishikawa J, Akimaru K, Nakanishi M, Tomokiyo K, Furuta K, Suzuki M, Noyori R. Anti-cancer-prostaglandin-induced cell-cycle arrest and its modulation by an inhibitor of the ATP-dependent glutathione S-conjugate export pump (GS-X pump). Biochem J 1998; 336:569–576.PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Claudia Friesen
    • 1
  • Annelie Lubatschofski
    • 1
  • Jörg Kotzerke
    • 2
  • Inga Buchmann
    • 2
  • Sven N. Reske
    • 2
  • Klaus-Michael Debatin
    • 1
    Email author
  1. 1.University Children's HospitalUlmGermany
  2. 2.Department of Nuclear MedicineUniversity of UlmUlmGermany

Personalised recommendations