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Sensitization to TRAIL-induced apoptosis and modulation of FLICE-inhibitory protein in B chronic lymphocytic leukemia by actinomycin D

Leukemia volume 15, pages 1868–1877 (2001)Cite this article

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent activator of the cell death pathway and exerts tumoricidal activity in vivo with minimal toxicity. In order to investigate the therapeutic potential of TRAIL in B chronic lymphocytic leukemia (B-CLL) we have analyzed the expression of TRAIL receptors (TRAIL-Rs) in leukemic cells from B-CLL patients and their in vitro sensitivity to apoptosis induced by recombinant human TRAIL. We have found TRAIL-R1 and -R2 death receptor, and TRAIL-R3 and -R4 decoy receptor mRNA expression in most of the 57 B-CLL patients studied (R1 82%, R2 100%, R3 96% and R4 82%). TRAIL-R1 and R2 proteins were expressed on the surface and within the cells, whereas R3 and R4 decoy receptors were almost exclusively expressed in the cytoplasm. Despite TRAIL death receptor expression, B-CLL cells were relatively resistant to induction of apoptosis by recombinant human TRAIL (300 ng/ml). However, the susceptibility to TRAIL-induced apoptosis was increased by treatment of B-CLL cells with actinomycin D (Act D). Western blot analysis showed higher constitutive expression of the long form of FLICE-inhibitory protein (FLIPL) in B-CLL as compared to normal tonsillar B cells. Act D treatment down-regulated both long and short FLIP expression, which was correlated with the increase in B-CLL sensitivity to TRAIL. Although the surface TRAIL death receptor expression was up-regulated both by cell culture and by Act D treatment, the changes were not correlated with a gain in susceptibility to TRAIL. In addition, neither decoy receptors nor Bcl-2 expression were affected by Act D. Our findings suggest the possible involvement of FLIP in regulating TRAIL-mediated apoptosis in B-CLL.

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References

  1. Daniel PT, Wieder T, Sturm I, Schulze-Osthoff K . The kiss of death: promises and failures of death receptors and ligands in cancer therapy Leukemia 2001 15: 1022–1032

    Article  CAS  PubMed  Google Scholar 

  2. Pan G, Ni J, Wei Y-F, Yu G-L, Gentz R, Dixit VM . An antagonist decoy receptor and a death domain-containing receptor for TRAIL Science 1997 277: 815–818

    Article  CAS  PubMed  Google Scholar 

  3. Pan G, O'Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, Dixit VM . The receptor for the cytotoxic ligand TRAIL Science 1997 276: 111–113

    Article  CAS  PubMed  Google Scholar 

  4. Walczak H, Degli-Esposti MA, Johnson RS, Smolak PJ, Waugh JY, Boiani N, Timour MS, Gerhart MJ, Schooley KA, Smith CA, Goodwin RG, Rauch CT . TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL EMBO J 1997 16: 5386–5397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A . Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors Science 1997 277: 818–821

    Article  CAS  PubMed  Google Scholar 

  6. Degli-Esposti MA, Smolak PJ, Walczak H, Waugh J, Huang C-P, DuBose RF, Goodwin RG, Smith CA . Cloning and characterization of TRAIL-R3, a novel member of the emerging TRAIL receptor family J Exp Med 1997 186: 1165–1170

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. MacFarlane M, Ahmad M, Srinivasula SM, Fernandes-Alnemri T, Cohen GM, Alnemri ES . Identification and molecular cloning of two novel receptors for the cytotoxic ligand TRAIL J Biol Chem 1997 272: 25417–25420

    Article  CAS  PubMed  Google Scholar 

  8. Degli-Esposti MA, Dougall WC, Smolak PJ, Waugh JY, Smith CA, Goodwin RG . The novel receptor TRAIL-R4 induces NF-κB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain Immunity 1997 7: 813–820

    Article  CAS  PubMed  Google Scholar 

  9. Marsters SA, Sheridan JP, Pitti RM, Huang A, Skubatch M, Baldwin D, Yuan J, Gurney A, Goddard AD, Godowski P, Ashkenazi A . A novel receptor for Apo2L/TRAIL contains a truncated death domain Curr Biol 1997 7: 1003–1006

    Article  CAS  PubMed  Google Scholar 

  10. Griffith TS, Lynch DH . TRAIL: a molecule with multiple receptors and control mechanisms Curr Opin Immunol 1998 10: 559–563

    Article  CAS  PubMed  Google Scholar 

  11. Griffith TS, Chin WA, Jackson GC, Lynch DH, Kubin MZ . Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells J Immunol 1998 161: 2833–2840

    CAS  PubMed  Google Scholar 

  12. Walczak H, Miller RE, Ariail K, Gliniak B, Griffith TS, Kubin M, Chin W, Jones J, Woodward A, Le T, Smith C, Smolak P, Goodwin RG, Rauch CT, Schuh JCL, Lynch DH . Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo Nat Med 1999 5: 157–163

    Article  CAS  PubMed  Google Scholar 

  13. Griffith TS, Rauch CT, Smolak PJ, Waugh JY, Boiani N, Lynch DH, Smith CA, Goodwin RG, Kubin MZ . Functional analysis of TRAIL receptors using monoclonal antibodies J Immunol 1999 162: 2597–2605

    CAS  PubMed  Google Scholar 

  14. Thome M, Schneider P, Hofmann K, Fickenscher H, Meinl E, Neipel F, Mattmann C, Burns K, Bodmer J-L, Schroter M, Scaffidi C, Krammer PH, Peter ME, Tschopp J . Viral FLICE-inhibitory proteins (FLIPs) prevent apoptosis induced by death receptors Nature 1997 386: 517–521

    Article  CAS  PubMed  Google Scholar 

  15. Irmler M, Thome M, Hahne M, Schneider P, Hofmann K, Steiner V, Bodmer J-L Schröter M, Burns K, Mattmann C, Rimoldi D, French LE, Tschopp J . Inhibition of death receptor signals by cellular FLIP Nature 1997 388: 190–195

    Article  CAS  PubMed  Google Scholar 

  16. Krueger A, Schmitz I, Baumann S, Krammer PH, Kirchhoff S . Cellular FLICE-inhibitory protein splice variants inhibit different steps of caspase-8 activation at the CD95 death-inducing signaling complex J Biol Chem 2001 276: 20633–20640

    Article  CAS  PubMed  Google Scholar 

  17. Scaffidi C, Schmitz I, Krammer PH, Peter ME . The role of c-FLIP in modulation of CD95-induced apoptosis J Biol Chem 1999 274: 1541–1548

    Article  CAS  PubMed  Google Scholar 

  18. Medema JP, de Jong J, van Hall T, Melief CJ, Offringa R . Immune escape of tumors in vivo by expression of cellular FLICE-inhibitory protein J Exp Med 1999 190: 1033–1038

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Djerbi M, Screpanti V, Catrina AI, Bogen B, Biberfeld P, Grandien A . The inhibitor of death receptor signaling, FLICE-inhibitory protein defines a new class of tumor progression factors J Exp Med 1999 190: 1025–1032

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Kim K, Fisher MJ, Xu SQ, El-Deiry WS . Molecular determinants of response to TRAIL in killing of normal and cancer cells Clin Cancer Res 2000 6: 335–346

    CAS  PubMed  Google Scholar 

  21. Leverkus M, Neumann M, Mengling T, Rauch CT, Brocker EB, Krammer PH, Walczak H . Regulation of tumor necrosis factor-related apoptosis-inducing ligand sensitivity in primary and transformed human keratinocytes Cancer Res 2000 60: 553–559

    CAS  PubMed  Google Scholar 

  22. Deveraux QL, Reed JC . IAP family proteins – suppressors of apoptosis Genes Dev 1999 13: 239–252

    Article  CAS  PubMed  Google Scholar 

  23. Strasser A, Harris AW, Huang DC, Krammer PH, Cory S . Bcl-2 and Fas/APO-1 regulate distinct pathways to lymphocyte apoptosis EMBO J 1995 14: 6136–6147

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Ashkenazi A, Pai RC, Fong S, Leung S, Lawrence DA, Marsters SA, Blackie C, Chang L, McMurtrey AE, Hebert A, DeForge L, Koumenis IL, Lewis D, Harris L, Bussiere J, Koeppen H, Shahrokh Z, Schwall RH . Safety and antitumor activity of recombinant soluble Apo2 ligand J Clin Invest 1999 104: 155–162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Jurlander J . The cellular biology of B-cell chronic lymphocytic leukemia Crit Rev Oncol Hematol 1998 27: 29–52

    Article  CAS  PubMed  Google Scholar 

  26. O'Brien S, del Giglio A, Keating M . Advances in the biology and treatment of B-cell chronic lymphocytic leukemia Blood 1995 85: 307–318

    CAS  PubMed  Google Scholar 

  27. Binet JL, Plunkett W, Robertson B, Merle-Beral H, Mentz F, Hoffbrand AV, Panayiotidis P . What does apoptosis mean in CLL? Leuk Lymphoma 1996 22: 47–52

    Article  PubMed  Google Scholar 

  28. Wang D, Freeman GJ, Levine H, Ritz J, Robertson MJ . Role of the CD40 and CD95 (APO-1/Fas) antigens in the apoptosis of human B-cell malignancies Br J Haematol 1997 97: 409–417

    Article  CAS  PubMed  Google Scholar 

  29. Tinhofer I, Marschitz I, Kos M, Henn T, Egle A, Villunger A, Greil R . Differential sensitivity of CD4+ and CD8+ T lymphocytes to the killing efficacy of Fas (APO-1/CD95) ligand+ tumor cells in B chronic lymphocytic leukemia Blood 1998 91: 4273–4281

    CAS  PubMed  Google Scholar 

  30. McConkey DJ, Chandra J, Wright S, Plunkett W, McDonnell TJ, Reed JC, Keating M . Apoptosis sensitivity in chronic lymphocytic leukemia is determined by endogenous endonuclease content and relative expression of BCL-2 and BAX J Immunol 1996 156: 2624–2630

    CAS  PubMed  Google Scholar 

  31. Thomas A, El Rouby S, Reed JC, Krajewski S, Silber R, Potmesil M, Newcomb EW . Drug-induced apoptosis in B-cell chronic lymphocytic leukemia: relationship between p53 gene mutation and bcl-2/bax proteins in drug resistance Oncogene 1996 12: 1055–1062

    CAS  PubMed  Google Scholar 

  32. Pepper C, Bentley P, Hoy T . Regulation of clinical chemoresistance by bcl-2 and bax oncoproteins in B-cell chronic lymphocytic leukaemia Br J Haematol 1996 95: 513–517

    Article  CAS  PubMed  Google Scholar 

  33. Aguilar-Santelises M, Rottenberg ME, Lewin N, Mellstedt H, Jondal M . Bcl-2, Bax and p53 expression in B-CLL in relation to in vitro survival and clinical progression Int J Cancer 1996 69: 114–119

    Article  CAS  PubMed  Google Scholar 

  34. Osorio LM, De Santiago A, Aguilar-Santelises M, Mellstedt H, Jondal M . CD6 ligation modulates the Bcl-2/Bax ratio and protects chronic lymphocytic leukemia B cells from apoptosis induced by anti-IgM Blood 1997 89: 2833–2841

    CAS  PubMed  Google Scholar 

  35. Rai KR, Sawitsky A, Cronkite EP, Chanana AD, Levy RN, Pasternack BS . Clinical staging of chronic lymphocytic leukemia Blood 1975 46: 219–234

    CAS  PubMed  Google Scholar 

  36. Silver RT, Sawitsky A, Rai K, Holland JF, Glidewell O . Guidelines for protocol studies in chronic lymphocytic leukemia Am J Hematol 1978 4: 343–358

    Article  CAS  PubMed  Google Scholar 

  37. Zhang XD, Franco A, Myers K, Gray C, Nguyen T, Hersey P . Relation of TNF-related apoptosis-inducing ligand (TRAIL) receptor and FLICE-inhibitory protein expression to TRAIL-induced apoptosis of melanoma Cancer Res 1999 59: 2747–2753

    CAS  PubMed  Google Scholar 

  38. Zamzami N, Marchetti P, Castedo M, Zanin C, Vayssiere J-L, Petit PX, Kroemer G . Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo J Exp Med 1995 181: 1661–1672

    Article  CAS  PubMed  Google Scholar 

  39. Martinez-Lorenzo MJ, Alava MA, Gamen S, Kim KJ, Chuntharapai A, Pineiro A, Naval J, Anel A . Involvement of APO2 ligand/TRAIL in activation-induced death of Jurkat and human peripheral blood T cells Eur J Immunol 1998 28: 2714–2725

    Article  CAS  PubMed  Google Scholar 

  40. Mori S, Murakami-Mori K, Nakamura S, Ashkenazi A, Bonavida B . Sensitization of AIDS-Kaposi's sarcoma cells to Apo-2 ligand-induced apoptosis by actinomycin D J Immunol 1999 162: 5616–5623

    CAS  PubMed  Google Scholar 

  41. Suliman A, Lam A, Datta R, Srivastava RK . Intracellular mechanisms of TRAIL: apoptosis through mitochondrial-dependent and -independent pathways Oncogene 2001 20: 2122–2133

    Article  CAS  PubMed  Google Scholar 

  42. Wen J, Ramadevi N, Nguyen D, Perkins C, Worthington E, Bhalla K . Antileukemic drugs increase death receptor 5 levels and enhance Apo-2L-induced apoptosis of human acute leukemia cells Blood 2000 96: 3900–3906

    CAS  PubMed  Google Scholar 

  43. Zhang XD, Franco AV, Nguyen T, Gray CP, Hersey P . Differential localization and regulation of death and decoy receptors for TNF-related apoptosis-inducing ligand (TRAIL) in human melanoma cells J Immunol 2000 164: 3961–3970

    Article  CAS  PubMed  Google Scholar 

  44. Nguyen T, Zhang XD, Hersey P . Relative resistance of fresh isolates of melanoma to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis Clin Cancer Res 2001 7: 966s–973s

    CAS  PubMed  Google Scholar 

  45. Keane MM, Ettenberg SA, Nau MM, Russell EK, Lipkowitz S . Chemotherapy augments TRAIL-induced apoptosis in breast cell lines Cancer Res 1999 59: 734–741

    CAS  PubMed  Google Scholar 

  46. Frank S, Kohler U, Schackert G, Schackert HK . Expression of TRAIL and its receptors in human brain tumors Biochem Biophys Res Commun 1999 257: 454–459

    Article  CAS  PubMed  Google Scholar 

  47. Degli-Esposti M . To die or not to die – the quest of the TRAIL receptors J Leukoc Biol 1999 65: 535–542

    Article  CAS  PubMed  Google Scholar 

  48. Bonavida B, Ng C-P, Jazirehi A, Schiller G, Mizutani Y . Selectivity of TRAIL-mediated apoptosis of cancer cells and synergy with drugs: the trail to non-toxic cancer therapeutics Int J Oncol 1999 15: 793–802

    CAS  PubMed  Google Scholar 

  49. Kischkel FC, Lawrence DA, Chuntharapai A, Schow P, Kim KJ, Ashkenazi A . Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5 Immunity 2000 12: 611–620

    Article  CAS  PubMed  Google Scholar 

  50. Sprick MR, Weigand MA, Rieser E, Rauch CT, Juo P, Blenis J, Krammer PH, Walczak H . FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2 Immunity 2000 12: 599–609

    Article  CAS  PubMed  Google Scholar 

  51. Abe K, Kurakin A, Mohseni-Maybodi M, Kay B, Khosravi-Far R . The complexity of TNF-related apoptosis-inducing ligand Ann N Y Acad Sci 2000 926: 52–63

    Article  CAS  PubMed  Google Scholar 

  52. Fulda S, Meyer E, Debatin KM . Metabolic inhibitors sensitize for CD95 (APO-1/Fas)-induced apoptosis by down-regulating Fas-associated death domain-like interleukin 1-converting enzyme inhibitory protein expression Cancer Res 2000 60: 3947–3956

    CAS  PubMed  Google Scholar 

  53. Kataoka T, Budd RC, Holler N, Thome M, Martinon F, Irmler M, Burns K, Hahne M, Kennedy N, Kovacsovics M, Tschopp J . The caspase-8 inhibitor FLIP promotes activation of NF-kappaB and Erk signaling pathways Curr Biol 2000 10: 640–648

    Article  CAS  PubMed  Google Scholar 

  54. Kitada S, Zapata JM, Andreeff M, Reed JC . Bryostatin and CD40-ligand enhance apoptosis resistance and induce expression of cell survival genes in B-cell chronic lymphocytic leukemia Br J Haematol 1999 106: 995–1004

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are indebted to Dr Stewart D Lyman and Dr Tony Troutt (Immunex, Seattle, WA, USA) for their generous gift of the TRAIL reagents, and to Prof Alf Grandien (Stockholm University, Sweden) for the FLIP transfected murine B lymphoma A20 cell lines. This work was supported by funds from the Swedish Cancer Association, the Swedish Medical Association and the Karolinska Institutet.

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Authors and Affiliations

  1. Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden

    A Olsson, T Diaz, M Jondal & LM Osorio

  2. Women's Hospital, ENH Research Institute, Evanston, USA

    M Aguilar-Santelises

  3. Department of Hematology and Infectious Diseases, Karolinska Hospital, Stockholm, Sweden

    A Österborg & F Celsing

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Olsson, A., Diaz, T., Aguilar-Santelises, M. et al. Sensitization to TRAIL-induced apoptosis and modulation of FLICE-inhibitory protein in B chronic lymphocytic leukemia by actinomycin D. Leukemia 15, 1868–1877 (2001). https://doi.org/10.1038/sj.leu.2402287

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