Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Manuscript
  • Published:

Apoptose

Induction of apoptosis by proteasome inhibitors in B-CLL cells is associated with downregulation of CD23 and inactivation of Notch2

Leukemia volume 19, pages 260–267 (2005)Cite this article

Abstract

Recently, proteasome inhibitors (PI) have attracted interest as novel anticancer agents in B-cell chronic lymphocytic leukemia (B-CLL). A prominent feature of B-CLL cells is the high expression of CD23, which is closely related to cell survival and is regulated by Notch2. Since several components of the Notch signaling cascade are tightly regulated by proteasomal degradation, we studied the effect of PI on Notch2 activity and CD23 expression. Exposure of B-CLL cells to PI led to induction of apoptosis, a time- and dose-dependent downregulation of CD23 expression and a decline in DNA binding of transcriptionally active Notch2. In contrast, the transcription factor NF-AT and its putative target gene CD5, which is highly expressed in B-CLL cells, were unaffected. When the late phase of PI-induced apoptosis was arrested by inhibition of caspase 3, the reduction of Notch2 activity was still observed, indicating that reduction of active Notch2 took place already during an earlier phase of apoptosis. Enforced expression of constitutively active Notch2 decreased PI-mediated apoptosis in a human B-cell line. These data indicate that downregulation of CD23 and loss of Notch2 activity are early steps in PI-induced apoptosis of B-CLL lymphocytes and may be part of the full apoptotic response.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Kay NE, Hamblin TJ, Jelinek DF, Dewald GW, Byrd JC, Farag S et al. Chronic lymphocytic leukemia. Hematology (Am Soc Hematol Educ Program) 2002; 175: 193–213.

    Article  Google Scholar 

  2. Guipaud O, Deriano L, Salin H, Vallat L, Sabatier L, Merle-Beral H et al. B-cell chronic lymphocytic leukaemia: a polymorphic family unified by genomic features. Lancet Oncol 2003; 4: 505–514.

    Article  Google Scholar 

  3. Jewell AP . Role of apoptosis in the pathogenesis of B-cell chronic lymphocytic leukaemia. Br J Biomed Sci 2002; 59: 235–238.

    Article  Google Scholar 

  4. Fournier S, Yang LP, Delespesse G, Rubio M, Biron G, Sarfati M . The two CD23 isoforms display differential regulation in chronic lymphocytic leukaemia. Br J Haematol 1995; 89: 373–379.

    Article  CAS  Google Scholar 

  5. Hubmann R, Schwarzmeier JD, Shehata M, Hilgarth M, Duechler M, Dettke M et al. Notch2 is involved in the overexpression of CD23 in B-cell chronic lymphocytic leukemia. Blood 2002; 99: 3742–3747.

    Article  CAS  Google Scholar 

  6. Artavanis-Tsakonas S, Rand MD, Lake RJ . Notch signaling: cell fate control and signal integration in development. Science 1999; 284: 770–776.

    Article  CAS  Google Scholar 

  7. Jarriault S, Brou C, Logeat F, Schroeter EH, Kopan R, Israel A . Signalling downstream of activated mammalian Notch. Nature 1995; 377: 355–358.

    Article  CAS  Google Scholar 

  8. Schroeter EH, Kisslinger JA, Kopan R . Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain. Nature 1998; 393: 382–386.

    Article  CAS  Google Scholar 

  9. Berezovska O, Jack C, McLean P, Aster JC, Hicks C, Xia W et al. Rapid Notch1 nuclear translocation after ligand binding depends on presenilin-associated gamma-secretase activity. Ann N Y Acad Sci 2000; 920: 223–226.

    Article  CAS  Google Scholar 

  10. Ellisen LW, Bird J, West DC, Soreng AL, Reynolds TC, Smith SD et al. TAN-1, the human homolog of the Drosophila notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms. Cell 1991; 66: 649–661.

    Article  CAS  Google Scholar 

  11. Capobianco AJ, Zagouras P, Blaumueller CM, Artavanis-Tsakonas S, Bishop JM . Neoplastic transformation by truncated alleles of human NOTCH1/TAN1 and NOTCH2. Mol Cell Biol 1997; 17: 6265–6273.

    Article  CAS  Google Scholar 

  12. Witt CM, Hurez V, Swindle CS, Hamada Y, Klug CA . Activated Notch2 potentiates CD8 lineage maturation and promotes the selective development of B1 B cells. Mol Cell Biol 2003; 23: 8637–8650.

    Article  CAS  Google Scholar 

  13. Bertrand FE, Eckfeldt CE, Lysholm AS, LeBien TW . Notch-1 and Notch-2 exhibit unique patterns of expression in human B-lineage cells. Leukemia 2000; 14: 2095–2102.

    Article  CAS  Google Scholar 

  14. Witt CM, Won WJ, Hurez V, Klug CA . Notch2 haploinsufficiency results in diminished B1 B cells and a severe reduction in marginal zone B cells. J Immunol 2003; 171: 2783–2788.

    Article  CAS  Google Scholar 

  15. Saito T, Chiba S, Ichikawa M, Kunisato A, Asai T, Shimizu K et al. Notch2 is preferentially expressed in mature B cells and indispensable for marginal zone B lineage development. Immunity 2003; 18: 675–685.

    Article  CAS  Google Scholar 

  16. Deftos ML, He YW, Ojala EW, Bevan MJ . Correlating notch signaling with thymocyte maturation. Immunity 1998; 9: 777–786.

    Article  CAS  Google Scholar 

  17. Jehn BM, Bielke W, Pear WS, Osborne BA . Cutting edge: protective effects of notch-1 on TCR-induced apoptosis. J Immunol 1999; 162: 635–638.

    CAS  PubMed  Google Scholar 

  18. Miele L, Osborne B . Arbiter of differentiation and death: notch signaling meets apoptosis. J Cell Physiol 1999; 181: 393–409.

    Article  CAS  Google Scholar 

  19. Shelly LL, Fuchs C, Miele L . Notch-1 inhibits apoptosis in murine erythroleukemia cells and is necessary for differentiation induced by hybrid polar compounds. J Cell Biochem 1999; 73: 164–175.

    Article  CAS  Google Scholar 

  20. Sade H, Krishna S, Sarin A . The anti-apoptotic effect of Notch-1 requires p56lck-dependent, Akt/PKB-mediated signaling in T cells. J Biol Chem 2004; 279: 2937–2944.

    Article  CAS  Google Scholar 

  21. Qiu L, Joazeiro C, Fang N, Wang HY, Elly C, Altman Y et al. Recognition and ubiquitination of Notch by Itch, a hect-type E3 ubiquitin ligase. J Biol Chem 2000; 275: 35734–35737.

    Article  CAS  Google Scholar 

  22. Wu G, Lyapina S, Das I, Li J, Gurney M, Pauley A et al. SEL-10 is an inhibitor of notch signaling that targets notch for ubiquitin-mediated protein degradation. Mol Cell Biol 2001; 21: 7403–7415.

    Article  CAS  Google Scholar 

  23. Oberg C, Li J, Pauley A, Wolf E, Gurney M, Lendahl U . The Notch intracellular domain is ubiquitinated and negatively regulated by the mammalian Sel-10 homolog. J Biol Chem 2001; 276: 35847–35853.

    Article  CAS  Google Scholar 

  24. Jehn BM, Dittert I, Beyer S, von der MK, Bielke W . c-Cbl binding and ubiquitin-dependent lysosomal degradation of membrane-associated Notch1. J Biol Chem 2002; 277: 8033–8040.

    Article  CAS  Google Scholar 

  25. Naujokat C, Hoffmann S . Role and function of the 26S proteasome in proliferation and apoptosis. Lab Invest 2002; 82: 965–980.

    Article  CAS  Google Scholar 

  26. Ciechanover A . The ubiquitin proteolytic system and pathogenesis of human diseases: a novel platform for mechanism-based drug targeting. Biochem Soc Trans 2003; 31: 474–481.

    Article  CAS  Google Scholar 

  27. Adams J . Proteasome inhibitors as new anticancer drugs. Curr Opin Oncol 2002; 14: 628–634.

    Article  CAS  Google Scholar 

  28. Gillessen S, Groettup M, Cerny T . The proteasome, a new target for cancer therapy. Onkologie 2002; 25: 534–539.

    CAS  PubMed  Google Scholar 

  29. Masdehors P, Merle-Beral H, Magdelenat H, Delic J . Ubiquitin–proteasome system and increased sensitivity of B-CLL lymphocytes to apoptotic death activation. Leuk Lymphoma 2000; 38: 499–504.

    Article  CAS  Google Scholar 

  30. Schenkein D . Proteasome inhibitors in the treatment of B-cell malignancies. Clin Lymphoma 2002; 3: 49–55.

    Article  CAS  Google Scholar 

  31. Pahler JC, Ruiz S, Niemer I, Calvert LR, Andreeff M, Keating M et al. Effects of the proteasome inhibitor, bortezomib, on apoptosis in isolated lymphocytes obtained from patients with chronic lymphocytic leukemia. Clin Cancer Res 2003; 9: 4570–4577.

    CAS  PubMed  Google Scholar 

  32. Wong SC, Chew WK, Tan JE, Melendez AJ, Francis F, Lam KP . Peritoneal CD5+ B-1 cells have signaling properties similar to tolerant B cells. J Biol Chem 2002; 277: 30707–30715.

    Article  CAS  Google Scholar 

  33. Lenz HJ . Clinical update: proteasome inhibitors in solid tumors. Cancer Treat Rev 2003; 29 (Suppl 1): 41–48.

    Article  CAS  Google Scholar 

  34. Magill L, Walker B, Irvine AE . The proteasome: a novel therapeutic target in haematopoietic malignancy. Hematology 2003; 8: 275–283.

    Article  CAS  Google Scholar 

  35. Masdehors P, Omura S, Merle-Beral H, Mentz F, Cosset JM, Dumont J et al. Increased sensitivity of CLL-derived lymphocytes to apoptotic death activation by the proteasome-specific inhibitor lactacystin. Br J Haematol 1999; 105: 752–757.

    Article  CAS  Google Scholar 

  36. Ling PD, Hsieh JJ, Ruf IK, Rawlins DR, Hayward SD . EBNA-2 upregulation of Epstein–Barr virus latency promoters and the cellular CD23 promoter utilizes a common targeting intermediate, CBF1. J Virol 1994; 68: 5375–5383.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Mumm JS, Schroeter EH, Saxena MT, Griesemer A, Tian X, Pan DJ et al. A ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1. Mol Cell 2000; 5: 197–206.

    Article  CAS  Google Scholar 

  38. Das I, Craig C, Funahashi Y, Jung KM, Kim TW, Byers R et al. Notch oncoproteins depend on gamma-secretase/prencilin activity for processing and function. J Biol Chem 2004; 279: 30771–30780.

    Article  CAS  Google Scholar 

  39. Schuh K, Avots A, Tony HP, Serfling E, Kneitz C . Nuclear NF-ATp is a hallmark of unstimulated B cells from B-CLL patients. Leuk Lymphoma 1996; 23: 583–592.

    Article  CAS  Google Scholar 

  40. Berland R, Wortis HH . An NFAT-dependent enhancer is necessary for anti-IgM-mediated induction of murine CD5 expression in primary splenic B cells. J Immunol 1998; 161: 277–285.

    CAS  PubMed  Google Scholar 

  41. Adams J . Preclinical and clinical evaluation of proteasome inhibitor PS-341 for the treatment of cancer. Curr Opin Chem Biol 2002; 6: 493–500.

    Article  CAS  Google Scholar 

  42. Dewson G, Snowden RT, Almond JB, Dyer MJ, Cohen GM . Conformational change and mitochondrial translocation of Bax accompany proteasome inhibitor-induced apoptosis of chronic lymphocytic leukemic cells. Oncogene 2003; 22: 2643–2654.

    Article  CAS  Google Scholar 

  43. Almond JB, Snowden RT, Hunter A, Dinsdale D, Cain K, Cohen GM . Proteasome inhibitor-induced apoptosis of B-chronic lymphocytic leukaemia cells involves cytochrome c release and caspase activation, accompanied by formation of an approximately 700 kDa Apaf-1 containing apoptosome complex. Leukemia 2001; 15: 1388–1397.

    Article  CAS  Google Scholar 

  44. Jesenberger V, Jentsch S . Deadly encounter: ubiquitin meets apoptosis. Nat Rev Mol Cell Biol 2002; 3: 112–121.

    Article  CAS  Google Scholar 

  45. Masdehors P, Merle-Beral H, Maloum K, Omura S, Magdelenat H, Delic J . Deregulation of the ubiquitin system and p53 proteolysis modify the apoptotic response in B-CLL lymphocytes. Blood 2000; 96: 269–274.

    CAS  PubMed  Google Scholar 

  46. Rangarajan A, Syal R, Selvarajah S, Chakrabarti O, Sarin A, Krishna S . Activated Notch1 signaling cooperates with papillomavirus oncogenes in transformation and generates resistance to apoptosis on matrix withdrawal through PKB/Akt. Virology 2001; 286: 23–30.

    Article  CAS  Google Scholar 

  47. Zlobin A, Jang M, Miele L . Toward the rational design of cell fate modifiers: notch signaling as a target for novel biopharmaceuticals. Curr Pharm Biotechnol 2000; 1: 83–106.

    Article  CAS  Google Scholar 

  48. Aghajanian C, Soignet S, Dizon DS, Pien CS, Adams J, Elliott PJ et al. A phase I trial of the novel proteasome inhibitor PS341 in advanced solid tumor malignancies. Clin Cancer Res 2002; 8: 2505–2511.

    CAS  PubMed  Google Scholar 

  49. Adams J . Potential for proteasome inhibition in the treatment of cancer. Drug Discov Today 2003; 8: 307–315.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Grant No. P15100 from the Austrian Science Foundation, from the Austrian National Bank, ‘Jubilaeumsfonds’-project No. 9448, and from the ‘Fellinger Krebsforschungsfonds’.

Author information

Author notes

  1. M Duechler and M Shehata: These two authors contributed equally to this work.

Authors and Affiliations

  1. Ludwig Boltzmann Institute for Cytokine Research, Medical University Vienna, Vienna, Austria

    M Duechler, M Shehata, J D Schwarzmeier, A Hoelbl & R Hubmann

  2. Clinic of Internal Medicine I, Department of Hematology, Medical University Vienna, Waehringer Guertel, Vienna, Austria

    M Shehata, J D Schwarzmeier & M Hilgarth

Authors
  1. M Duechler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M Shehata
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J D Schwarzmeier
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Hoelbl
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M Hilgarth
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R Hubmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J D Schwarzmeier.

Additional information

Supplementary Information

Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu).

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Duechler, M., Shehata, M., Schwarzmeier, J. et al. Induction of apoptosis by proteasome inhibitors in B-CLL cells is associated with downregulation of CD23 and inactivation of Notch2. Leukemia 19, 260–267 (2005). https://doi.org/10.1038/sj.leu.2403592

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2403592

Keywords

This article is cited by

Search

Advanced search

Quick links