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Chronic Lymphocytic Leukemia

ZAP-70 expression, as detected by immunohistochemistry on bone marrow biopsies from early-phase CLL patients, is a strong adverse prognostic factor

Leukemia volume 21, pages 102–109 (2007)Cite this article

Abstract

Zeta-associated protein-70 (ZAP-70), mostly assessed by flow-cytometry (FC), recently emerged as reliable prognostic factor in chronic lymphocytic leukaemia (CLL) at presentation. We evaluated ZAP-70 expression in 156 CLL patients by immunohistochemistry (IHC) on formalin-fixed bone marrow (BM) biopsies at diagnosis. At presentation, 117 patients (75%) were with Binet stage A, 27 (17%) stage B and 12 (8%) stage C. Median follow-up was 61 months (range 6–242). ZAP-70 was expressed in neoplastic lymphocytes of 69 patients (44%). Concordance between ZAP-70 by IHC and ZAP-70 by FC, immunoglobulin heavy chain variable genes (IGHV) mutational status and CD38 expression was found in 41/46 (89%), 41/49 (80%) and in 60/88 (68%) tested cases, respectively. ZAP-70 expression significantly correlated with advanced Binet stage (B–C), diffuse BM infiltration, increased lactate dehydrogenase (LDH) and β2-microglobulin serum levels and lymphocyte doubling time <12 months. ZAP-70 positivity was significantly related to poorer time to progression (median 16 months vs 158 of ZAP-70-negative cases) (P<0.0001) and overall survival (median 106 months vs not reached) (P=0.0002); this correlation was confirmed at multivariate analysis. ZAP-70 expression correlated with poorer outcome also when evaluated only in the 117 stage A patients. In conclusion, immunohistological detection of ZAP-70 on formalin-fixed BM biopsies at diagnosis appears a useful methodological approach to identify patients with poor prognosis in CLL.

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References

  1. Oscier DG, Gardiner AC, Mould SJ, Gilde S, Davis ZA, Ibbotson RE et al. Multivariate analysis of prognostic factors in CLL: clinical stage, IGVH gene mutational status, and loss or mutation of the p53 gene are independent prognostic factors. Blood 2002; 100: 1177–1184.

    CAS  PubMed  Google Scholar 

  2. Fais F, Ghiotto F, Hashimoto S, Sellars B, Valetto A, Allen SL et al. Chronic lymphocytic leukemia B cells express restricted sets of mutated and unmutated antigen receptors. J Clin Invest 1998; 102: 1515–1525.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Hamblin TJ, Davis Z, Gardiner A, Oscier DG, Stevenson FK . Unmutated Ig V(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood 1999; 94: 1848–1854.

    CAS  PubMed  Google Scholar 

  4. Maloum K, Davi F, Merle-Beral H, Pritsch O, Magnac C, Vuillier F et al. Expression of unmutated VH genes is a detrimental prognostic factor in chronic lymphocytic leukemia. Blood 2000; 96: 377–379.

    CAS  PubMed  Google Scholar 

  5. Klein U, Tu Y, Stolovitzky GA, Mattioli M, Cattoretti G, Husson H et al. Gene expression profiling of B cell chronic lymphocytic leukemia reveals a homogeneous phenotype related to memory B cells. J Exp Med 2001; 194: 1625–1638.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Rosenwald A, Alizadeh AA, Widhopf G, Simon R, Davis RE, Yu X et al. Relation of gene expression phenotype to immunoglobulin mutation genotype in B cell chronic lymphocytic leukemia. J Exp Med 2001; 194: 1639–1647.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Chan AC, van Oers NS, Tran A, Turka L, Law CL, Ryan JC et al. Differential expression of ZAP-70 and Syk protein tyrosine kinases, and the role of this family of protein tyrosine kinases in TCR signaling. J Immunol 1994; 152: 4758–4766.

    CAS  PubMed  Google Scholar 

  8. Chen L, Apgar J, Huynh L, Dicker F, Giago-McGahan T, Rassenti L et al. ZAP-70 directly enhances IgM signaling in chronic lymphocytic leukemia. Blood 2005; 105: 2036–2041.

    Article  CAS  PubMed  Google Scholar 

  9. Richardson SJ, Matthews C, Catherwood MA, Alexander HD, Carey BS, Farrugia J et al. ZAP-70 expression is associated with enhanced ability to respond to migratory and survival signals in B cell chronic lymphocytic leukaemia (B-CLL). Blood 2005; 107: 3584–3592.

    Article  PubMed  Google Scholar 

  10. Wiestner A, Rosenwald A, Barry TS, Wright G, Davis ED, Henrickson SE et al. ZAP-70 expression identifies a chronic lymphocytic leukemia subtype with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. Blood 2003; 101: 4944–4951.

    Article  CAS  PubMed  Google Scholar 

  11. Durig J, Nuckel H, Cremer M, Fuhrer A, Halfmeyer K, Fandrey J et al. ZAP-70 expression is a prognostic factor in chronic lymphocytic leukemia. Leukemia 2003; 17: 2426–2434.

    Article  CAS  PubMed  Google Scholar 

  12. Crespo M, Bosch F, Villamor N, Bellosillo B, Colomer D, Rozman M et al. ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia. N Engl J Med 2003; 348: 1764–1775.

    Article  CAS  PubMed  Google Scholar 

  13. Orchard JA, Ibbotson RE, Davis Z, Wiestner A, Rosenwald A, Thomas PW et al. ZAP-70 expression and prognosis in chronic lymphocytic leukaemia. Lancet 2004; 363: 105–111.

    Article  CAS  PubMed  Google Scholar 

  14. Rassenti LZ, Huynh L, Toy TL, Chen L, Keating MJ, Gribben JG et al. ZAP-70 compared with immunoglobulin heavy-chain gene mutation status as a predictor of disease progression in chronic lymphocytic leukemia. N Engl J Med 2004; 351: 893–901.

    Article  CAS  PubMed  Google Scholar 

  15. Del Giudice I, Morilla A, Osuji N, Matutes E, Morilla R, Burford A et al. Zeta-chain associated protein 70 and CD38 combined predict the time to first treatment in patients with chronic lymphocytic leukemia. Cancer 2005; 104: 2124–2132.

    Article  CAS  PubMed  Google Scholar 

  16. Admirand JH, Rassidakis GZ, Abruzzo LV, Valbuena JR, Jones D, Medeiros LJ . Immunohistochemical detection of ZAP-70 in 341 cases of non-Hodgkin and Hodgkin lymphoma. Mod Pathol 2004; 17: 954–961.

    Article  CAS  PubMed  Google Scholar 

  17. Carreras J, Villamor N, Colomo L, Moreno C, Ramon y Cajal S, Crespo M et al. Immunohistochemical analysis of ZAP-70 expression in B-cell lymphoid neoplasms. J Pathol 2005; 205: 507–513.

    Article  CAS  PubMed  Google Scholar 

  18. Rozman C, Montserrat E, Rodriguez-Fernandez JM, Ayats R, Vallespi T, Parody R et al. Bone marrow histologic pattern – the best single prognostic parameter in chronic lymphocytic leukemia: a multivariate survival analysis of 329 cases. Blood 1984; 64: 642–648.

    CAS  PubMed  Google Scholar 

  19. Cheson BD, Bennett JM, Grever M, Kay N, Keating MJ, O'Brien S et al. National Cancer Institute-sponsored Working Group guidelines for chronic lymphocytic leukemia: revised guidelines for diagnosis and treatment. Blood 1996; 87: 4990–4997.

    CAS  PubMed  Google Scholar 

  20. Montserrat E, Sanchez-Bisono J, Vinolas N, Rozman C . Lymphocyte doubling time in chronic lymphocytic leukaemia: analysis of its prognostic significance. Br J Haematol 1986; 62: 567–575.

    Article  CAS  PubMed  Google Scholar 

  21. Ghia P, Guida G, Stella S, Gottardi D, Geuna M, Strola G et al. The pattern of CD38 expression defines a distinct subset of chronic lymphocytic leukemia (CLL) patients at risk of disease progression. Blood 2003; 15: 1262–1269.

    Article  Google Scholar 

  22. Krober A, Seiler T, Benner A, Bullinger L, Bruckle E, Lichter P et al. V(H) mutation status, CD38 expression level, genomic aberrations, and survival in chronic lymphocytic leukemia. Blood 2002; 15: 1410–1416.

    Google Scholar 

  23. Damle RN, Wasil T, Fais F, Ghiotto F, Valetto A, Allen SL et al. Ig V gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood 1999; 94: 1840–1847.

    CAS  PubMed  Google Scholar 

  24. Schroers R, Griesinger F, Trumper L, Haase D, Kulle B, Klein-Hitpass L et al. Combined analysis of ZAP-70 and CD38 expression as a predictor of disease progression in B-cell chronic lymphocytic leukemia. Leukemia 2005; 19: 750–758.

    Article  CAS  PubMed  Google Scholar 

  25. Gibbs G, Bromidge T, Howe D, Hopkins J, Johnson S . Comparison of flow cytometric methods for the measurement of ZAP-70 expression in a routine diagnostic laboratory. Clin Lab Haematol 2005; 27: 258–266.

    Article  CAS  PubMed  Google Scholar 

  26. Krober A, Bloehdorn J, Hafner S, Buhler A, Seiler T, Kienle D et al. Additional genetic high-risk features such as 11q deletion, 17p deletion, and V3-21 usage characterize discordance of ZAP-70 and VH mutation status in chronic lymphocytic leukemia. J Clin Oncol 2006; 24: 969–975.

    Article  PubMed  Google Scholar 

  27. Hanafelt TD, Jelinek D, Tschumper R, Schwager S, Nowakowski G, DeWald GW et al. Cytogenetic abnormalities can change during the course of the disease process in chronic lymphocytic leukemia. J Clin Oncol 2006; 24: 3218–3219.

    Article  Google Scholar 

  28. Sah SP, Matutes E, Wotherspoon AC, Morilla R, Catovsky D . A comparison of flow cytometry, bone marrow biopsy, and bone marrow aspirates in the detection of lymphoid infiltration in B cell disorders. J Clin Pathol 2003; 56: 129–132.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by Associazione Italiana per la Ricerca sul Cancro (AIRC), Milano, PRIN – MIUR, the CLL Global Research Foundation and Fondazione G. Berlucchi per la ricerca sul cancro.

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Authors and Affiliations

  1. Department of Clinical and Experimental Medicine, University of Verona, Verona, Italy

    R Zanotti, A Ambrosetti, F Zanetti, O Perbellini & G Pizzolo

  2. Department of Pathology, University of Verona, Verona, Italy

    M Lestani, A Remo, F Menestrina & M Chilosi

  3. Department of Oncology, Università Vita-Salute San Raffaele and Istituto Scientifico San Raffaele, Milano, Italy

    P Ghia & F Caligaris-Cappio

  4. Department of Medicine and Public Health, University of Verona, Verona, Italy

    C Pattaro

  5. Institute for Cancer Research and Treatment, IRCC, Candiolo e Ospedale Mauriziano ‘Umberto I’, Torino, Italy

    S Stella, G Prato, G Guida & F Caligaris-Cappio

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Correspondence to A Ambrosetti.

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Zanotti, R., Ambrosetti, A., Lestani, M. et al. ZAP-70 expression, as detected by immunohistochemistry on bone marrow biopsies from early-phase CLL patients, is a strong adverse prognostic factor. Leukemia 21, 102–109 (2007). https://doi.org/10.1038/sj.leu.2404458

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