Skip to main content

Advertisement

Log in

Natural killer cells in acute myeloid leukemia patients: from phenotype to transcriptomic analysis

  • Original Article
  • Published:
Immunologic Research Aims and scope Submit manuscript

Abstract

Chemotherapies allow complete remission in more than 50 % of patients with acute myeloid leukemia (AML), however, with frequent relapse. This suggests that residual leukemic cells may escape to chemotherapy and immune system. Natural killer (NK) cells from AML patients (AML-NK) have a weaker natural cytotoxicity-activating receptors (NCRs) expression than NK cells from healthy donors (HD-NK). Coding genes for NCR1/NKp46, NCR2/NKp44 and NCR3/NKp30 are located at different loci on two different chromosomes; however, their expression is tightly coordinated. Most NK cells express either high (NCRbright) or low levels (NCRdull) of all three NCRs. This suggests the existence of negative/positive regulation factor(s) common to the three receptors. In order to find transcription factor(s) or pathway(s) involved in NCRs co-regulation, this study compared the transcriptomic signature of HD-NK and AML-NK cells, before and after in vitro NK cells culture. Microarrays analysis revealed a specific NK cells transcriptomic signature in patients with AML. However, in vitro NK cells expansion erased this signature and up-regulated expression of central molecules of NK functions, such as NCR, NKG2D and also ETS-1, regardless of their origin, i.e., AML-NK vs HD-NK. ETS-1 transcription factor was shown to bind to a specific and common region in the NCRs promoters, thus appearing as a good candidate to explain the coordinated regulation of three NCRs. Such results are encouraging regarding in vitro AML-NK cytotoxicity restoration and provide a new conceptual support for innovative cellular therapy based on in vitro NK cells expansion before their reinfusion in AML patients.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

CR:

Complete remission

NK:

Natural killer

AML:

Acute myeloid leukemia

TF:

Transcription factor

NCR:

Natural cytotoxicity receptor

EBV:

Epstein barr virus

PBMC:

Peripheral blood mononuclear cells

p:

p values

FC:

Fold change

TFBS:

Transcription factor binding sites

References

  1. Döhner H, Estey EH, Amadori S, Appelbaum FR, Büchner T, Burnett AK, et al. Diagnosis and management of acute myeloid leukemia in adults: recommendations from an international expert panel, on behalf of the European LeukemiaNet. Blood. 2010;115:453–74.

    Article  PubMed  Google Scholar 

  2. Jemal A, Clegg LX, Ward E, Ries LAG, Wu X, Jamison PM, et al. Annual report to the nation on the status of cancer, 1975–2001, with a special feature regarding survival. Cancer. 2004;101:3–27.

    Article  PubMed  Google Scholar 

  3. Cheson BD, Bennett JM, Kopecky KJ, Büchner T, Willman CL, Estey EH, et al. Revised recommendations of the international working group for diagnosis, standardization of response criteria, treatment outcomes, and reporting standards for therapeutic trials in acute myeloid leukemia. J Clin Oncol Off J Am Soc Clin Oncol. 2003;21:4642–9.

    Article  Google Scholar 

  4. Asai O, Longo DL, Tian ZG, Hornung RL, Taub DD, Ruscetti FW, et al. Suppression of graft-versus-host disease and amplification of graft-versus-tumor effects by activated natural killer cells after allogeneic bone marrow transplantation. J Clin Invest. 1998;101:1835–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Jiang YZ, Barrett AJ, Goldman JM, Mavroudis DA. Association of natural killer cell immune recovery with a graft-versus-leukemia effect independent of graft-versus-host disease following allogeneic bone marrow transplantation. Ann Hematol. 1997;74:1–6.

    Article  CAS  PubMed  Google Scholar 

  6. Lavigne P, Acres S. Advisory committee on epidemiology, resolution. Can Dis Wkly Rep Rapp Hebd Mal Au Can. 1989;15:127–9.

    CAS  Google Scholar 

  7. Elkins WL, Pickard A, Pierson GR. Deficient expression of class-I HLA in some cases of acute leukemia. Cancer Immunol Immunother CII. 1984;18:91–100.

    Article  CAS  PubMed  Google Scholar 

  8. Urlacher A, Falkenrodt A, Tongio MM, Mayer S. HLA class I antigens on normal and leukemic cells (quantitative analysis). Tissue Antigens. 1987;29:237–45.

    Article  CAS  PubMed  Google Scholar 

  9. Costello RT, Sivori S, Marcenaro E, Lafage-Pochitaloff M, Mozziconacci M-J, Reviron D, et al. Defective expression and function of natural killer cell-triggering receptors in patients with acute myeloid leukemia. Blood. 2002;99:3661–7.

    Article  CAS  PubMed  Google Scholar 

  10. Costello RT, Knoblauch B, Sanchez C, Mercier D, Le Treut T, Sébahoun G. Expression of natural killer cell activating receptors in patients with chronic lymphocytic leukaemia. Immunology. 2012;135:151–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Fauriat C, Just-Landi S, Mallet F, Arnoulet C, Sainty D, Olive D, et al. Deficient expression of NCR in NK cells from acute myeloid leukemia: evolution during leukemia treatment and impact of leukemia cells in NCRdull phenotype induction. Blood. 2007;109:323–30.

    Article  CAS  PubMed  Google Scholar 

  12. Kiladjian J-J, Bourgeois E, Lobe I, Braun T, Visentin G, Bourhis J-H, et al. Cytolytic function and survival of natural killer cells are severely altered in myelodysplastic syndromes. Leukemia. 2006;20:463–70.

    Article  CAS  PubMed  Google Scholar 

  13. Costello RT, Boehrer A, Sanchez C, Mercier D, Baier C, Le Treut T, et al. Differential expression of natural killer cell activating receptors in blood versus bone marrow in patients with monoclonal gammopathy. Immunology. 2013;139:338–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Costello RT, Knoblauch B, Sanchez C, Mercier D, Le Treut T, Sébahoun G. Expression of natural killer cell activating receptors in patients with chronic lymphocytic leukaemia. Immunology. 2012;135:151–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Martner A, Rydström A, Riise RE, Aurelius J, Brune M, Foà R, et al. NK cell expression of natural cytotoxicity receptors may determine relapse risk in older AML patients undergoing immunotherapy for remission maintenance. Oncotarget. 2015;6:42569–74.

    PubMed  PubMed Central  Google Scholar 

  16. Martner A, Rydström A, Riise RE, Aurelius J, Anderson H, Brune M, et al. Role of natural killer cell subsets and natural cytotoxicity receptors for the outcome of immunotherapy in acute myeloid leukemia. Oncoimmunology. 2016;5:e1041701.

    Article  PubMed  Google Scholar 

  17. Brune M, Hansson M, Mellqvist UH, Hermodsson S, Hellstrand K. NK cell-mediated killing of AML blasts: role of histamine, monocytes and reactive oxygen metabolites. Eur J Haematol. 1996;57:312–9.

    Article  CAS  PubMed  Google Scholar 

  18. Thorén FB, Romero AI, Brune M, Hellstrand K. Histamine dihydrochloride and low-dose interleukin-2 as post-consolidation immunotherapy in acute myeloid leukemia. Expert Opin Biol Ther. 2009;9:1217–23.

    Article  PubMed  Google Scholar 

  19. Ramirez K, Chandler KJ, Spaulding C, Zandi S, Sigvardsson M, Graves BJ, et al. Gene deregulation and chronic activation in natural killer cells deficient in the transcription factor ETS1. Immunity. 2012;36:921–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Reimand J, Kull M, Peterson H, Hansen J, Vilo J. g:Profiler—a web-based toolset for functional profiling of gene lists from large-scale experiments. Nucleic Acid Res. 2007;35(suppl 2):W193–200.

    Article  PubMed  PubMed Central  Google Scholar 

  21. van Helden J, André B, Collado-Vides J. Extracting regulatory sites from the upstream region of yeast genes by computational analysis of oligonucleotide frequencies. J Mol Biol. 1998;281:827–42.

    Article  PubMed  Google Scholar 

  22. Turatsinze J-V, Thomas-Chollier M, Defrance M, van Helden J. Using RSAT to scan genome sequences for transcription factor binding sites and cis-regulatory modules. Nat Protoc. 2008;3:1578–88.

    Article  CAS  PubMed  Google Scholar 

  23. Thomas-Chollier M, Defrance M, Medina-Rivera A, Sand O, Herrmann C, Thieffry D, et al. RSAT 2011: regulatory sequence analysis tools. Nucleic Acids Res. 2011;39:W86–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Barton K, Muthusamy N, Fischer C, Ting CN, Walunas TL, Lanier LL, et al. The Ets-1 transcription factor is required for the development of natural killer cells in mice. Immunity. 1998;9:555–63.

    Article  CAS  PubMed  Google Scholar 

  25. Lacorazza HD, Miyazaki Y, Di Cristofano A, Deblasio A, Hedvat C, Zhang J, et al. The ETS protein MEF plays a critical role in perforin gene expression and the development of natural killer and NK-T cells. Immunity. 2002;17:437–49.

    Article  CAS  PubMed  Google Scholar 

  26. Townsend MJ, Weinmann AS, Matsuda JL, Salomon R, Farnham PJ, Biron CA, et al. T-bet regulates the terminal maturation and homeostasis of NK and Valpha14i NKT cells. Immunity. 2004;20:477–94.

    Article  CAS  PubMed  Google Scholar 

  27. Wang JH, Nichogiannopoulou A, Wu L, Sun L, Sharpe AH, Bigby M, et al. Selective defects in the development of the fetal and adult lymphoid system in mice with an Ikaros null mutation. Immunity. 1996;5:537–49.

    Article  CAS  PubMed  Google Scholar 

  28. Glimcher LH, Townsend MJ, Sullivan BM, Lord GM. Recent developments in the transcriptional regulation of cytolytic effector cells. Nat Rev Immunol. 2004;4:900–11.

    Article  CAS  PubMed  Google Scholar 

  29. Koizumi H, Horta MF, Youn BS, Fu KC, Kwon BS, Young JD, et al. Identification of a killer cell-specific regulatory element of the mouse perforin gene: an Ets-binding site-homologous motif that interacts with Ets-related proteins. Mol Cell Biol. 1993;13:6690–701.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Khaznadar Z, Boissel N, Agaugué S, Henry G, Cheok M, Vignon M, et al. Defective NK cells in acute myeloid leukemia patients at diagnosis are associated with blast transcriptional signatures of immune evasion. J Immunol. 1950;2015(195):2580–90.

    Google Scholar 

Download references

Acknowledgments

We would like to thank INSERM, AP-HM and Aix-Marseille University for grants. We would like to thanks Martin VILLALBA who gave us graciously EBV cell line. We are grateful to the patients who gave their informed consent to the use of their samples for research. We are grateful to the IBiSA Transcriptomics and Genomics Marseille-Luminy (TGML) platform were supported by the France Génomique National infrastructure. We thank Laurence Borge for assistance and the use of the cell culture platform facilities (CRCM U1068, Marseille) Plateforme de Culture Cellulaire de Marseille-Luminy, bâtiment TPR2, 163 Avenue de Luminy, 13009 Marseille. We thank also Geneviève Victorero for technical supports. This work was supported by grants from AP-HM, Assistance Publique—Hôpitaux de Marseille, (AORC APHM Junior), and Aix-Marseille Université, TGML platform supported by the France Génomique.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R. Costello.

Ethics declarations

Conflict of interest

All authors declare no financial conflict of interest.

Additional information

G. Venton and Y. Labiad have contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Venton, G., Labiad, Y., Colle, J. et al. Natural killer cells in acute myeloid leukemia patients: from phenotype to transcriptomic analysis. Immunol Res 64, 1225–1236 (2016). https://doi.org/10.1007/s12026-016-8848-0

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12026-016-8848-0

Keywords

Navigation