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Acute Leukemias

Chimeric antigen receptors against CD33/CD123 antigens efficiently target primary acute myeloid leukemia cells in vivo

Abstract

As significant numbers of acute myeloid leukemia (AML) patients are still refractory to conventional therapies or experience relapse, immunotherapy using T cells expressing chimeric antigen receptors (CARs) might represent a valid treatment option. AML cells frequently overexpress the myeloid antigens CD33 and CD123, for which specific CARs can be generated. However, CD33 is also expressed on normal hematopoietic stem/progenitor cells (HSPCs), and its targeting could potentially impair normal hematopoiesis. In contrast, CD123 is widely expressed by AML, while low expression is detected on HSPCs, making it a much more attractive target. In this study we describe the in vivo efficacy and safety of using cytokine-induced killer (CIK) cells genetically modified to express anti-CD33 or anti-CD123 CAR to target AML. We show that both these modified T cells are very efficient in reducing leukemia burden in vivo, but only the anti-CD123 CAR has limited killing on normal HSPCs, thus making it a very attractive immunotherapeutic tool for AML treatment.

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References

  1. Majeti R . Monoclonal antibody therapy directly against human acute myeloid leukemia stem cells. Oncogene 2011; 30: 1009–1019.

    Article  CAS  Google Scholar 

  2. Horton S, Huntly B . Recent advances in acute myeloid leukemia stem cell biology. Haematologica 2012; 97: 966–974.

    Article  CAS  Google Scholar 

  3. Lu PH, Negrin RS . A novel population of expanded human CD3+CD56+ cells derived from T cells with potent in vivo antitumor activity in mice with severe combined immunodeficiency. J Immunol 1994; 153: 1687–1696.

    CAS  PubMed  Google Scholar 

  4. Franceschetti M, Pievani A, Borleri G, Vago L, Fleischhauer K, Golay J et al. Cytokine-induced killer cells are terminally differentiated activated CD8 cytotoxic T-EMRA lymphocytes. Exp Hematol 2009; 37: 616–628, e612.

    Article  CAS  Google Scholar 

  5. Linn YC, Lau SK, Liu BH, Ng LH, Yong HX, Hui KM . Characterization of the recognition and functional heterogeneity exhibited by cytokine-induced killer cell subsets against acute myeloid leukaemia target cell. Immunology 2009; 126: 423–435.

    Article  CAS  Google Scholar 

  6. Niam M, Linn YC, Fook Chong S, Lim TJ, Chu S, Choong A et al. Clinical scale expansion of cytokine-induced killer cells is feasible from healthy donors and patients with acute and chronic myeloid leukemia at various stages of therapy. Exp Hematol 2011; 39: 897–903, e891.

    Article  CAS  Google Scholar 

  7. Baker J, Verneris MR, Ito M, Shizuru JA, Negrin RS . Expansion of cytolytic CD8(+) natural killer T cells with limited capacity for graft-versus-host disease induction due to interferon gamma production. Blood 2001; 97: 2923–2931.

    Article  CAS  Google Scholar 

  8. Nishimura R, Baker J, Beilhack A, Zeiser R, Olson JA, Sega EI et al. In vivo trafficking and survival of cytokine-induced killer cells resulting in minimal GVHD with retention of antitumor activity. Blood 2008; 112: 2563–2574.

    Article  CAS  Google Scholar 

  9. Laport GG, Sheehan K, Baker J, Armstrong R, Wong RM, Lowsky R et al. NAdoptive immunotherapy with cytokine-induced killer cells for patients with relapse hematologic malignacies after allogeneic hematopoetic cell transplantation. Biol Blood Marrow Transplant 2011; 17: 1679–1687.

    Article  CAS  Google Scholar 

  10. Biagi E, Marin V, Attianese GM, Pizzitola I, Tettamanti S, Cribioli E et al. New advances in leukaemia immunotherapy by the use of Chimeric Artificial Antigen Receptors (CARs): state of the art and perspectives for the near future. Ital J Pediatr 2011; 37: 46.

    Article  CAS  Google Scholar 

  11. Kochenderfer JN, Rosenberg SA . Chimeric antigen receptor-modified T cells in CLL. N Engl J Med 2011; 365: 1937–1938, (author reply 1938).

    Article  CAS  Google Scholar 

  12. Grupp SA, Kalos M, Barrett D, Aplenc R, Porter DL, Rheingold SR et al. Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N Engl J Med 2013; 368: 1509–1518.

    Article  CAS  Google Scholar 

  13. Casucci M, Nicolis di Robilant B, Falcone L, Camisa B, Norelli M, Genovese P et al. CD44v6-targeted T cells mediate potent antitumor effects against acute myeloid leukemia and multiple myeloma. Blood 2013; 122: 3461–3472.

    Article  CAS  Google Scholar 

  14. Marin V, Pizzitola I, Agostoni V, Attianese GM, Finney H, Lawson A et al. Cytokine-induced killer cells for cell therapy of acute myeloid leukemia: improvement of their immune activity by expression of CD33-specific chimeric receptors. Haematologica 2010; 95: 2144–2152.

    Article  CAS  Google Scholar 

  15. Tettamanti S, Marin V, Pizzitola I, Magnani CF, Giordano Attianese GM, Cribioli E et al. Targeting of acute myeloid leukaemia by cytokine-induced killer cells redirected with a novel CD123-specific chimeric antigen receptor. Br J Haematol 2013; 161: 389–401.

    Article  CAS  Google Scholar 

  16. Mardiros A, Dos Santos C, McDonald T, Brown CE, Wang X, Budde LE et al. T cells expressing CD123-specific chimeric antigen receptors exhibit specific cytolytic effector functions and anti-tumor effects against human acute myeloid leukemia. Blood 2013; 122: 3138–3148.

    Article  CAS  Google Scholar 

  17. Jordan CT, Upchurch D, Szilvassy SJ, Guzman ML, Howard DS, Pettigrew AL et al. The interleukin-3 receptor alpha chain is a unique marker for human acute myelogenous leukemia stem cells. Leukemia 2000; 14: 1777–1784.

    Article  CAS  Google Scholar 

  18. Taussig DC, Pearce DJ, Simpson C, Rohatiner AZ, Lister TA, Kelly G et al. Hematopoietic stem cells express multiple myeloid markers: implications for the origin and targeted therapy of acute myeloid leukemia. Blood 2005; 106: 4086–4092.

    Article  CAS  Google Scholar 

  19. Jin L, Lee EM, Ramshaw HS, Busfield SJ, Peoppl AG, Wilkinson L et al. Monoclonal antibody-mediated targeting of CD123, IL-3 receptor alpha chain, eliminates human acute myeloid leukemic stem cells. Cell Stem Cell 2009; 5: 31–42.

    Article  CAS  Google Scholar 

  20. Sun Q, Woodcock JM, Rapoport A, Stromski FC, Korpelainen EJ, Bagley CJ et al. Monoclonal antibody 7G3 recognizes the N-terminal domain of the human interleukin-3 (IL-3) receptor alpha-chain and functions as a specific IL-3 receptor antagonist. Blood 1996; 87: 83–92.

    CAS  PubMed  Google Scholar 

  21. Vera J, Savoldo B, Vigouroux S, Biagi E, Pule M, Rossig C et al. T lymphocytes redirected against the kappa light chain of human immunoglobulin efficiently kill mature B lymphocyte-derived malignant cells. Blood 2006; 108: 3890–3897.

    Article  CAS  Google Scholar 

  22. Aigner M, Feulner J, Schaffer S, Kischel R, Kufer P, Schneider K et al. T lymphocytes can be effectively recruited for ex vivo and in vivo lysis of AML blasts by a novel CD33/CD3-bispecific BiTE((R)) antibody construct. Leukemia 2013; 7: 1107–1115.

    Article  Google Scholar 

  23. Walter RB, Appelbaum FR, Estey EH, Bernstein ID . Acute myeloid leukemia stem cells and CD33-targeted immunotherapy. Blood 2012; 119: 6198–6208.

    Article  CAS  Google Scholar 

  24. Testa U, Riccioni R, Militi S, Coccia E, Stellacci E, Samoggia P et al. Elevated expression of IL3Ralpha in acute myeloid leukemia associated with enhanced blast proliferation, increased cellularity, and poor prognosis. Blood 2002; 100: 2980–2988.

    Article  CAS  Google Scholar 

  25. Hernández-Caselles T, Martínez-Esparza M, Pérez-Oliva AB, Quintanilla-Cecconi AM, García-Alonso A, Alvarez-López DM et al. A study of CD33 (SIGLEC-3) antigen expression and function on activated human T and NK cells: two isoforms of CD33 are generated by alternative splicing. J Leukoc Biol 2006; 79: 46–58.

    Article  Google Scholar 

  26. Sievers EL, Larson RA, Stadtmauer EA, Estey E, Löwenberg B, Dombret H et alMylotarg Study Group. Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukemia in first relapse. J Clin Oncol 2001; 19: 3244–3254.

    Article  CAS  Google Scholar 

  27. Roberts AW, He S, Bradstock KF, Hertzberg MS, Durrant STS, Ritchie D et al. A phase 1 and correlative biological study of CSL360 (anti-CD123 mAb) in AML. Blood 2008; 112, (2956a).

  28. Kugler M, Stein C, Kellner C, Mentz K, Saul D, Schwenkert M et al. A recombinant trispecific single chain Fv derivative directed against CD123 and CD33 mediates effective elimination of acute myeloid leukemia cells by dual targeting. Br J Haematol 2010; 150: 574–586.

    Article  Google Scholar 

  29. Stein C, Kellner C, Kugler M, Reiff N, Mentz K, Schwenkert M et al. Novel conjugates of single -chain Fv antibody fragments specific for stem cell antigen CD1233 mediates a potent death of acute myeloid leukemia. Br J Haematol 2010; 148: 879–889.

    Article  CAS  Google Scholar 

  30. Cartellieri M, Bachmann M, Feldmann A, Bippes C, Stamova S, Wehner R et al. Chimeric antigen receptor-engineered T cells for immunotherapy of cancer. J Biomed Biotechnol 2010; 2010: 956304.

    Article  Google Scholar 

  31. Huang X, Guo H, Kang J, Choi S, Zhou TC, Tammana S et al. Sleeping beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies. Mol Ther 2008; 16: 580–589.

    Article  CAS  Google Scholar 

  32. Barrett DM, Liu X, Jiang S, June CH, Grupp SA, Zhao Y . Regimen-specific effects of RNA-modified chimeric antigen receptor T cells in mice with advance leukemia. Hum Gene Ther 2013; 24: 717–727.

    Article  CAS  Google Scholar 

  33. Kebriaei HHP, Singh H, Olivares S, Figliola M, Dawson M, Jena B, Jackson DBR, Rondon G, McNiece I, Hackett P, Shpall E, Champlin RE . First clinical trials employing Sleeping Beauty system and artificial chimeric antigen presenting cells to generate T cells expressing CD-19 specific chimeric antigen receptor. 18th Congress of the European Hematology Association.. Stockholm, Sweden, 2013.

    Google Scholar 

  34. Di Stasi A, Tey SK, Dotti G, Fujita Y, Kennedy-Nasser A, Martinez C et al. Inducible apoptosis as a safety switch for adoptive cell therapy. N Engl J Med 2011; 365: 1673–1683.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Dr Alfredo Pagliuca for providing us with the Luciferase constructs. We would like to thanks Drs Gino Vairo and Angel Lopez from CSL Limited Australia, for providing us with the initial CD123 mAb from which we derived the CAR. This work was funded in part by Cancer Research UK to DB and by Fondazione Tettamanti and Associazione Italiana per la Ricerca sul Cancro (AIRC) (AIRC 5 × 1000; AIRC-IG8666) to AB.

Author contributions

IP designed and coordinated the research, performed experiments, analyzed the data and wrote the paper. FAA performed the experiments, interpreted the results, revised the data and wrote the paper. KRP and FL performed the experiments. ST and OS provided AML samples and performed the experiments. AB, EB and BD designed and coordinated the research, revised the data and wrote the paper.

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Correspondence to A Biondi.

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Pizzitola, I., Anjos-Afonso, F., Rouault-Pierre, K. et al. Chimeric antigen receptors against CD33/CD123 antigens efficiently target primary acute myeloid leukemia cells in vivo. Leukemia 28, 1596–1605 (2014). https://doi.org/10.1038/leu.2014.62

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