Abstract
Patients with chemo-refractory acute myeloid leukemia (AML) have a dismal prognosis. Chimeric antigen receptor T (CART) cell therapy has produced exciting results in CD19+ malignancies and may overcome many of the limitations of conventional leukemia therapies. We developed CART cells to target CD33 (CART33) using the anti-CD33 single chain variable fragment used in gemtuzumab ozogamicin (clone My96) and tested the activity and toxicity of these cells. CART33 exhibited significant effector functions in vitro and resulted in eradication of leukemia and prolonged survival in AML xenografts. CART33 also resulted in human lineage cytopenias and reduction of myeloid progenitors in xenograft models of hematopoietic toxicity, suggesting that permanently expressed CD33-specific CART cells would have unacceptable toxicity. To enhance the viability of CART33 as an option for AML, we designed a transiently expressed mRNA anti-CD33 CAR. Gene transfer was carried out by electroporation into T cells and resulted in high-level expression with potent but self-limited activity against AML. Thus our preclinical studies show potent activity of CART33 and indicate that transient expression of anti-CD33 CAR by RNA modification could be used in patients to avoid long-term myelosuppression. CART33 therapy could be used alone or as part of a preparative regimen prior to allogeneic transplantation in refractory AML.
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References
Siegel R, Ma J, Zou Z, Jemal A . Cancer statistics, 2014. CA Cancer J Clin 2014; 64: 9–29.
Thein MS, Ershler WB, Jemal A, Yates JW, Baer MR . Outcome of older patients with acute myeloid leukemia: an analysis of SEER data over 3 decades. Cancer 2013; 119: 2720–2727.
Hamadani M, Awan FT, Copelan EA . Hematopoietic stem cell transplantation in adults with acute myeloid leukemia. Biol Blood Marrow Transplant 2008; 14: 556–567.
Duval M, Klein JP, He W, Cahn JY, Cairo M, Camitta BM et al. Hematopoietic stem-cell transplantation for acute leukemia in relapse or primary induction failure. J Clin Oncol 2010; 28: 3730–3738.
Estey EH . Acute myeloid leukemia: 2013 update on risk-stratification and management. Am J Hematol 2013; 88: 318–327.
Kalos M, June CH . Adoptive T cell transfer for cancer immunotherapy in the era of synthetic biology. Immunity 2013; 39: 49–60.
Gross G, Waks T, Eshhar Z . Expression of immunoglobulin-T-cell receptor chimeric molecules as functional receptors with antibody-type specificity. Proc Natl Acad Sci USA 1989; 86: 10024–10028.
Porter DL, Levine BL, Kalos M, Bagg A, June CH . Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N Engl J Med 2011; 365: 725–733.
Grupp SA, Kalos M, Barrett D, Aplenc R, Porter D, Rheingold S et al. Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N Engl J Med 2013; 368: 1509–1518.
Kalos M, Levine BL, Porter DL, Katz S, Grupp SA, Bagg A et al. T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced leukemia. Sci Transl Med 2011; 3: 95ra73.
Kochenderfer JN, Dudley ME, Feldman SA, Wilson WH, Spaner DE, Maric I et al. B-cell depletion and remissions of malignancy along with cytokine-associated toxicity in a clinical trial of anti-CD19 chimeric-antigen-receptor-transduced T cells. Blood 2012; 119: 2709–2720.
Davila ML, Riviere I, Wang X, Bartido S, Park J, Curran K et al. Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia. Sci Transl Med 2014; 6: 224ra225.
Kochenderfer JN, Dudley ME, Kassim SH, Somerville RP, Carpenter RO, Stetler-Stevenson M et al. Chemotherapy-refractory diffuse large B-cell lymphoma and indolent B-cell malignancies can be effectively treated with autologous T cells expressing an anti-CD19 chimeric antigen receptor. J Clin Oncol 2014; 33: 540–549.
Maude SL, Frey N, Shaw PA, Aplenc R, Barrett DM, Bunin NJ et al. Chimeric antigen receptor T cells for sustained remissions in leukemia. N Engl J Med 2014; 371: 1507–1517.
Cruz CR, Micklethwaite KP, Savoldo B, Ramos CA, Lam S, Ku S et al. Infusion of donor-derived CD19-redirected virus-specific T cells for B-cell malignancies relapsed after allogeneic stem cell transplant: a phase 1 study. Blood 2013; 122: 2965–2973.
Walter RB, Gooley TA, van der Velden VH, Loken MR, van Dongen JJ, Flowers DA et al. CD33 expression and P-glycoprotein-mediated drug efflux inversely correlate and predict clinical outcome in patients with acute myeloid leukemia treated with gemtuzumab ozogamicin monotherapy. Blood 2007; 109: 4168–4170.
Griffin JD, Linch D, Sabbath K, Larcom P, Schlossman SF . A monoclonal antibody reactive with normal and leukemic human myeloid progenitor cells. Leuk Res 1984; 8: 521–534.
Dinndorf PA, Andrews RG, Benjamin D, Ridgway D, Wolff L, Bernstein ID . Expression of normal myeloid-associated antigens by acute leukemia cells. Blood 1986; 67: 1048–1053.
Schwonzen M, Diehl V, Dellanna M, Staib P . Immunophenotyping of surface antigens in acute myeloid leukemia by flow cytometry after red blood cell lysis. Leuk Res 2007; 31: 113–116.
Hoyer JD, Grogg KL, Hanson CA, Gamez JD, Dogan A . CD33 detection by immunohistochemistry in paraffin-embedded tissues: a new antibody shows excellent specificity and sensitivity for cells of myelomonocytic lineage. Am J Clin Pathol 2008; 129: 316–323.
Hills RK, Castaigne S, Appelbaum FR, Delaunay J, Petersdorf S, Othus M et al. Addition of gemtuzumab ozogamicin to induction chemotherapy in adult patients with acute myeloid leukaemia: a meta-analysis of individual patient data from randomised controlled trials. Lancet Oncol 2014; 15: 986–996.
Gill S, Tasian SK, Ruella M, Shestova O, Li Y, Porter DL et al. Preclinical targeting of human acute myeloid leukemia and myeloablation using chimeric antigen receptor-modified T cells. Blood 2014; 123: 2343–2354.
Milone MC, Fish JD, Carpenito C, Carroll RG, Binder GK, Teachey D et al. Chimeric receptors containing CD137 signal transduction domains mediate enhanced survival of T cells and increased antileukemic efficacy in vivo. Mol Ther 2009; 17: 1453–1464.
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 advanced leukemia. Hum Gene Ther 2013; 24: 717–727.
Barrett DM, Zhao Y, Liu X, Jiang S, Carpenito C, Kalos M et al. Treatment of advanced leukemia in mice with mRNA engineered T cells. Hum Gene Ther 2011; 22: 1575–1586.
Zhao Y, Zheng Z, Cohen CJ, Gattinoni L, Palmer DC, Restifo NP et al. High-efficiency transfection of primary human and mouse T lymphocytes using RNA electroporation. Mol Ther 2006; 13: 151–159.
Betts MR, Koup RA . Detection of T-cell degranulation: CD107a and b. Methods Cell Biol 2004; 75: 497–512.
Cao LF, Krymskaya L, Tran V, Mi S, Jensen MC, Blanchard S et al. Development and application of a multiplexable flow cytometry-based assay to quantify cell-mediated cytolysis. Cytometry A 2010; 77: 534–545.
Pollard JA, Alonzo TA, Loken M, Gerbing RB, Ho PA, Bernstein ID et al. Correlation of CD33 expression level with disease characteristics and response to gemtuzumab ozogamicin containing chemotherapy in childhood AML. Blood 2012; 119: 3705–3711.
Walter RB, Raden BW, Kamikura DM, Cooper JA, Bernstein ID . Influence of CD33 expression levels and ITIM-dependent internalization on gemtuzumab ozogamicin-induced cytotoxicity. Blood 2005; 105: 1295–1302.
Ehninger A, Kramer M, Rollig C, Thiede C, Bornhauser M, von Bonin M et al. Distribution and levels of cell surface expression of CD33 and CD123 in acute myeloid leukemia. Blood Cancer J 2014; 4: e218.
Carpenito C, Milone MC, Hassan R, Simonet JC, Lakhal M, Suhoski MM et al. Control of large, established tumor xenografts with genetically retargeted human T cells containing CD28 and CD137 domains. Proc Natl Acad Sci USA 2009; 106: 3360–3365.
Wunderlich M, Chou FS, Link KA, Mizukawa B, Perry RL, Carroll M et al. AML xenograft efficiency is significantly improved in NOD/SCID-IL2RG mice constitutively expressing human SCF, GM-CSF and IL-3. Leukemia 2010; 24: 1785–1788.
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.
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 antitumor effects against human acute myeloid leukemia. Blood 2013; 122: 3138–3148.
Pizzitola I, Anjos-Afonso F, Rouault-Pierre K, Lassailly F, Tettamanti S, Spinelli O et al. Chimeric antigen receptors against CD33/CD123 antigens efficiently target primary acute myeloid leukemia cells in vivo. Leukemia 2014; 28: 1596–1605.
Dutour A, Marin V, Pizzitola I, Valsesia-Wittmann S, Lee D, Yvon E et al. In vitro and in vivo antitumor effect of anti-CD33 chimeric receptor-expressing EBV-CTL against CD33 acute myeloid leukemia. Adv Hematol 2012; 2012: 683065.
Hudecek M, Sommermeyer D, Kosasih PL, Silva-Benedict A, Liu L, Rader C et al. The non-signaling extracellular spacer domain of chimeric antigen receptors is decisive for in vivo antitumor activity. Cancer Immunol Res 2014; 3: 125–135.
Haso W, Lee DW, Shah NN, Stetler-Stevenson M, Yuan CM, Pastan IH et al. Anti-CD22-chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood 2013; 121: 1165–1174.
Larson RA, Sievers EL, Stadtmauer EA, Lowenberg B, Estey EH, Dombret H et al. Final report of the efficacy and safety of gemtuzumab ozogamicin (Mylotarg) in patients with CD33-positive acute myeloid leukemia in first recurrence. Cancer 2005; 104: 1442–1452.
Kantarjian H, Thomas D, Jorgensen J, Kebriaei P, Jabbour E, Rytting M et al. Results of inotuzumab ozogamicin, a CD22 monoclonal antibody, in refractory and relapsed acute lymphocytic leukemia. Cancer 2013; 119: 2728–2736.
Kantarjian H, Thomas D, Jorgensen J, Jabbour E, Kebriaei P, Rytting M et al. Inotuzumab ozogamicin, an anti-CD22-calecheamicin conjugate, for refractory and relapsed acute lymphocytic leukaemia: a phase 2 study. Lancet Oncol 2012; 13: 403–411.
Ritchie DS, Neeson PJ, Khot A, Peinert S, Tai T, Tainton K et al. Persistence and efficacy of second generation CAR-T cell against the LeY antigen in acute myeloid leukemia. Mol Ther 2013; 21: 2122–2129.
Wang QS, Wang Y, Lv HY, Han QW, Fan H, Guo B et al. Treatment of CD33-directed chimeric antigen receptor-modified T cells in one patient with relapsed and refractory acute myeloid leukemia. Mol Ther 2014; 23: 184–191.
Beatty GL, Haas AR, Maus MV, Torigian DA, Soulen MC, Plesa G et al. Mesothelin-specific chimeric antigen receptor mRNA-engineered T cells induce anti-tumor activity in solid malignancies. Cancer Immunol Res 2014; 2: 112–120.
Bonini C, Bondanza A, Perna SK, Kaneko S, Traversari C, Ciceri F et al. The suicide gene therapy challenge: how to improve a successful gene therapy approach. Mol Ther 2007; 15: 1248–1252.
Acknowledgements
This work was funded by a research agreement with Novartis Pharmaceuticals, a Leukemia and Lymphoma Society Specialized Centers of Research grant to CHJ, a National Institutes of Health T32 award to MK (T32-GM008076) and an American Society of Hematology Scholar award to SG. We thank Fang Chen and Simon Lacey for performing Luminex assay. Imaging was performed at the University of Pennsylvania Small Animal Imaging Facility (SAIF) Optical/Bioluminescence Core, supported by NIH grant CA016520.
Author Contributions
SSK designed and performed research, analyzed data and wrote the paper; MR and SG designed, performed research and wrote the paper; OS performed research, MK, VA, JM and DS performed research; JS contributed reagents and performed research; DLP designed research and wrote the paper; MC designed research and contributed reagents; and CHJ designed research, contributed reagents and wrote the paper. All authors read and approved the manuscript.
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CHJ and DLP have filed patent applications related to CAR technology and could potentially receive licensing royalties from Novartis corporation. The other authors declare no conflict of interest.
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Kenderian, S., Ruella, M., Shestova, O. et al. CD33-specific chimeric antigen receptor T cells exhibit potent preclinical activity against human acute myeloid leukemia. Leukemia 29, 1637–1647 (2015). https://doi.org/10.1038/leu.2015.52
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DOI: https://doi.org/10.1038/leu.2015.52
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