Tumor immunotherapy based on the use of chimeric antigen receptor modified T cells (CAR T cells) is a promising approach for the treatment of refractory hematological malignancies. However, a robust response mediated by CAR T cells is observed only in a minority of patients and the expansion and persistence of CAR T cells in vivo is mostly unpredictable.Lenalidomide (LEN) is an immunomodulatory drug currently approved for the treatment of multiple myeloma (MM) and mantle cell lymphoma, while it is clinically tested in the therapy of diffuse large B-cell lymphoma of activated B cell immunophenotype. LEN was shown to increase antitumor immune responses at least partially by modulating the activity of E3 ubiquitin ligase Cereblon, which leads to increased ubiquitinylation of Ikaros and Aiolos transcription factors, which in turn results in changed expression of various receptors on the surface of tumor cells. In order to enhance the effectiveness of CAR-based immunotherapy, we assessed the anti-lymphoma efficacy of LEN in combination with CAR19 T cells or CAR20 T cells in vitro and in vivo using various murine models of aggressive B-cell non-Hodgkin lymphomas (B-NHL).Immunodeficient NSG mice were transplanted with various human B-NHL cells followed by treatment with CAR19 or CAR20 T cells with or without LEN. Next, CAR19 T cells were subjected to series of tests in vitro to evaluate their response and signaling capacity following recognition of B cell in the presence or absence of LEN.Our data shows that LEN significantly enhances antitumor functions of CAR19 and CAR20 T cells in vivo. Additionally, it enhances production of interferon gamma by CAR19 T cells and augments cell signaling via CAR19 protein in T cells in vitro. Our data further suggests that LEN works through direct effects on T cells but not on B-NHL cells. The biochemical events underlying this costimulatory effect of LEN are currently being investigated. In summary, our data supports the use of LEN for augmentation of CAR-based immunotherapy in the clinical grounds.

Department of Hematology Charles University General Hospital Prague Czech Republic

Department of Hematology Charles University General Hospital Prague Czech Republic; Institute of Molecular Genetics Czech Academy of Sciences Prague Czech Republic

Department of Hematology Charles University General Hospital Prague Czech Republic; Institute of Pathological Physiology 1st Faculty of Medicine Charles University Prague Czech Republic

Institute of Molecular Genetics Czech Academy of Sciences Prague Czech Republic

Institute of Pathological Physiology 1st Faculty of Medicine Charles University Prague Czech Republic

Zobrazit více v PubMed

Maude SL, Frey N, Shaw PA, Aplenc R, Barrett DM, Bunin NJ, Chew A, Gonzalez VE, Zheng Z, Lacey SF et al., Chimeric antigen receptor T cells for sustained remissions in leukemia. N Engl J Med 2014; 371(16):1507-17; PMID:25317870; http://dx.doi.org/ 10.1056/NEJMoa1407222 PubMed DOI PMC

Maher J. Immunotherapy of malignant disease using chimeric antigen receptor engrafted T cells. ISRN Oncol 2012; 2012:278093; PMID:23304553; http://dx.doi.org/ 10.5402/2012/278093 PubMed DOI PMC

Curran KJ, Seinstra BA, Nikhamin Y, Yeh R, Usachenko Y, van Leeuwen DG, Purdon T, Pegram HJ, Brentjens RJ. Enhancing Antitumor Efficacy of Chimeric Antigen Receptor T Cells Through Constitutive CD40L Expression. Mol Ther 2015; 23(4):769-78; PMID:25582824; http://dx.doi.org/ 10.1038/mt.2015.4 PubMed DOI PMC

Han EQ, Li XL, Wang CR, Li TF, Han SY. Chimeric antigen receptor-engineered T cells for cancer immunotherapy: progress and challenges. J Hematol Oncol 2013; 6:47; PMID:23829929; PMID:24353912; http://dx.doi.org/20942636 10.1186/1756-8722-6-47 PubMed DOI PMC

John LB, Kershaw MH, Darcy PK. Blockade of PD-1 immunosuppression boosts CAR T-cell therapy. Oncoimmunology 2013; 2(10):e26286; PMID:24353912; http://dx.doi.org/20942636 10.4161/onci.26286 PubMed DOI PMC

Heise C, Carter T, Schafer P, Chopra R. Pleiotropic mechanisms of action of lenalidomide efficacy in del(5q) myelodysplastic syndromes. Expert Rev Anticancer Ther 2010; 10 (10):1663-72; PMID:20942636; http://dx.doi.org/ 10.1586/era.10.135 PubMed DOI

Kronke J, Udeshi ND, Narla A, Grauman P, Hurst SN, McConkey M, Svinkina T, Heckl D, Comer E, Li X et al., Lenalidomide causes selective degradation of IKZF1 and IKZF3 in multiple myeloma cells. Science 2014; 343(6168): 301-5; PMID:24292625; http://dx.doi.org/ 10.1126/science.1244851 PubMed DOI PMC

Gandhi AK, Kang J, Havens CG, Conklin T, Ning Y, Wu L, Ito T, Ando H, Waldman MF, Thakurta A et al., Immunomodulatory agents lenalidomide and pomalidomide co-stimulate T cells by inducing degradation of T cell repressors Ikaros and Aiolos via modulation of the E3 ubiquitin ligase complex CRL4(CRBN). Br J Haematol 2014; 164(6):811-21; PMID:24328678; http://dx.doi.org/ 10.1111/bjh.12708 PubMed DOI PMC

LeBlanc R, Hideshima T, Catley LP, Shringarpure R, Burger R, Mitsiades N, Mitsiades C, Cheema P, Chauhan D, Richardson PG et al., Immunomodulatory drug costimulates T cells via the B7-CD28 pathway. Blood 2004; 103(5):1787-90; PMID:14512311; http://dx.doi.org/ 10.1182/blood-2003-02-0361 PubMed DOI

Schafer PH, Gandhi AK, Loveland MA, Chen RS, Man HW, Schnetkamp PP, Wolbring G, Govinda S, Corral LG, Payvandi F et al., Enhancement of cytokine production and AP-1 transcriptional activity in T cells by thalidomide-related immunomodulatory drugs. J Pharmacol Exp Ther 2003; 305(3):1222-32; PMID:12649301; http://dx.doi.org/ 10.1124/jpet.102.048496 PubMed DOI

Haslett PA, Corral LG, Albert M, Kaplan G. Thalidomide costimulates primary human T lymphocytes, preferentially inducing proliferation, cytokine production, and cytotoxic responses in the CD8+ subset. J Exp Med 1998; 187(11):1885-92; PMID:9607928; http://dx.doi.org/ 10.1084/jem.187.11.1885 PubMed DOI PMC

Klanova M, Soukup T, Jaksa R, Molinsky J, Lateckova L, Maswabi BC, Prukova D, Brezinova J, Michalova K, Vockova P et al., Mouse models of mantle cell lymphoma, complex changes in gene expression and phenotype of engrafted MCL cells: implications for preclinical research. Lab Invest 2014; 94(7):806-17; PMID:24862967; http://dx.doi.org/ 10.1038/labinvest.2014.61 PubMed DOI

Verhelle D, Corral LG, Wong K, Mueller JH, Moutouh-de Parseval L, Jensen-Pergakes K, Schafer PH, Chen R, Glezer E, Ferguson GD et al., Lenalidomide and CC-4047 inhibit the proliferation of malignant B cells while expanding normal CD34+ progenitor cells. Cancer Res 2007; 67(2):746-55; PMID:17234786; http://dx.doi.org/ 10.1158/0008-5472.CAN-06-2317 PubMed DOI

Gorgun G, Calabrese E, Soydan E, Hideshima T, Perrone G, Bandi M, Cirstea D, Santo L, Hu Y, Tai YT et al., Immunomodulatory effects of lenalidomide and pomalidomide on interaction of tumor and bone marrow accessory cells in multiple myeloma. Blood 2010; 116 (17):3227-37; PMID:20651070; http://dx.doi.org/ 10.1182/blood-2010-04-279893 PubMed DOI PMC

Breitkreutz I, Raab MS, Vallet S, Hideshima T, Raje N, Mitsiades C, Chauhan D, Okawa Y, Munshi NC, Richardson PG et al., Lenalidomide inhibits osteoclastogenesis, survival factors and bone-remodeling markers in multiple myeloma. Leukemia 2008; 22(10):1925-32; PMID:18596740; http://dx.doi.org/ 10.1038/leu.2008.174 PubMed DOI

Ramsay AG, Johnson AJ, Lee AM, Gorgün G, Le Dieu R, Blum W, Byrd JC, Gribben JG. Chronic lymphocytic leukemia T cells show impaired immunological synapse formation that can be reversed with an immunomodulating drug. J Clin Invest 2008; 118(7):2427-37; PMID:18551193 PubMed PMC

Ramsay AG, Clear AJ, Fatah R, Gribben JG. Multiple inhibitory ligands induce impaired T-cell immunologic synapse function in chronic lymphocytic leukemia that can be blocked with lenalidomide: establishing a reversible immune evasion mechanism in human cancer. Blood 2012; 120 (7):1412-21; PMID:22547582; http://dx.doi.org/ 10.1182/blood-2012-02-411678 PubMed DOI PMC

Galustian C, Meyer B, Labarthe MC, Dredge K, Klaschka D, Henry J, Todryk S, Chen R, Muller G, Stirling D et al.. The anti-cancer agents lenalidomide and pomalidomide inhibit the proliferation and function of T regulatory cells. Cancer Immunol Immunother 2009; 58(7):1033-45; PMID:19009291; http://dx.doi.org/ 10.1007/s00262-008-0620-4 PubMed DOI PMC

Kneppers E, van der Holt B, Kersten MJ, Zweegman S, Meijer E, Huls G, Cornelissen JJ, Janssen JJ, Huisman C, Cornelisse PB et al., Lenalidomide maintenance after nonmyeloablative allogeneic stem cell transplantation in multiple myeloma is not feasible: results of the HOVON 76 Trial.Blood 2011; 118(9):2413-9; PMID:21690556; http://dx.doi.org/23765229 10.1182/blood-2011-04-348292 PubMed DOI

Sakamaki I, Kwak LW, Cha SC, Yi Q, Lerman B, Chen J, Surapaneni S, Bateman S, Qin H. Lenalidomide enhances the protective effect of a therapeutic vaccine and reverses immune suppression in mice bearing established lymphomas. Leukemia 2014; 28(2):329-37; PMID:23765229; http://dx.doi.org/ 10.1038/leu.2013.177 PubMed DOI PMC

Haslett PA, Hanekom WA, Muller G, Kaplan G. Thalidomide and a thalidomide analogue drug costimulate virus-specific CD8+ T cells in vitro. J Infect Dis 2003; 187 (6):946-55; PMID:12660941; http://dx.doi.org/ 10.1086/368126 PubMed DOI

Franklin RA, Tordai A, Patel H, Gardner AM, Johnson GL, Gelfand EW. Ligation of the T cell receptor complex results in activation of the Ras/Raf-1/MEK/MAPK cascade in human T lymphocytes. J Clin Invest 1994; 93(5):2134-40; PMID:8182145; http://dx.doi.org/ 10.1172/JCI117209 PubMed DOI PMC

Marchingo JM, Kan A, Sutherland RM, Duffy KR, Wellard CJ, Belz GT, Lew AM, Dowling MR, Heinzel S, Hodgkin PD. T cell signaling. Antigen affinity, costimulation, and cytokine inputs sum linearly to amplify T cell expansion. Science 2014; 346(6213):1123-7; PMID:25430770; http://dx.doi.org/ 10.1126/science.1260044 PubMed DOI

Till BG, Jensen MC, Wang J, Qian X, Gopal AK, Maloney DG, Lindgren CG, Lin Y et al., CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results. Blood 2012; 119(17):3940-50; PMID:22308288; http://dx.doi.org/ 10.1182/blood-2011-10-387969 PubMed DOI PMC

Han EQ, Li XL, Wang CR, Li TF, Han SY. Chimeric antigen receptor-engineered T cells for cancer immunotherapy: progress and challenges. J Hematol Oncol 2013; 6:47; PMID:23829929; http://dx.doi.org/ 10.1186/1756-8722-6-47 PubMed DOI PMC