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Immunotherapy

Cytokine IL-36γ improves CAR T-cell functionality and induces endogenous antitumor response

Leukemia volume 35, pages 506–521 (2021)Cite this article

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Abstract

Chimeric antigen receptor (CAR) T-cell therapy has shown remarkable responses in B-cell malignancies. However, many patients suffer from limited response and tumor relapse due to lack of persisting CAR T cells and immune escape. These clinical challenges have compromised the long-term efficacy of CAR T-cell therapy and call for the development of novel CAR designs. We demonstrated that CAR T cells secreting a cytokine interleukin-36γ (IL-36γ) showed significantly improved CAR T-cell expansion and persistence, and resulted in superior tumor eradication compared with conventional CAR T cells. The enhanced cellular function by IL-36γ was mediated through an autocrine manner. In addition, activation of endogenous antigen-presenting cells (APCs) and T cells by IL-36γ aided the formation of a secondary antitumor response, which delayed the progression of antigen-negative tumor challenge. Together, our data provide preclinical evidence to support the translation of this design for an improved CAR T-cell-mediated antitumor response.

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Fig. 1: CAR T-cell construct validation and in vitro characterization.
Fig. 2: IL-36γ-secreting CAR T cell exhibited superior antitumor efficacy with and without preconditioning.
Fig. 3: IL-36γ enhances CAR T-cell proliferation and cytokine secretion ex vivo.
Fig. 4: Antitumor efficacy requires the CD8+ CAR T-cell subset, and is mediated by autocrine IL-36 signaling via the Myd88 pathway.
Fig. 5: IL-36γ activates myeloid cells and exposure to IL-36γ-secreting CAR T cells promotes tumor recognition by endogenous T cells.
Fig. 6: IL-36γ-secreting CAR T-cell enhances tumor eradication in a xenograft model.

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References

  1. Brentjens RJ, Davila ML, Riviere I, Park J, Wang X, Cowell LG, et al. CD19-targeted T cells rapidly induce molecular remissions in adults with chemotherapy-refractory acute lymphoblastic leukemia. Sci Transl Med. 2013;5:177ra38.

    Article  Google Scholar 

  2. 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. 2015;33:540–9.

    Article  CAS  Google Scholar 

  3. 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;365:725–33.

    Article  CAS  Google Scholar 

  4. Turtle CJ, Hanafi LA, Berger C, Hudecek M, Pender B, Robinson E, et al. Immunotherapy of non-Hodgkin’s lymphoma with a defined ratio of CD8+ and CD4+ CD19-specific chimeric antigen receptor-modified T cells. Sci Transl Med. 2016;8:355ra116.

    Article  Google Scholar 

  5. Maude SL, Laetsch TW, Buechner J, Rives S, Boyer M, Bittencourt H, et al. Tisagenlecleucel in children and young adults with B-cell lymphoblastic leukemia. N Engl J Med. 2018;378:439–48.

    Article  CAS  Google Scholar 

  6. Park JH, Riviere I, Gonen M, Wang X, Senechal B, Curran KJ, et al. Long-term follow-up of CD19 CAR therapy in acute lymphoblastic leukemia. N Engl J Med. 2018;378:449–59.

    Article  CAS  Google Scholar 

  7. Majzner RG, Mackall CL. Tumor antigen escape from CAR T-cell therapy. Cancer Discov. 2018;8:1219–26.

    Article  CAS  Google Scholar 

  8. Fraietta JA, Lacey SF, Orlando EJ, Pruteanu-Malinici I, Gohil M, Lundh S, et al. Determinants of response and resistance to CD19 chimeric antigen receptor (CAR) T cell therapy of chronic lymphocytic leukemia. Nat Med. 2018;24:563–71.

    Article  CAS  Google Scholar 

  9. Rossi J, Paczkowski P, Shen YW, Morse K, Flynn B, Kaiser A, et al. Preinfusion polyfunctional anti-CD19 chimeric antigen receptor T cells are associated with clinical outcomes in NHL. Blood. 2018;132:804–14.

    Article  CAS  Google Scholar 

  10. Feucht J, Sun J, Eyquem J, Ho YJ, Zhao Z, Leibold J, et al. Calibration of CAR activation potential directs alternative T cell fates and therapeutic potency. Nat Med. 2019;25:82–8.

    Article  CAS  Google Scholar 

  11. Zhao Z, Condomines M, van der Stegen SJC, Perna F, Kloss CC, Gunset G, et al. Structural design of engineered costimulation determines tumor rejection kinetics and persistence of CAR T cells. Cancer Cell. 2015;28:415–28.

    Article  CAS  Google Scholar 

  12. Kuhn NF, Purdon TJ, van Leeuwen DG, Lopez AV, Curran KJ, Daniyan AF, et al. CD40 ligand-modified chimeric antigen receptor T cells enhance antitumor function by eliciting an endogenous antitumor response. Cancer Cell. 2019;35:473–88.e6.

    Article  CAS  Google Scholar 

  13. Pegram HJ, Lee JC, Hayman EG, Imperato GH, Tedder TF, Sadelain M, et al. Tumor-targeted T cells modified to secrete IL-12 eradicate systemic tumors without need for prior conditioning. Blood. 2012;119:4133–41.

    Article  CAS  Google Scholar 

  14. Rafiq S, Yeku OO, Jackson HJ, Purdon TJ, van Leeuwen DG, Drakes DJ, et al. Targeted delivery of a PD-1-blocking scFv by CAR-T cells enhances anti-tumor efficacy in vivo. Nat Biotechnol. 2018;36:847–56.

    Article  CAS  Google Scholar 

  15. Yeku OO, Purdon TJ, Koneru M, Spriggs D, Brentjens RJ. Armored CAR T cells enhance antitumor efficacy and overcome the tumor microenvironment. Sci Rep. 2017;7:10541.

    Article  Google Scholar 

  16. Avanzi MP, Yeku O, Li X, Wijewarnasuriya DP, van Leeuwen DG, Cheung K, et al. Engineered tumor-targeted T cells mediate enhanced anti-tumor efficacy both directly and through activation of the endogenous immune system. Cell Rep. 2018;23:2130–41.

    Article  CAS  Google Scholar 

  17. Chmielewski M, Abken H. CAR T cells releasing IL-18 convert to T-Bet(high) FoxO1(low) effectors that exhibit augmented activity against advanced solid tumors. Cell Rep. 2017;21:3205–19.

    Article  CAS  Google Scholar 

  18. Hu B, Ren J, Luo Y, Keith B, Young RM, Scholler J, et al. Augmentation of antitumor immunity by human and mouse CAR T cells secreting IL-18. Cell Rep. 2017;20:3025–33.

    Article  CAS  Google Scholar 

  19. Gabay C, Towne JE. Regulation and function of interleukin-36 cytokines in homeostasis and pathological conditions. J Leukoc Biol. 2015;97:645–52.

    Article  CAS  Google Scholar 

  20. Gresnigt MS, van de Veerdonk FL. Biology of IL-36 cytokines and their role in disease. Semin Immunol. 2013;25:458–65.

    Article  CAS  Google Scholar 

  21. Vigne S, Palmer G, Martin P, Lamacchia C, Strebel D, Rodriguez E, et al. IL-36 signaling amplifies Th1 responses by enhancing proliferation and Th1 polarization of naive CD4+ T cells. Blood. 2012;120:3478–87.

    Article  CAS  Google Scholar 

  22. Wang X, Zhao X, Feng C, Weinstein A, Xia R, Wen W, et al. IL-36gamma transforms the tumor microenvironment and promotes type 1 lymphocyte-mediated antitumor immune responses. Cancer Cell. 2015;28:296–306.

    Article  CAS  Google Scholar 

  23. Weinstein AM, Chen L, Brzana EA, Patil PR, Taylor JL, Fabian KL, et al. Tbet and IL-36gamma cooperate in therapeutic DC-mediated promotion of ectopic lymphoid organogenesis in the tumor microenvironment. Oncoimmunology. 2017;6:e1322238.

    Article  Google Scholar 

  24. Lee J, Sadelain M, Brentjens R. Retroviral transduction of murine primary T lymphocytes. Methods Mol Biol. 2009;506:83–96.

    Article  CAS  Google Scholar 

  25. Gade TP, Hassen W, Santos E, Gunset G, Saudemont A, Gong MC, et al. Targeted elimination of prostate cancer by genetically directed human T lymphocytes. Cancer Res. 2005;65:9080–8.

    Article  CAS  Google Scholar 

  26. Helft J, Bottcher J, Chakravarty P, Zelenay S, Huotari J, Schraml BU, et al. GM-CSF mouse bone marrow cultures comprise a heterogeneous population of CD11c(+)MHCII(+) macrophages and dendritic cells. Immunity. 2015;42:1197–211.

    Article  CAS  Google Scholar 

  27. Philip M, Fairchild L, Sun L, Horste EL, Camara S, Shakiba M, et al. Chromatin states define tumour-specific T cell dysfunction and reprogramming. Nature. 2017;545:452–6.

    Article  CAS  Google Scholar 

  28. Sommermeyer D, Hudecek M, Kosasih PL, Gogishvili T, Maloney DG, Turtle CJ, et al. Chimeric antigen receptor-modified T cells derived from defined CD8+ and CD4+ subsets confer superior antitumor reactivity in vivo. Leukemia. 2016;30:492–500.

    Article  CAS  Google Scholar 

  29. Radvanyi LG, Bernatchez C, Zhang M, Fox PS, Miller P, Chacon J, et al. Specific lymphocyte subsets predict response to adoptive cell therapy using expanded autologous tumor-infiltrating lymphocytes in metastatic melanoma patients. Clin Cancer Res. 2012;18:6758–70.

    Article  CAS  Google Scholar 

  30. Vigne S, Palmer G, Lamacchia C, Martin P, Talabot-Ayer D, Rodriguez E, et al. IL-36R ligands are potent regulators of dendritic and T cells. Blood. 2011;118:5813–23.

    Article  CAS  Google Scholar 

  31. Foster AM, Baliwag J, Chen CS, Guzman AM, Stoll SW, Gudjonsson JE, et al. IL-36 promotes myeloid cell infiltration, activation, and inflammatory activity in skin. J Immunol. 2014;192:6053–61.

    Article  CAS  Google Scholar 

  32. Neelapu SS, Locke FL, Bartlett NL, Lekakis LJ, Miklos DB, Jacobson CA, et al. Axicabtagene ciloleucel CAR T-cell therapy in refractory large B-cell lymphoma. N Engl J Med. 2017;377:2531–44.

    Article  CAS  Google Scholar 

  33. Cohen ES, Scott IC, Majithiya JB, Rapley L, Kemp BP, England E, et al. Oxidation of the alarmin IL-33 regulates ST2-dependent inflammation. Nat Commun. 2015;6:8327.

    Article  CAS  Google Scholar 

  34. Mata M, Gerken C, Nguyen P, Krenciute G, Spencer DM, Gottschalk S. Inducible activation of MyD88 and CD40 in CAR T cells results in controllable and potent antitumor activity in preclinical solid tumor models. Cancer Discov. 2017;7:1306–19.

    Article  CAS  Google Scholar 

  35. Collinson-Pautz MR, Chang WC, Lu A, Khalil M, Crisostomo JW, Lin PY, et al. Constitutively active MyD88/CD40 costimulation enhances expansion and efficacy of chimeric antigen receptor T cells targeting hematological malignancies. Leukemia. 2019;33:2195–207.

    Article  CAS  Google Scholar 

  36. Giavridis T, van der Stegen SJC, Eyquem J, Hamieh M, Piersigilli A, Sadelain M. CAR T cell-induced cytokine release syndrome is mediated by macrophages and abated by IL-1 blockade. Nat Med. 2018;24:731–8.

    Article  CAS  Google Scholar 

  37. Norelli M, Camisa B, Barbiera G, Falcone L, Purevdorj A, Genua M, et al. Monocyte-derived IL-1 and IL-6 are differentially required for cytokine-release syndrome and neurotoxicity due to CAR T cells. Nat Med. 2018;24:739–48.

    Article  CAS  Google Scholar 

  38. Chmielewski M, Kopecky C, Hombach AA, Abken H. IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression. Cancer Res. 2011;71:5697–706.

    Article  CAS  Google Scholar 

  39. Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH, et al. Engineering T cells with customized therapeutic response programs using synthetic notch receptors. Cell. 2016;167:419–32.e16.

  40. Roybal KT, Rupp LJ, Morsut L, Walker WJ, McNally KA, Park JS, et al. Precision tumor recognition by T cells with combinatorial antigen-sensing circuits. Cell. 2016;164:770–9.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Antoinette Rookard and the Antitumor Assessment Core Facility of Memorial Sloan Kettering Cancer Center (MSK) for their assistance with animal breeding and in vivo experiments. We thank Amy Plofker for her editing on the manuscript. We also acknowledge Amgen for providing IL-36R−/− mice for this study.

Funding

The study was financially supported by US National Institutes of Health grants P30 CA008748 (Cancer Center Support Grant), 5 P01 CA190174-03, and 5 P50 CA192937-02 (RJB), The Annual Terry Fox Run for Cancer Research organized by the Canada Club of New York (RJB), Kate’s Team (RJB), Carson Family Charitable Trust (RJB), Mr William H. Goodwin and Mrs Alice Goodwin and the Commonwealth Foundation for Cancer Research and the Experimental Therapeutics Center of MSK (Innovations in the structures, functions, and targets of monoclonal antibody-based drugs for cancer) (RJB).

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Author notes

  1. These authors contributed equally: Xinghuo Li, Anthony F. Daniyan

Authors and Affiliations

  1. Weill Cornell Graduate School of Medical Sciences, New York, NY, 10065, USA

    Xinghuo Li & Renier J. Brentjens

  2. Department of Medicine, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA

    Xinghuo Li, Anthony F. Daniyan, Andrea V. Lopez, Terence J. Purdon & Renier J. Brentjens

  3. Molecular Pharmacology & Chemistry Program, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA

    Renier J. Brentjens

  4. Center for Cell Engineering, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA

    Renier J. Brentjens

  5. Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA

    Renier J. Brentjens

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  1. Xinghuo Li
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Contributions

XL and AFD designed the study; XL, AFD, AVL, and TJP conducted the experiments; XL performed data analysis; RJB supervised the project; and XL, AFD, and RJB drafted the manuscript.

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Correspondence to Renier J. Brentjens.

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Conflict of interest

AFD and RJB have submitted a patent related to this work. RJB is a co-founder of and receives royalties from Juno Therapeutics/Celgene. The other authors declare that they have no conflict of interest.

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Li, X., Daniyan, A.F., Lopez, A.V. et al. Cytokine IL-36γ improves CAR T-cell functionality and induces endogenous antitumor response. Leukemia 35, 506–521 (2021). https://doi.org/10.1038/s41375-020-0874-1

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