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A novel chimeric antigen receptor containing a JAK–STAT signaling domain mediates superior antitumor effects

Nature Medicine volume 24, pages 352–359 (2018)Cite this article

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Abstract

The adoptive transfer of T cells engineered with a chimeric antigen receptor (CAR) (hereafter referred to as CAR-T cells) specific for the B lymphocyte antigen CD19 has shown impressive clinical responses in patients with refractory B cell malignancies1,2,3,4,5,6,7. However, the therapeutic effects of CAR-T cells that target other malignancies have not yet resulted in significant clinical benefit8,9,10,11. Although inefficient tumor trafficking and various immunosuppressive mechanisms can impede CAR-T cell effector responses, the signals delivered by the current CAR constructs may still be insufficient to fully activate antitumor T cell functions. Optimal T cell activation and proliferation requires multiple signals, including T cell receptor (TCR) engagement (signal 1), co-stimulation (signal 2) and cytokine engagement (signal 3)12. However, CAR constructs currently being tested in the clinic contain a CD3z (TCR signaling) domain and co-stimulatory domain(s) but not a domain that transmits signal 3 (refs. 13, 14, 15, 16, 17, 18). Here we have developed a novel CAR construct capable of inducing cytokine signaling after antigen stimulation. This new-generation CD19 CAR encodes a truncated cytoplasmic domain from the interleukin (IL)-2 receptor β-chain (IL-2Rβ) and a STAT3-binding tyrosine-X-X-glutamine (YXXQ) motif, together with the TCR signaling (CD3z) and co-stimulatory (CD28) domains (hereafter referred to as 28-ΔIL2RB-z(YXXQ)). The 28-ΔIL2RB-z(YXXQ) CAR-T cells showed antigen-dependent activation of the JAK kinase and of the STAT3 and STAT5 transcription factors signaling pathways, which promoted their proliferation and prevented terminal differentiation in vitro. The 28-ΔIL2RB-z(YXXQ) CAR-T cells demonstrated superior in vivo persistence and antitumor effects in models of liquid and solid tumors as compared with CAR-T cells expressing a CD28 or 4-1BB co-stimulatory domain alone. Taken together, these results suggest that our new-generation CAR has the potential to demonstrate superior antitumor effects with minimal toxicity in the clinic and that clinical translation of this novel CAR is warranted.

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Figure 1: Generation of the CD19-specific chimeric antigen receptor constructs to induce JAK–STAT pathway activation.
Figure 2: The 28-ΔIL2RB-z(YXXQ) CAR-T cells show a superior proliferative capacity and maintain less differentiated memory T cell phenotypes after antigen stimulation.
Figure 3: The 28-ΔIL2RB-z(YXXQ) CAR-T cells have unique gene expression profiles and show potent cytotoxic activity after repetitive antigen stimulations.
Figure 4: T cells transduced with the 28-ΔIL2RB-z(YXXQ) CAR have superior antitumor effects in vivo.

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Acknowledgements

This work was supported by CIHR Project Grant 362860 (N.H.), Ontario Institute for Cancer Research Clinical Investigator Award IA-039 (N.H.), BioCanRX Catalyst Program grant FY17CAT7 (N.H.), the Princess Margaret Cancer Foundation (M.O.B. and N.H.), a Japan Society for the Promotion of Science Postdoctoral Fellowship for Overseas Researchers (Y.K.), a Guglietti Fellowship Award (Y.K.), a Canadian Institutes of Health Research Canada Graduate Scholarship (T.G.), the Province of Ontario (T.G. and M.A.) and a Natural Sciences and Engineering Research Council of Canada Postgraduate Scholarship (T.G.). This study was partly sponsored by Takara Bio, Inc.

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Authors and Affiliations

  1. Tumor Immunotherapy Program, Campbell Family Institute for Breast Cancer Research, Campbell Family Cancer Research Institute, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada

    Yuki Kagoya, Shinya Tanaka, Tingxi Guo, Mark Anczurowski, Chung-Hsi Wang, Kayoko Saso, Marcus O Butler & Naoto Hirano

  2. Takara Bio, Inc., Kusatsu, Japan

    Shinya Tanaka

  3. Department of Immunology, University of Toronto, Toronto, Ontario, Canada

    Tingxi Guo, Mark Anczurowski, Chung-Hsi Wang, Marcus O Butler & Naoto Hirano

  4. Department of Medicine, University of Toronto, Toronto, Ontario, Canada

    Marcus O Butler

  5. Department of Hematology–Oncology, Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada

    Mark D Minden

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Contributions

Y.K. and N.H. designed the project; Y.K., S.T., T.G., M.A., C.-H.W. and K.S. performed the experiments; M.D.M. and M.O.B. provided critical human samples and contributed to the writing of the manuscript; and Y.K. and N.H. analyzed the results and wrote the manuscript.

Corresponding author

Correspondence to Naoto Hirano.

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Competing interests

S.T. is an employee of Takara Bio, Inc. This study was partly sponsored by Takara Bio, Inc. The University Health Network has filed a patent application related to this study on which N.H., Y.K. and S.T. are named as inventors.

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Kagoya, Y., Tanaka, S., Guo, T. et al. A novel chimeric antigen receptor containing a JAK–STAT signaling domain mediates superior antitumor effects. Nat Med 24, 352–359 (2018). https://doi.org/10.1038/nm.4478

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