Abstract
Chimeric antigen receptor (CAR)-expressing T cells targeting surface-bound tumor antigens have yielded promising clinical outcomes, with two CD19 CAR-T cell therapies recently receiving FDA approval for the treatment of B-cell malignancies. The adoption of CARs for the recognition of soluble ligands, a distinct class of biomarkers in physiology and disease, could considerably broaden the utility of CARs in disease treatment. In this study, we demonstrate that CAR-T cells can be engineered to respond robustly to diverse soluble ligands, including the CD19 ectodomain, GFP variants, and transforming growth factor beta (TGF-β). We additionally show that CAR signaling in response to soluble ligands relies on ligand-mediated CAR dimerization and that CAR responsiveness to soluble ligands can be fine-tuned by adjusting the mechanical coupling between the CAR's ligand-binding and signaling domains. Our results support a role for mechanotransduction in CAR signaling and demonstrate an approach for systematically engineering immune-cell responses to soluble, extracellular ligands.
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Acknowledgements
This work was supported by the National Institutes of Health (DP5OD012133, grant to Y.Y.C.; F30CA183528, fellowship to Z.L.C.). We thank M. Jensen (Seattle Children's Research Institute), S. Forman (City of Hope National Medical Center), D. Kohn (University of California, Los Angeles), X. Lin (University of Texas MD Anderson Cancer Center), A. Weiss (University of California, San Francisco), and T. Yeates (University of California, Los Angeles) for materials used in this work. We also thank Y. Choi, H. Ho, and R. Smolkin for assistance and support in the lab.
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Z.L.C. and Y.Y.C. designed the project, participated in data analysis throughout, and wrote the manuscript. Z.L.C., M.H.L., and Y.Y.C. edited and revised the manuscript. Z.L.C. developed the TGF-β CAR system and performed and analyzed microscopy, western blot, and computational modeling experiments. Z.L.C., M.H.L., and U.T. performed and analyzed TGF-β CAR flow cytometry, cytokine production, and cell expansion experiments. X.C. performed the CD19 CAR experiments. Z.L.C. and N.J.B. developed and tested the GFP CAR system.
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Z.L.C. and Y.Y.C. declare competing financial interests in the form of a pending patent application whose value may be affected by the publication of this manuscript.
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Lack of bystander cell lysis by TGF-β CAR-T cells (XLSX 38 kb)
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Chang, Z., Lorenzini, M., Chen, X. et al. Rewiring T-cell responses to soluble factors with chimeric antigen receptors. Nat Chem Biol 14, 317–324 (2018). https://doi.org/10.1038/nchembio.2565
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DOI: https://doi.org/10.1038/nchembio.2565
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