Co-opting signalling molecules enables logic-gated control of CAR T cells

• Published in: Nature (London) Vol. 615; no. 7952; pp. 507 - 516
• Main Authors: Tousley, Aidan M., Rotiroti, Maria Caterina, Labanieh, Louai, Rysavy, Lea Wenting, Kim, Won-Ju, Lareau, Caleb, Sotillo, Elena, Weber, Evan W., Rietberg, Skyler P., Dalton, Guillermo Nicolas, Yin, Yajie, Klysz, Dorota, Xu, Peng, de la Serna, Eva L., Dunn, Alexander R., Satpathy, Ansuman T., Mackall, Crystal L., Majzner, Robbie G.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.03.2023; Nature Publishing Group
• Subjects: 13/31; 42/41; 59/5; 631/61/338/552; 631/67/1059/2325; 631/80/86; 64/60; 692/308/575; 82/1; 82/80; 96/106; 96/21; 96/44; Antigens; Biocompatibility; Boolean; Cell Engineering - methods; Channel gating; Chimeric antigen receptors; Cytokines; Humanities and Social Sciences; Humans; Immunotherapy, Adoptive - adverse effects; Intracellular; Intracellular signalling; Kinases; Leukemia, B-Cell; Logic; Lymphocytes; Lymphocytes T; Lymphoma, B-Cell; Malignancy; multidisciplinary; Neoplasms - immunology; Neoplasms - metabolism; Neoplasms - therapy; Pharmacology; Receptors; Receptors, Antigen, T-Cell - immunology; Receptors, Antigen, T-Cell - metabolism; Receptors, Chimeric Antigen - immunology; Receptors, Chimeric Antigen - metabolism; Science; Science (multidisciplinary); Signal Transduction; Signaling; SLP-76 protein; Solid tumors; T cell receptors; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; Toxicity; Tumor necrosis factor-TNF; Tumors; ZAP-70 protein
• ISSN: 0028-0836; 1476-4687; 1476-4687
• DOI: 10.1038/s41586-023-05778-2

Although chimeric antigen receptor (CAR) T cells have altered the treatment landscape for B cell malignancies, the risk of on-target, off-tumour toxicity has hampered their development for solid tumours because most target antigens are shared with normal cells 1 , 2 . Researchers have attempted to apply Boolean-logic gating to CAR T cells to prevent toxicity 3 – 5 ; however, a truly safe and effective logic-gated CAR has remained elusive 6 . Here we describe an approach to CAR engineering in which we replace traditional CD3ζ domains with intracellular proximal T cell signalling molecules. We show that certain proximal signalling CARs, such as a ZAP-70 CAR, can activate T cells and eradicate tumours in vivo while bypassing upstream signalling proteins, including CD3ζ. The primary role of ZAP-70 is to phosphorylate LAT and SLP-76, which form a scaffold for signal propagation. We exploited the cooperative role of LAT and SLP-76 to engineer logic-gated intracellular network (LINK) CAR, a rapid and reversible Boolean-logic AND-gated CAR T cell platform that outperforms other systems in both efficacy and prevention of on-target, off-tumour toxicity. LINK CAR will expand the range of molecules that can be targeted with CAR T cells, and will enable these powerful therapeutic agents to be used for solid tumours and diverse diseases such as autoimmunity 7 and fibrosis 8 . In addition, this work shows that the internal signalling machinery of cells can be repurposed into surface receptors, which could open new avenues for cellular engineering. Logic gating is used to develop a CAR T cell platform that is highly specific and allows the activity of T cells to be restricted to the encounter of two antigens, thus reducing on-target, off-tumour toxicity.