Control of the antitumour activity and specificity of CAR T cells via organic adapters covalently tethering the CAR to tumour cells

Alexey V Stepanov; Jia Xie; Qiaoqiao Zhu; Zuyuan Shen; Wenji Su; Letian Kuai; Richard Soll; Christoph Rader; Geramie Shaver; Lacey Douthit; Ding Zhang; Roman Kalinin; Xiang Fu; Yingying Zhao; Tian Qin; Phil S Baran; Alexander G Gabibov; David Bushnell; Dario Neri; Roger D Kornberg; Richard A Lerner

doi:10.1038/s41551-023-01102-5

Control of the antitumour activity and specificity of CAR T cells via organic adapters covalently tethering the CAR to tumour cells

Nat Biomed Eng. 2023 Oct 5. doi: 10.1038/s41551-023-01102-5. Online ahead of print.

Authors

Alexey V Stepanov^#¹, Jia Xie^#², Qiaoqiao Zhu³, Zuyuan Shen³, Wenji Su³, Letian Kuai³, Richard Soll³, Christoph Rader⁴, Geramie Shaver², Lacey Douthit², Ding Zhang², Roman Kalinin⁵, Xiang Fu², Yingying Zhao², Tian Qin⁶, Phil S Baran², Alexander G Gabibov⁵, David Bushnell⁷, Dario Neri⁸, Roger D Kornberg⁹, Richard A Lerner²

Affiliations

¹ Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA. stepanov@scripps.edu.
² Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA.
³ WuXi AppTec Co., Ltd, Shanghai, China.
⁴ Department of Immunology and Microbiology, UF Scripps Biomedical Research, University of Florida, Jupiter, FL, USA.
⁵ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.
⁶ The University of Texas Southwestern Medical Center, Dallas, TX, USA.
⁷ Structural Biology, School of Medicine, Stanford University, Stanford, CA, USA.
⁸ Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
⁹ Structural Biology, School of Medicine, Stanford University, Stanford, CA, USA. kornberg@stanford.edu.

^# Contributed equally.

PMID: 37798444
DOI: 10.1038/s41551-023-01102-5

Abstract

On-target off-tumour toxicity limits the anticancer applicability of chimaeric antigen receptor (CAR) T cells. Here we show that the tumour-targeting specificity and activity of T cells with a CAR consisting of an antibody with a lysine residue that catalytically forms a reversible covalent bond with a 1,3-diketone hapten can be regulated by the concentration of a small-molecule adapter. This adapter selectively binds to the hapten and to a chosen tumour antigen via a small-molecule binder identified via a DNA-encoded library. The adapter therefore controls the formation of a covalent bond between the catalytic antibody and the hapten, as well as the tethering of the CAR T cells to the tumour cells, and hence the cytotoxicity and specificity of the cytotoxic T cells, as we show in vitro and in mice with prostate cancer xenografts. Such small-molecule switches of T-cell cytotoxicity and specificity via an antigen-independent 'universal' CAR may enhance the control and safety profile of CAR-based cellular immunotherapies.