Nature Communications (May 2023)

Post-translational covalent assembly of CAR and synNotch receptors for programmable antigen targeting

  • Elisa Ruffo,
  • Adam A. Butchy,
  • Yaniv Tivon,
  • Victor So,
  • Michael Kvorjak,
  • Avani Parikh,
  • Eric L. Adams,
  • Natasa Miskov-Zivanov,
  • Olivera J. Finn,
  • Alexander Deiters,
  • Jason Lohmueller

DOI
https://doi.org/10.1038/s41467-023-37863-5
Journal volume & issue
Vol. 14, no. 1
pp. 1 – 16

Abstract

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Abstract Chimeric antigen receptors (CARs) and synthetic Notch (synNotch) receptors are engineered cell-surface receptors that sense a target antigen and respond by activating T cell receptor signaling or a customized gene program, respectively. Here, to expand the targeting capabilities of these receptors, we develop “universal” receptor systems for which receptor specificity can be directed post-translationally via covalent attachment of a co-administered antibody bearing a benzylguanine (BG) motif. A SNAPtag self-labeling enzyme is genetically fused to the receptor and reacts with BG-conjugated antibodies for covalent assembly, programming antigen recognition. We demonstrate that activation of SNAP-CAR and SNAP-synNotch receptors can be successfully targeted by clinically relevant BG-conjugated antibodies, including anti-tumor activity of SNAP-CAR T cells in vivo in a human tumor xenograft mouse model. Finally, we develop a mathematical model to better define the parameters affecting universal receptor signaling. SNAP receptors provide a powerful strategy to post-translationally reprogram the targeting specificity of engineered cells.