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bioRxiv · 10.1101/2020.01.17.909895

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

Lohmueller, J.·Butchy, A. A.·Tivon, Y.·Kvorjak, M.·Miskov-Zivanov, N.·Deiters, A.·Finn, O. J.
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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. To expand the targeting capabilities of these receptors, we have developed switchable adaptor receptor systems for which receptor specificity can be directed post-translationally via covalent attachment of a co-administered antibody. Instead of directly targeting an antigen, our receptors contain the SNAPtag self-labeling enzyme, which reacts with benzylguanine (BG)-conjugated antibodies to assemble covalently-associated antigen receptors. We demonstrate that activation of SNAP-CAR and SNAP-synNotch receptors and their downstream effector functions can be successfully targeted by several clinically-relevant BG-conjugated antibodies in an antigen-specific and antibody dose-dependent manner. To better define parameters affecting receptor signaling, we developed a mathematical model of switchable receptor systems. SNAP receptors provide a powerful new strategy to post-translationally reprogram the targeting specificity of engineered cells.

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Lohmueller, J., Butchy, A. A., Tivon, Y., Kvorjak, M., Miskov-Zivanov, N., Deiters, A., Finn, O. J.. 2020-01-17. Post-translational covalent assembly of CAR and synNotch receptors for programmable antigen targeting. https://doi.org/10.1101/2020.01.17.909895

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