De novo-designed transmembrane domains tune engineered receptor functions

Assaf Elazar, Nicholas J Chandler, Ashleigh S Davey, Jonathan Y Weinstein, Julie V Nguyen, Raphael Trenker, Ryan S Cross, Misty R Jenkins, Melissa J Call, Matthew E Call, Sarel J Fleishman

Research output: Contribution to journalArticlepeer-review

Abstract

De novo-designed receptor transmembrane domains (TMDs) present opportunities for precise control of cellular receptor functions. We developed a de novo design strategy for generating programmed membrane proteins (proMPs): single-pass α-helical TMDs that self-assemble through computationally defined and crystallographically validated interfaces. We used these proMPs to program specific oligomeric interactions into a chimeric antigen receptor (CAR) that we expressed in mouse primary T cells and found that both in vitro CAR T cell cytokine release and in vivo antitumor activity scaled linearly with the oligomeric state encoded by the receptor TMD, from monomers up to tetramers. All programmed CARs stimulated substantially lower T cell cytokine release relative to the commonly used CD28 TMD, which we show elevated cytokine release through lateral recruitment of the endogenous T cell costimulatory receptor CD28. Precise design using orthogonal and modular TMDs thus provides a new way to program receptor structure and predictably tune activity for basic or applied synthetic biology.
Original languageEnglish
Article numbere75660
Number of pages29
JournaleLife
Volume11
DOIs
StatePublished - 4 May 2022

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