De novo-designed transmembrane domains tune engineered receptor functions

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...

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Bibliographic Details
Published in:eLife 2022-05, Vol.11
Main Authors: Elazar, Assaf, Chandler, Nicholas J, Davey, Ashleigh S, Weinstein, Jonathan Y, Nguyen, Julie V, Trenker, Raphael, Cross, Ryan S, Jenkins, Misty R, Call, Melissa J, Call, Matthew E, Fleishman, Sarel J
Format: Article
Language:English
Subjects:
Amino acids
Animals
Antigens
Antitumor activity
CAR T cell
CD28 antigen
CD28 Antigens - metabolism
chimeric antigen receptor
Chimeric antigen receptors
Cytokines
Cytokines - metabolism
de novo design
Design
Energy
Immunology and Inflammation
Immunotherapy
Interfaces
Lymphocytes
Lymphocytes T
membrane protein
Membrane proteins
Mice
Monomers
Mutation
Peptides
Protein Domains
Proteins
Receptors, Antigen, T-Cell - metabolism
Receptors, Chimeric Antigen - metabolism
Structural Biology and Molecular Biophysics
T cell receptors
T-Lymphocytes
transmembrane
Transmembrane domains
Xenograft Model Antitumor Assays
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Summary: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.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.75660