De novo-designed transmembrane domains tune engineered receptor functions

• Main Authors: Call, M.E (Author), Call, M.J (Author), Chandler, N.J (Author), Cross, R.S (Author), Davey, A.S (Author), Elazar, A. (Author), Fleishman, S.J (Author), Jenkins, M.R (Author), Nguyen, J.V (Author), Trenker, R. (Author), Weinstein, J.Y (Author)
• Format: Article
• Language: English
• Published: NLM (Medline) 2022
• Subjects: CAR T cell; chimeric antigen receptor; de novo design; E. coli; immunology; immunotherapy; inflammation; membrane protein; molecular biophysics; mouse; Rosetta; structural biology; transmembrane
• Online Access: View Fulltext in Publisher

 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. © 2022, Elazar et al.