Giordano-Attianese, GretaGainza, PabloGray-Gaillard, EliseCribioli, ElisabettaShui, SailanKim, SeonghoonKwak, Mi-JeongVollers, SabrinaOsorio, Angel De Jesus CorriaReichenbach, PatrickBonet, JaumeOh, Byung-HaIrving, MelitaCoukos, GeorgeCorreia, Bruno E.2020-03-032020-09-012020-03-032020-02-0310.1038/s41587-019-0403-9https://infoscience.epfl.ch/handle/20.500.14299/166849WOS:000510822200003The activity of CAR-T cells is reversibly halted with a small-molecule drug. Approaches to increase the activity of chimeric antigen receptor (CAR)-T cells against solid tumors may also increase the risk of toxicity and other side effects. To improve the safety of CAR-T-cell therapy, we computationally designed a chemically disruptable heterodimer (CDH) based on the binding of two human proteins. The CDH self-assembles, can be disrupted by a small-molecule drug and has a high-affinity protein interface with minimal amino acid deviation from wild-type human proteins. We incorporated the CDH into a synthetic heterodimeric CAR, called STOP-CAR, that has an antigen-recognition chain and a CD3 zeta- and CD28-containing endodomain signaling chain. We tested STOP-CAR-T cells specific for two antigens in vitro and in vivo and found similar antitumor activity compared to second-generation (2G) CAR-T cells. Timed administration of the small-molecule drug dynamically inactivated the activity of STOP-CAR-T cells. Our work highlights the potential for structure-based design to add controllable elements to synthetic cellular therapies.Biotechnology & Applied Microbiologymembrane antigenbh3-only proteinsb-cellprostateinhibitorbindingsystemtext::journal::journal article::research article