Cell fate-inducing CARs orthogonally control multiple signaling pathways.

While chimeric antigen receptor (CAR) has been clinically applied in T cell-mediated cancer therapy, CARs that combinatorially control general cell fates have yet to be practical. In this study, we aim to control multiple cell fates simultaneously by combinatorial activation of multiple cell fate-inducing CARs (cfiCARs). We construct CARs that transduce two different signals using two antigen-specific CARs. The CARs are co-expressed pairwise on the cell surface to let the two antigens act alone or in combination, thus arbitrarily control multiple cell fates. We utilize signaling domains derived from natural receptors (gp130, receptor activator of nuclear factor κB [RANK], c-Fms, and Fas) and tyrosine motif-engineered artificial signaling domains that preferentially recruit target signaling molecules (Shc and STAT3) for inducing specific cell fates by the CARs. Consequently, the signaling molecules downstream of these receptors are activated orthogonally by the corresponding CAR-specific antigens. Furthermore, two completely different cell fates (proliferation and death) are successfully controlled simultaneously or sequentially over time by adding the two antigens, demonstrating proliferation- and apoptosis-inducing CARs (piCAR and aiCAR). Thus, cfiCARs will be applied for delineating the basic principle of cell fate control and improving the efficacy of cell therapy, which would significantly contribute to cell biology and future medicine.