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 kappa B [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.

Automated summary

This study aims to control multiple cell fates simultaneously by combinatorially activating multiple cell fate-inducing CARs (cfiCARs). Through the co-expression of two antigen-specific CARs, the CARs transduce two different signals, which can arbitrarily control multiple cell fates. By utilizing natural receptors and engineered signaling domains, specific cell fates can be induced by activating downstream signaling molecules orthogonally. The study successfully demonstrates the simultaneous or sequential control of proliferation and apoptosis, showing the potential of cfiCARs in cell fate control and cell therapy efficacy improvement.