Uncovering Non-Monotonic Antagonistic and Synergistic Combinations (UNMASC), a robust method with applications to T cell differentiation in vitro

Induced pluripotent stem cells (iPSCs) have been proposed as an alternative T cell source for CAR-T cell therapies, as they can be differentiated and matured into T cells in vitro using cytokines. These assays benefit from computational and mathematical models to design appropriate experimental protocols. However, models are limited by typical monotonic dose-responses, preventing them from being used for cytokine effects that are largely non-monotonic. To address this shortcoming, we developed Uncovering Non-Monotonic Antagonistic and Synergistic Combinations (UNMASC), a novel mathematical model describing non-monotonic dose-response surfaces of cytokine interactions that distinguishes synergy of efficacy and potency. We showed that our approach successfully recapitulates non-monotonic observed dose-response surfaces characterizing in vitro T cell progenitor differentiation. Our results highlighted cytokine combinations with antagonistic effects alone but synergistic in combination, particularly IL3 and IL7. Together, UNMASC accelerates the efficient cell generation assays and extends drug interaction surfaces to a broader range of dose-responses.