Conversion of natural cytokine receptors into orthogonal synthetic biosensors

Synthetic receptors enable bioengineers to build cell-based therapies that perform therapeutic functions in a targeted or conditional fashion to enhance specificity and efficacy. Although many synthetic receptors exist, it remains challenging to generate new receptors that sense soluble cues and relay that detection through orthogonal mechanisms independent of native pathways. Here we co-opt natural cytokine receptor ectodomains into modular extracellular sensor architecture (MESA) receptors to form natural ectodomain (NatE) MESA receptors. We generated multiple functional, orthogonal synthetic cytokine receptors, identified design principles and constraints and propose guidance for extending this approach to other natural receptors. We demonstrate the utility of NatE MESA by engineering T cells to sense an immunosuppressive cue and respond with customized transcriptional output to support chimeric antigen receptor T cell activity. Lastly, we multiplex NatE MESA to logically evaluate multiple cues associated with the tumor microenvironment. These technologies and learnings will enable engineering cellular functions for new applications.