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Rewiring human cellular input–output using modular extracellular sensors

Abstract

Engineered cell-based therapies comprise a promising emerging strategy for treating diverse diseases. Realizing this promise requires new tools for engineering cells to sense and respond to soluble extracellular factors, which provide information about both physiological state and the local environment. Here, we report such a biosensor engineering strategy, leveraging a self-contained receptor–signal transduction system termed modular extracellular sensor architecture (MESA). We developed MESA receptors that enable cells to sense vascular endothelial growth factor (VEGF) and, in response, secrete interleukin 2 (IL-2). By implementing these receptors in human T cells, we created a customized function not observed in nature—an immune cell that responds to a normally immunosuppressive cue (VEGF) by producing an immunostimulatory factor (IL-2). Because this platform utilizes modular, engineerable domains for ligand binding (antibodies) and output (programmable transcription factors based upon Cas9), this approach may be readily extended to novel inputs and outputs. This generalizable approach for rewiring cellular functions could enable both translational applications and fundamental biological research.

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Figure 1: VEGF-MESA receptor development.
Figure 2: VEGF-MESA receptor implementation.
Figure 3: Rewiring of cellular input–output using MESA.

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Acknowledgements

K.A.S. and N.M.D. were supported in part by the National Institutes of Health T32 Training Grant GM 008449 through Northwestern University's Biotechnology Training Program. T.B.D. was supported by the Department of Defense (DoD) through the National Defense Science & Engineering Graduate Fellowship (NDSEG) Program. This work was supported by the Northwestern University Flow Cytometry Facility and a Cancer Center Support Grant (NCI CA060553). This project was supported by the Defense Advanced Research Projects Agency, Award number W911NF-11-2-0066 (to J.N.L.). No funding sources had any involvement in the writing of this manuscript.

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K.A.S. performed, designed, and analyzed experiments. T.B.D. performed and analyzed experiments. K.A.S., N.M.D., and J.N.L. conceived of the study. K.A.S., T.B.D., and J.N.L. wrote the manuscript and prepared the figures.

Corresponding author

Correspondence to Joshua N Leonard.

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Competing interests

N.M.D., K.A.S. and J.N.L. are inventors on US patent applications covering inventions described in this manuscript.

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Supplementary Results, Supplementary Figures 1–10, Supplementary Table 1 and Supplementary Note. (PDF 1738 kb)

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Schwarz, K., Daringer, N., Dolberg, T. et al. Rewiring human cellular input–output using modular extracellular sensors. Nat Chem Biol 13, 202–209 (2017). https://doi.org/10.1038/nchembio.2253

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