Strategies for expanding the sensor space of designer receptors are urgently needed to tailor cell-based therapies to respond to any type of medically relevant molecules. Here, we describe a universal approach to designing receptor scaffolds that enables antibody-specific molecular input to activate JAK/STAT, MAPK, PLCG or PI3K/Akt signaling rewired to transgene expression driven by synthetic promoters. To demonstrate its scope, we equipped the GEMS (generalized extracellular molecule sensor) platform with antibody fragments targeting a synthetic azo dye, nicotine, a peptide tag and the PSA (prostate-specific antigen) biomarker, thereby covering inputs ranging from small molecules to proteins. These four GEMS devices provided robust signaling and transgene expression with high signal-to-noise ratios in response to their specific ligands. The sensitivity of the nicotine- and PSA-specific GEMS devices matched the clinically relevant concentration ranges, and PSA-specific GEMS were able to detect pathological PSA levels in the serum of patients diagnosed with prostate cancer.
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Lim, W. A. & June, C. H. The principles of engineering immune cells to treat cancer. Cell 168, 724–740 (2017).
Brenner, M., Cho, J. H. & Wong, W. W. Synthetic biology: sensing with modular receptors. Nat. Chem. Biol. 13, 131–132 (2017).
Schukur, L., Geering, B., Charpin-El Hamri, G. & Fussenegger, M. Implantable synthetic cytokine converter cells with AND-gate logic treat experimental psoriasis. Sci. Transl. Med. 7, 318ra201 (2015).
Saxena, P., Charpin-El Hamri, G., Folcher, M., Zulewski, H. & Fussenegger, M. Synthetic gene network restoring endogenous pituitary-thyroid feedback control in experimental Graves’ disease. Proc. Natl. Acad. Sci. USA 113, 1244–1249 (2016).
Ye, H. et al. Pharmaceutically controlled designer circuit for the treatment of the metabolic syndrome. Proc. Natl. Acad. Sci. USA 110, 141–146 (2013).
Sadelain, M., Brentjens, R. & Rivière, I. The basic principles of chimeric antigen receptor design. Cancer Discov. 3, 388–398 (2013).
Morsut, L. et al. Engineering customized cell sensing and response behaviors using synthetic notch receptors. Cell 164, 780–791 (2016).
Kojima, R., Scheller, L. & Fussenegger, M. Nonimmune cells equipped with T-cell-receptor-like signaling for cancer cell ablation. Nat. Chem. Biol. 14, 42–49 (2017).
Schwarz, K. A., Daringer, N. M., Dolberg, T. B. & Leonard, J. N. Rewiring human cellular input-output using modular extracellular sensors. Nat. Chem. Biol. 13, 202–209 (2017).
Arber, C., Young, M. & Barth, P. Reprogramming cellular functions with engineered membrane proteins. Curr. Opin. Biotechnol. 47, 92–101 (2017).
Kawahara, M. & Nagamune, T. Engineering of mammalian cell membrane proteins. Curr. Opin. Chem. Eng. 1, 411–417 (2012).
Seubert, N. et al. Active and inactive orientations of the transmembrane and cytosolic domains of the erythropoietin receptor dimer. Mol. Cell 12, 1239–1250 (2003).
Pang, X. & Zhou, H. X. A common model for cytokine receptor activation: combined scissor-like rotation and self-rotation of receptor dimer induced by class I cytokine. PLoS Comput. Biol. 8, e1002427 (2012).
Banaszynski, L. A., Liu, C. W. & Wandless, T. J. Characterization of the FKBP.rapamycin.FRB ternary complex. J. Am. Chem. Soc. 127, 4715–4721 (2005).
Liu, W., Kawahara, M., Ueda, H. & Nagamune, T. Construction of a fluorescein-responsive chimeric receptor with strict ligand dependency. Biotechnol. Bioeng. 101, 975–984 (2008).
Spinelli, S., Tegoni, M., Frenken, L., van Vliet, C. & Cambillau, C. Lateral recognition of a dye hapten by a llama VHH domain. J. Mol. Biol. 311, 123–129 (2001).
Silver, J. S. & Hunter, C. A. gp130 at the nexus of inflammation, autoimmunity, and cancer. J. Leukoc. Biol. 88, 1145–1156 (2010).
Middleton, S. A. et al. Identification of a critical ligand binding determinant of the human erythropoietin receptor. Evidence for common ligand binding motifs in the cytokine receptor family. J. Biol. Chem. 271, 14045–14054 (1996).
Reichhart, E., Ingles-Prieto, A., Tichy, A. M., McKenzie, C. & Janovjak, H. A phytochrome sensory domain permits receptor activation by red light. Angew. Chem. Int. Ed. Engl. 55, 6339–6342 (2016).
Keeley, M. B., Busch, J., Singh, R. & Abel, T. TetR hybrid transcription factors report cell signaling and are inhibited by doxycycline. Biotechniques 39, 529–536 (2005).
Fussenegger, M. et al. Streptogramin-based gene regulation systems for mammalian cells. Nat. Biotechnol. 18, 1203–1208 (2000).
Abhinand, C. S., Raju, R., Soumya, S. J., Arya, P. S. & Sudhakaran, P. R. VEGF-A/VEGFR2 signaling network in endothelial cells relevant to angiogenesis. J. Cell Commun. Signal. 10, 347–354 (2016).
Tars, K. et al. Different binding modes of free and carrier-protein-coupled nicotine in a human monoclonal antibody. J. Mol. Biol. 415, 118–127 (2012).
Benowitz, N. L., Hukkanen, J. & Jacob, P. III. Nicotine chemistry, metabolism, kinetics and biomarkers. Handb. Exp. Pharmacol. 192, 29–60 (2009).
Tanenbaum, M. E., Gilbert, L. A., Qi, L. S., Weissman, J. S. & Vale, R. D. A protein-tagging system for signal amplification in gene expression and fluorescence imaging. Cell 159, 635–646 (2014).
Attard, G. et al. Prostate cancer. Lancet 387, 70–82 (2016).
Ménez, R. et al. Crystal structure of a ternary complex between human prostate-specific antigen, its substrate acyl intermediate and an activating antibody. J. Mol. Biol. 376, 1021–1033 (2008).
Stura, E. A. et al. Crystal structure of human prostate-specific antigen in a sandwich antibody complex. J. Mol. Biol. 414, 530–544 (2011).
Polascik, T. J., Oesterling, J. E. & Partin, A. W. Prostate specific antigen: a decade of discovery–what we have learned and where we are going. J. Urol. 162, 293–306 (1999).
Cookson, M. S. et al. Variation in the definition of biochemical recurrence in patients treated for localized prostate cancer: the American Urological Association Prostate Guidelines for Localized Prostate Cancer Update Panel report and recommendations for a standard in the reporting of surgical outcomes. J. Urol. 177, 540–545 (2007).
Ornitz, D. M. & Itoh, N. The fibroblast growth factor signaling pathway. Wiley Interdiscip. Rev. Dev. Biol. 4, 215–266 (2015).
Hall, M. P. et al. Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem. Biol. 7, 1848–1857 (2012).
Pogson, M., Parola, C., Kelton, W. J., Heuberger, P. & Reddy, S. T. Immunogenomic engineering of a plug-and-(dis)play hybridoma platform. Nat. Commun. 7, 12535 (2016).
Roybal, K. T. et al. Precision tumor recognition by T cells with combinatorial antigen-sensing circuits. Cell 164, 770–779 (2016).
Pollizzi, K. N. & Powell, J. D. Integrating canonical and metabolic signalling programmes in the regulation of T cell responses. Nat. Rev. Immunol. 14, 435–446 (2014).
Levy, D. E. & Darnell, J. E. Jr. Stats: transcriptional control and biological impact. Nat. Rev. Mol. Cell Biol. 3, 651–662 (2002).
Derynck, R. & Zhang, Y. E. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature 425, 577–584 (2003).
Falke, J. J. & Kim, S. H. Structure of a conserved receptor domain that regulates kinase activity: the cytoplasmic domain of bacterial taxis receptors. Curr. Opin. Struct. Biol. 10, 462–469 (2000).
Lombana, T. N. et al. Allosteric activation mechanism of the Mycobacterium tuberculosis receptor Ser/Thr protein kinase, PknB. Structure 18, 1667–1677 (2010).
Sasajima, Y., Aburatani, T., Sakamoto, K. & Ueda, H. Detection of protein tyrosine phosphorylation by open sandwich fluoroimmunoassay. Biotechnol. Prog. 22, 968–973 (2006).
We thank C. Rentsch and H. Püschel (University Hospital of Basel) for providing patient samples. We thank M. Xie and L. Schukur for generous advice and S. Reddy as well as A.C. Waindok (D-BSSE, ETH Zurich) for providing and transfecting WEN1.3 cells. This work was supported by the National Centre of Competence in Research (NCCR) Molecular Systems Engineering.
The authors declare no competing interests.
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Scheller, L., Strittmatter, T., Fuchs, D. et al. Generalized extracellular molecule sensor platform for programming cellular behavior. Nat Chem Biol 14, 723–729 (2018). https://doi.org/10.1038/s41589-018-0046-z
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