Current treatment options for chronic pain are often associated with dose-limiting toxicities, or lead to drug tolerance or addiction. Here, we describe a pain management strategy, based on cell-engineering principles and inspired by synthetic biology, consisting of microencapsulated human designer cells that produce huwentoxin-IV (a safe and potent analgesic peptide that selectively inhibits the pain-triggering voltage-gated sodium channel NaV1.7) in response to volatile spearmint aroma and in a dose-dependent manner. Spearmint sensitivity was achieved by ectopic expression of the R-carvone-responsive olfactory receptor OR1A1 rewired via an artificial G-protein deflector to induce the expression of a secretion-engineered and stabilized huwentoxin-IV variant. In a model of chronic inflammatory and neuropathic pain, mice bearing the designer cells showed reduced pain-associated behaviour on oral intake or inhalation-based intake of spearmint essential oil, and absence of cardiovascular, immunogenic and behavioural side effects. Our proof-of-principle findings indicate that therapies based on engineered cells can achieve robust, tunable and on-demand analgesia for the long-term management of chronic pain.
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Lampert, A., O’Reilly, A. O., Reeh, P. & Leffler, A. Sodium channelopathies and pain. Pflug. Arch. Eur. J. Phy. 460, 249–263 (2010).
King, G. F. & Vetter, I. No gain, no pain: NaV1.7 as an analgesic target. ACS Chem. Neurosci. 5, 749–751 (2014).
Goins, W. F., Cohen, J. B. & Glorioso, J. C. Gene therapy for the treatment of chronic peripheral nervous system pain. Neurobiol. Dis. 48, 255–270 (2012).
Dib-Hajj, S. D., Yang, Y., Black, J. A. & Waxman, S. G. The NaV1.7 sodium channel: from molecule to man. Nat. Rev. Neurosci. 14, 49–62 (2013).
Grosser, T., Woolf, C. J. & FitzGerald, G. A. Time for nonaddictive relief of pain. Science 355, 1026–1027 (2017).
Adams, D. J., Callaghan, B. & Berecki, G. Analgesic conotoxins: block and G protein-coupled receptor modulation of N-type (CaV2.2) calcium channels. Br. J. Pharmacol. 166, 486–500 (2012).
McGivern, J. G. Ziconotide: a review of its pharmacology and use in the treatment of pain. Neuropsychiatr. Dis. Treat. 3, 69–85 (2007).
Revell, J. D. et al. Potency optimization of Huwentoxin-IV on hNav1.7: a neurotoxin TTX-S sodium-channel antagonist from the venom of the Chinese bird-eating spider Selenocosmia huwena. Peptides 44, 40–46 (2013).
Murray, J. K. et al. Engineering potent and selective analogues of GpTx-1, a tarantula venom peptide antagonist of the NaV1.7 sodium channel. J. Med. Chem. 58, 2299–2314 (2015).
Liu, Y. et al. Analgesic effects of Huwentoxin-IV on animal models of inflammatory and neuropathic pain. Protein Pept. Lett. 21, 153–158 (2014).
Ye, H. et al. Self-adjusting synthetic gene circuit for correcting insulin resistance. Nat. Biomed. Eng. 1, 1–9 (2016).
Chowdhury, S. et al. Discovery of XEN907, a spirooxindole blocker of NaV1.7 for the treatment of pain. Bioorg. Med. Chem. Lett. 21, 3676–3681 (2011).
McCormack, K. et al. Voltage sensor interaction site for selective small molecule inhibitors of voltage-gated sodium channels. Proc. Natl Acad. Sci. USA 110, E2724–E2732 (2013).
Sheets, P. L., Jarecki, B. W. & Cummins, T. R. Lidocaine reduces the transition to slow inactivation in Nav1.7 voltage-gated sodium channels. Br. J. Pharmacol. 164, 719–730 (2011).
Gintant, G., Sager, P. T. & Stockbridge, N. Evolution of strategies to improve preclinical cardiac safety testing. Nat. Rev. Drug Discov. 15, 457–471 (2016).
Lee, J. H. et al. A monoclonal antibody that targets a NaV1.7 channel voltage sensor for pain and itch relief. Cell 157, 1393–1404 (2014).
Vissers, K. C., Geenen, F., Biermans, R. & Meert, T. F. Pharmacological correlation between the formalin test and the neuropathic pain behavior in different species with chronic constriction injury. Pharmacol. Biochem Behav. 84, 479–486 (2006).
Hunskaar, S., Fasmer, O. B. & Hole, K. Formalin test in mice, a useful technique for evaluating mild analgesics. J. Neurosci. Methods 14, 69–76 (1985).
Sufka, K. J., Watson, G. S., Nothdurft, R. E. & Mogil, J. S. Scoring the mouse formalin test: validation study. Eur. J. Pain. 2, 351–358 (1998).
Dahoun, T., Grasso, L., Vogel, H. & Pick, H. Recombinant expression and functional characterization of mouse olfactory receptor mOR256-17 in mammalian cells. Biochemistry 50, 7228–7235 (2011).
Sun, N., Lee, A. & Wu, J. C. Long term non-invasive imaging of embryonic stem cells using reporter genes. Nat. Protoc. 4, 1192–1201 (2009).
Saito, H., Kubota, M., Roberts, R. W., Chi, Q. & Matsunami, H. RTP family members induce functional expression of mammalian odorant receptors. Cell 119, 679–691 (2004).
Reichling, J., Schnitzler, P., Suschke, U. & Saller, R. Essential oils of aromatic plants with antibacterial, antifungal, antiviral, and cytotoxic properties—an overview. Forsch. Komplementmed. 16, 79–90 (2009).
Muller, M. et al. Designed cell consortia as fragrance-programmable analog-to-digital converters. Nat. Chem. Biol. 13, 309–316 (2017).
Ni, C. H. et al. The anxiolytic effect of aromatherapy on patients awaiting ambulatory surgery: a randomized controlled trial. Evid. Based Compl. Alt. 2013, 927419 (2013).
Bilia, A. R. et al. Essential oils loaded in nanosystems: a developing strategy for a successful therapeutic approach. Evid. Based Compl. Alt. 2014, 651593 (2014).
Xu, Z. Z. et al. Resolvins RvE1 and RvD1 attenuate inflammatory pain via central and peripheral actions. Nat. Med. 16, 592–597 (2010).
Liu, C., Cao, J., Ren, X. & Zang, W. Nav1.7 protein and mRNA expression in the dorsal root ganglia of rats with chronic neuropathic pain. Neural Regen. Res. 7, 1540–1544 (2012).
Burgess, G. & Williams, D. The discovery and development of analgesics: new mechanisms, new modalities. J. Clin. Investig. 120, 3753–3759 (2010).
Cox, J. J. et al. An SCN9A channelopathy causes congenital inability to experience pain. Nature 444, 894–898 (2006).
Xie, M. & Fussenegger, M. Mammalian designer cells: engineering principles and biomedical applications. Biotechnol. J. 10, 1005–1018 (2015).
Trounson, A. & DeWitt, N. D. Pluripotent stem cells progressing to the clinic. Nat. Rev. Mol. Cell Biol. 17, 194–200 (2016).
Lathuiliere, A., Cosson, S., Lutolf, M. P., Schneider, B. L. & Aebischer, P. A high-capacity cell macroencapsulation system supporting the long-term survival of genetically engineered allogeneic cells. Biomaterials 35, 779–791 (2014).
Prohaska, J. R. & Broderius, M. Plasma peptidylglycine alpha-amidating monooxygenase (PAM) and ceruloplasmin are affected by age and copper status in rats and mice. Comp. Biochem. Phys. B 143, 360–366 (2006).
Merkler, D. J. C-terminal amidated peptides: production by the in vitro enzymatic amidation of glycine-extended peptides and the importance of the amide to bioactivity. Enzym. Microb. Technol. 16, 450–456 (1994).
Xie, M. et al. beta-cell-mimetic designer cells provide closed-loop glycemic control. Science 354, 1296–1301 (2016).
Bonin, R. P., Bories, C. & De Koninck, Y. A simplified up-down method (SUDO) for measuring mechanical nociception in rodents using von Frey filaments. Mol. Pain. 10, 26 (2014).
Denk, F., Crow, M., Didangelos, A., Lopes, D. M. & McMahon, S. B. Persistent alterations in microglial enhancers in a model of chronic pain. Cell Rep. 15, 1771–1781 (2016).
Hassan, A. M. et al. Visceral hyperalgesia caused by peptide YY deletion and Y2 receptor antagonism. Sci. Rep. 7, 40968 (2017).
Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M. & Yaksh, T. L. Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods 53, 55–63 (1994).
Ren, K. & Dubner, R. Inflammatory models of pain and hyperalgesia. ILAR J. 40, 111–118 (1999).
Beko, K. et al. Contribution of platelet P2Y12 receptors to chronic Complete Freund’s adjuvant-induced inflammatory pain. J. Thromb. Haemost. 15, 1223–1235 (2017).
Bennett, G. J. & Xie, Y. K. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 33, 87–107 (1988).
Motulsky, H. J. & Brown, R. E. Detecting outliers when fitting data with nonlinear regression—a new method based on robust nonlinear regression and the false discovery rate. BMC Bioinformatics 7, 123 (2006).
Wang, H., Ye, H., Xie, M., Daoud El-Baba, M. & Fussenegger, M. Cosmetics-triggered percutaneous remote control of transgene expression in mice. Nucleic Acids Res. 43, e91 (2015).
Branson, K. Distinguishing seemingly indistinguishable animals with computer vision. Nat. Methods 11, 721–722 (2014).
We thank L. Scheller for critical comments on the manuscript; A. W. Xie (Welfine Science & Technology) for providing the Bel-Air aroma diffuser; V. Jäggin and T. Lopes for assistance with fluorescence-activated cell sorting; M. Daoud-El Baba, S. Xue and J. Yin for help with animal experiments, and Y. Huang (ChemPartner) for the CCI-based mouse model; D. Bodenham for support with statistical analysis; and M. Müller, P. Saxena and R. Kojima for generous advice. This work was supported by a European Research Council advanced grant (ProNet, no. 321381), by the National Centre of Competence in Research Molecular Systems Engineering, the National Natural Science Foundation of China (grant nos 31522017, 31470834 and 31670869) and the Thousand Youth Talents Plan of China.
The authors declare no competing financial interests.
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Representative 13-second video of Straub tail and rearing reactions of mice treated with morphine.
Representative epileptic seizure of a CCI-mouse treated with Tramadol at day 7 (with 3 daily injections of 50 mg per kg).
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Wang, H., Xie, M., Charpin-El Hamri, G. et al. Treatment of chronic pain by designer cells controlled by spearmint aromatherapy. Nat Biomed Eng 2, 114–123 (2018). https://doi.org/10.1038/s41551-018-0192-3
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