Inhibition of sensory neuron driven acute, inflammatory, and neuropathic pain using a humanised chemogenetic system

Sanchez, J. P.; Middleton, S. J.; Pattison, L. A.; Hilton, H.; Awadelkareem, M. A.; Zuberi, S. R.; Renke, M. B.; Hu, H.; Yang, X.; Clark, A. J.; Smith, E. S. J.; Bennett, D. L.

doi:10.1101/2023.03.21.533690

bioRxiv · 10.1101/2023.03.21.533690

Inhibition of sensory neuron driven acute, inflammatory, and neuropathic pain using a humanised chemogenetic system

Sanchez, J. P.·Middleton, S. J.·Pattison, L. A.·Hilton, H.·Awadelkareem, M. A.·Zuberi, S. R.·Renke, M. B.·Hu, H.·Yang, X.·Clark, A. J.·Smith, E. S. J.·Bennett, D. L.

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Abstract

Hyperexcitability in sensory neurons is known to underlie many of the maladaptive changes associated with persistent pain. Chemogenetics has shown promise as a means to suppress such excitability, yet chemogenetic approaches suitable for human applications are needed. PSAM4-GlyR is a modular system based on the human 7 nicotinic acetylcholine and glycine receptors, which responds to inert chemical ligands and the clinically-approved drug, varenicline. Here, we demonstrated the efficacy of this channel in silencing both mouse and human sensory neurons by the activation of large shunting conductances after agonist administration. Virally-mediated expression of PSAM4-GlyR in mouse sensory neurons produced behavioural hyposensitivity upon agonist administration, which was recovered upon agonist washout. Importantly, stable expression of the channel led to similar reversible behavioural effects even after 10 months of viral delivery. Mechanical and spontaneous pain readouts were also ameliorated by PSAM4-GlyR activation in acute and joint pain inflammation models. Furthermore, suppression of mechanical hypersensitivity generated by a spared nerve injury model of neuropathic pain was also observed upon activation of the channel. Effective silencing of behavioural hypersensitivity was reproduced in a human model of hyperexcitability and clinical pain: PSAM4-GlyR activation decreased the excitability of human induced pluripotent stem-cell-derived sensory neurons and spontaneous activity due to a gain of function NaV1.7 mutation causing inherited erythromelalgia. Our results demonstrate the contribution of sensory neuron hyperexcitability to neuropathic pain and the translational potential of an effective, stable and reversible human-based chemogenetic system for the treatment of pain.

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Sanchez, J. P., Middleton, S. J., Pattison, L. A., Hilton, H., Awadelkareem, M. A., Zuberi, S. R., Renke, M. B., Hu, H., Yang, X., Clark, A. J., Smith, E. S. J., Bennett, D. L.. 2023-03-21. Inhibition of sensory neuron driven acute, inflammatory, and neuropathic pain using a humanised chemogenetic system. https://doi.org/10.1101/2023.03.21.533690

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