Intraarticular Adeno-Associated Virus Serotype AAV-PHP.S-Mediated Chemogenetic Targeting of Knee-Innervating Dorsal Root Ganglion Neurons Alleviates Inflammatory Pain in Mice.
Rickman, Rebecca H
Heppenstall, Paul A
Smith, Ewan St John
Arthritis & rheumatology (Hoboken, N.J.)
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Chakrabarti, S., Pattison, L. A., Doleschall, B., Rickman, R. H., Blake, H., Callejo, G., Heppenstall, P. A., & et al. (2020). Intraarticular Adeno-Associated Virus Serotype AAV-PHP.S-Mediated Chemogenetic Targeting of Knee-Innervating Dorsal Root Ganglion Neurons Alleviates Inflammatory Pain in Mice.. Arthritis & rheumatology (Hoboken, N.J.), 72 (10), 1749-1758. https://doi.org/10.1002/art.41314
Objective: Joint pain is the major clinical symptom of arthritis that affects millions of people. Controlling the excitability of knee-innervating dorsal root ganglion (DRG) neurons (knee neurons) could potentially provide pain relief. Therefore, our objective was to evaluate whether the newly engineered adeno-associated virus (AAV) serotype, AAV-PHP.S, can deliver functional artificial receptors to control knee neuron excitability following intra-articular knee injection. Methods: AAV-PHP.S virus packaged with dTomato fluorescent protein and either excitatory (Gq) or inhibitory (Gi) designer receptors activated by designer drugs (DREADDs) was injected into the knee joint of adult mice. Labelling of DRG neurons by AAV-PHP.S from the knee was evaluated using immunohistochemistry. Functionality of Gq- and Gi-DREADDs was evaluated using whole-cell patch clamp electrophysiology on acutely cultured DRG neurons. Pain behavior in mice was assessed using a digging assay, dynamic weight bearing and rotarod, before and after intra-peritoneal administration of the DREADD activator, Compound 21. Results: We show that AAV-PHP.S can deliver functional genes into the DRG neurons when injected into the knee joint in a similar manner to the well-established retrograde tracer, fast blue. Short-term activation of AAV-PHP.S delivered Gq-DREADD increases excitability of knee neurons in vitro, without inducing overt pain in mice when activated in vivo. By contrast, in vivo Gi-DREADD activation alleviated complete Freund’s adjuvant mediated knee inflammation-induced deficits in digging behavior, with a concomitant decrease in knee neuron excitability observed in vitro. Conclusions We describe an AAV-mediated chemogenetic approach to specifically control joint ¬pain, which may be utilized in translational arthritic pain research.
Knee Joint, Ganglia, Spinal, Neurons, Animals, Mice, Dependovirus, Pain, Disease Models, Animal, Inflammation, Pain Management, Genetic Therapy
Is supplemented by: https://doi.org/10.17863/CAM.46171
This study was supported by Versus Arthritis Project grants (RG 20930 and RG 21973) to E. St. J. S. and G.C. S.C. was supported by Gates Cambridge Trust scholarship, Cambridge Philosophical Society research fund, Department of Pharmacology (University of Cambridge) travel award and Corpus Christi College (University of Cambridge) travel award. L.A.P. was supported by the University of Cambridge BBSRC Doctoral Training Programme (BB/M011194/1). P.A.H and B.D. were supported by EMBL.
ARTHRITIS RESEARCH UK (20930)
Arthritis Research UK (11600/21973)
External DOI: https://doi.org/10.1002/art.41314
This record's URL: https://www.repository.cam.ac.uk/handle/1810/305329
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