Abstract
Kappa opioid receptor (KOR) agonists produce analgesic and anti-pruritic effects, but their clinical application was limited by dysphoria and hallucinations. Nalfurafine, a clinically used KOR agonist, does not cause dysphoria or hallucinations at therapeutic doses in humans. We found that in CD-1 mice nalfurafine produced analgesic and anti-scratch effects dose-dependently, like the prototypic KOR agonist U50,488H. In contrast, unlike U50,488H, nalfurafine caused no aversion, anhedonia, or sedation or and a low level of motor incoordination at the effective analgesia and anti-scratch doses. Thus, we established a mouse model that recapitulated important aspects of the clinical observations. We then employed a phosphoproteomics approach to investigate mechanisms underlying differential KOR-mediated effects. A large-scale mass spectrometry (MS)-based analysis on brains revealed that nalfurafine perturbed phosphoproteomes differently from U50,488H in a brain-region specific manner after 30-min treatment. In particular, U50,488H and nalfurafine imparted phosphorylation changes to proteins found in different cellular components or signaling pathways in different brain regions. Notably, we observed that U50,488H, but not nalfurafine, activated the mammalian target of rapamycin (mTOR) pathway in the striatum and cortex. Inhibition of the mTOR pathway by rapamycin abolished U50,488H-induced aversion, without affecting analgesic, anti-scratch, and sedative effects and motor incoordination. The results indicate that the mTOR pathway is involved in KOR agonist-induced aversion. This is the first demonstration that phosphoproteomics can be applied to agonist-specific signaling of G protein-coupled receptors (GPCRs) in mouse brains to unravel pharmacologically important pathways. Furthermore, this is one of the first two reports that the mTOR pathway mediates aversion caused by KOR activation.
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Acknowledgements
The expert assistance of Gaby Sowa, Igor Paron and Korbinian Mayr with the MS measurement is greatly appreciated. We thank Dr. Fred Ehlert of University of California, Irvine for calculating bias factors of KOR agonists. This work was supported by NIH grants R01 DA041359 and P30 DA013429 (L.-Y.L-C), and T32DA007237 (K.M.D.). The proteomic part of this work was funded by the Max-Planck Society for the Advancement of Science. J.J.L. was supported by an EMBO long-term fellowship.
Author contributions
Perform phosphoproteomics experiments and analyze data: J.J.L., M.M. design and perform animal treatment, behavioral experiments and analyze data: Y.T.C., K.M.D., C.C., P.H., T.A.G., J.W.M., A.C. and L.-Y.L.-C. Write manuscript: J.J.L., L.-Y.L.-C., Y.-T.C., P.H. provide inputs to manuscript: M.M., C.C., K.M.D.
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Liu, J.J., Chiu, YT., DiMattio, K.M. et al. Phosphoproteomic approach for agonist-specific signaling in mouse brains: mTOR pathway is involved in κ opioid aversion. Neuropsychopharmacol. 44, 939–949 (2019). https://doi.org/10.1038/s41386-018-0155-0
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DOI: https://doi.org/10.1038/s41386-018-0155-0
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