A novel G protein-biased agonist at the μ opioid receptor induces substantial receptor desensitisation through G protein-coupled receptor kinase.

Publisher: Wiley Country of Publication: England NLM ID: 7502536 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1476-5381 (Electronic) Linking ISSN: 00071188 NLM ISO Abbreviation: Br J Pharmacol Subsets: MEDLINE

Publication: London : Wiley
Original Publication: London, Macmillian Journals Ltd.

Analgesics, Opioid*/pharmacology ; Receptors, Opioid, mu*/metabolism; Rats ; Humans ; Animals ; GTP-Binding Proteins/metabolism ; Morphine/pharmacology ; Signal Transduction ; G-Protein-Coupled Receptor Kinases/metabolism ; Mammals/metabolism

Background and Purpose: G protein-biased μ opioid receptor agonists have the potential to induce less receptor desensitisation and tolerance than balanced opioids. Here, we investigated if the cyclic endomorphin analogue Tyr-c[D-Lys-Phe-Tyr-Gly] (Compound 1) is a G protein-biased μ agonist and characterised its ability to induce rapid receptor desensitisation in mammalian neurones.
Experimental Approach: The signalling and trafficking properties of opioids were characterised using bioluminescence resonance energy transfer assays, enzyme-linked immunosorbent assay and phosphosite-specific immunoblotting in human embryonic kidney 293 cells. Desensitisation of opioid-induced currents were studied in rat locus coeruleus neurones using whole-cell patch-clamp electrophysiology. The mechanism of Compound 1-induced μ receptor desensitisation was probed using kinase inhibitors.
Key Results: Compound 1 has similar intrinsic activity for G protein signalling as morphine. As predicted for a G protein-biased μ agonist, Compound 1 induced minimal agonist-induced internalisation and phosphorylation at intracellular μ receptor serine/threonine residues known to be involved in G protein-coupled receptor kinase (GRK)-mediated desensitisation. However, Compound 1 induced robust rapid μ receptor desensitisation in locus coeruleus neurons, to a greater degree than morphine. The extent of Compound 1-induced desensitisation was unaffected by activation or inhibition of protein kinase C (PKC) but was significantly reduced by inhibition of GRK.
Conclusion and Implications: Compound 1 is a novel G protein-biased μ agonist that induces substantial rapid receptor desensitisation in mammalian neurons. Surprisingly, Compound 1-induced desensitisation was demonstrated to be GRK dependent despite its G protein bias. Our findings refute the assumption that G protein-biased agonists will evade receptor desensitisation and tolerance.
Linked Articles: This article is part of a themed issue on Advances in Opioid Pharmacology at the Time of the Opioid Epidemic. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.7/issuetoc.
(© 2020 The British Pharmacological Society.)

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British Pharmacological Society (BPS); R01 DA007315 United States DA NIDA NIH HHS; R01 DA036975 United States DA NIDA NIH HHS

Keywords: G protein-coupled receptor kinases; arrestins; biased agonism; electrophysiology; opiates; opioids; receptor desensitisation

0 (Analgesics, Opioid)
0 (Receptors, Opioid, mu)
EC 3.6.1.- (GTP-Binding Proteins)
76I7G6D29C (Morphine)
EC 2.7.11.16 (G-Protein-Coupled Receptor Kinases)

Date Created: 20201127 Date Completed: 20230303 Latest Revision: 20230314

20240829

10.1111/bph.15334

33245558