Mediation of opioid analgesia by a truncated 6-transmembrane GPCR
Zhigang Lu, … , Gavril W. Pasternak, Ying-Xian Pan
Zhigang Lu, … , Gavril W. Pasternak, Ying-Xian Pan
Published July 1, 2015; First published May 26, 2015
Citation Information: J Clin Invest. 2015;125(7):2626-2630. https://doi.org/10.1172/JCI81070.
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Brief Report Genetics Neuroscience Therapeutics

Mediation of opioid analgesia by a truncated 6-transmembrane GPCR

Abstract

The generation of potent opioid analgesics that lack the side effects of traditional opioids may be possible by targeting truncated splice variants of the μ-opioid receptor. μ-Opioids act through GPCRs that are generated from the Oprm1 gene, which undergoes extensive alternative splicing. The most abundant set of Oprm1 variants encode classical full-length 7 transmembrane domain (7TM) μ-opioid receptors that mediate the actions of the traditional μ-opioid drugs morphine and methadone. In contrast, 3-iodobenzoyl-6β-naltrexamide (IBNtxA) is a potent analgesic against thermal, inflammatory, and neuropathic pain that acts independently of 7TM μ-opioid receptors but has no activity in mice lacking a set of 6TM truncated μ-opioid receptor splice variants. Unlike traditional opioids, IBNtxA does not depress respiration or result in physical dependence or reward behavior, suggesting it acts through an alternative μ-opioid receptor target. Here we demonstrated that a truncated 6TM splice variant, mMOR-1G, can rescue IBNtxA analgesia in a μ-opioid receptor–deficient mouse that lacks all Oprm1 splice variants, ablating μ-opioid activity in these animals. Intrathecal administration of lentivirus containing the 6TM variant mMOR-1G restored IBNtxA, but not morphine, analgesia in Oprm1-deficient animals. Together, these results confirm that a truncated 6TM GPCR is both necessary and sufficient for IBNtxA analgesia.

Authors

Zhigang Lu, Jin Xu, Grace C. Rossi, Susruta Majumdar, Gavril W. Pasternak, Ying-Xian Pan

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Figure 3

Lentivirus rescue of IBNtxA analgesia.

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Lentivirus rescue of IBNtxA analgesia. (A) Opioid analgesia. Analgesia w...
(A) Opioid analgesia. Analgesia was determined in groups of mice (n = 6–13) at the stated time. (*P < 0.0001 compared with week 0; ANOVA followed by Tukey). (B) IBNtxA cumulative dose–response curves were carried out in exon 1/exon 11 KO mice with lentivirus vector alone (n = 4), lenti–mMOR-1G (ED50 1.1 mg/kg [95% CI, 0.72–1.53], n = 18) and WT mice (ED50 0.42 mg/kg [95% CI, 0.29–0.58], n = 16). (C) Single doses of IBNtxA (2.5 mg/kg, s.c. n = 5), morphine (10 mg/kg, s.c. n = 7), fentanyl (0.08 mg/kg, s.c. n = 7), buprenorphine (1 mg/kg, s.c. n = 7), ketocyclazocine (2 mg/kg, s.c. n = 7), or levorphanol (0.8 mg/kg, s.c. n = 7) were administered to groups of either WT or exon 1/exon 11 KO mice infected with lenti–mMOR-1G. The mice were assessed for analgesia. Another group of mice received IBNtxA with levallorphan (2.5 mg/kg, s.c. n = 7). ANOVA shows that the IBNtxA group and the ketocyclazocine group were significantly different (*P < 0.001 and **P < 0.006 by ANOVA, respectively). The IBNtxA WT and lenti–mMOR-1G animals were not significantly different from each other but were different from both the exon 1/exon 11 KO alone and the lenti–mMOR‑1G/levallorphan groups (Tukey). Ketocyclazocine in WT and lenti–mMOR-1G were not significantly different from each other but were different from exon 1/exon 11 alone (Tukey).