Pharmacological Characterization of Receptor Redistribution and β-Arrestin Recruitment Assays for the Cannabinoid Receptor 1

Lee, Miranda M.C. van der; Blomenröhr, Marion; Doelen, Antoon A. van der; Wat, Jesse W.Y.; Smits, Niels; Hanson, Bonnie J.; Koppen, Chris J. van; Zaman, Guido J.R.

doi:10.1177/1087057109337937

Cited by 39 publications

(46 citation statements)

References 42 publications

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“…Recently published studies characterizing GPR55 activation using a related technology (␤-arrestin-PathHunter) or demonstrating that monitoring agonist-mediated receptor internalization and ␤-arrestin redistribution can recognize agonists, antagonists, and allosteric modulators support our findings (38,39). The preponderance of data emerging from particularly this and other reports (9 -11, 38) indicate that GPR55 is not activated by cannabinoid receptor ligands in a manner compatible with expected cannabinoid receptor pharmacology.…”

Section: Pharmacological Characterization Of Gpr55supporting

confidence: 71%

Atypical Responsiveness of the Orphan Receptor GPR55 to Cannabinoid Ligands

Kapur

Zhao

Sharir

et al. 2009

Journal of Biological Chemistry

251

289

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The cannabinoid receptor 1 (CB 1 ) and CB 2 cannabinoid receptors, associated with drugs of abuse, may provide a means to treat pain, mood, and addiction disorders affecting widespread segments of society. Whether the orphan G-protein coupled receptor GPR55 is also a cannabinoid receptor remains unclear as a result of conflicting pharmacological studies. GPR55 has been reported to be activated by exogenous and endogenous cannabinoid compounds but surprisingly also by the endogenous non-cannabinoid mediator lysophosphatidylinositol (LPI). We examined the effects of a representative panel of cannabinoid ligands and LPI on GPR55 using a ␤-arrestingreen fluorescent protein biosensor as a direct readout of agonist-mediated receptor activation. Our data demonstrate that AM251 and SR141716A (rimonabant), which are cannabinoid antagonists, and the lipid LPI, which is not a cannabinoid receptor ligand, are GPR55 agonists. They possess comparable efficacy in inducing ␤-arrestin trafficking and, moreover, activate the G-protein-dependent signaling of protein kinase C␤II. Conversely, the potent synthetic cannabinoid agonist CP55,940 acts as a GPR55 antagonist/partial agonist. CP55,940 blocks GPR55 internalization, the formation of ␤-arrestin GPR55 complexes, and the phosphorylation of ERK1/2; CP55,940 produces only a slight amount of protein kinase C␤II membrane recruitment but does not stimulate membrane remodeling like LPI, AM251, or rimonabant. Our studies provide a paradigm for measuring the responsiveness of GPR55 to a variety of ligand scaffolds comprising cannabinoid and novel compounds and suggest that at best GPR55 is an atypical cannabinoid responder. The activation of GPR55 by rimonabant may be responsible for some of the off-target effects that led to its removal as a potential obesity therapy.The CB 1 2 and CB 2 cannabinoid receptors comprise a twomember subfamily of G-protein-coupled receptors (GPCRs) that are notable as the targets of the tetrahydrocannabinol (THC) derivatives found in marijuana. More recently CB 1 receptors along with other GPCRs have been promoted as therapeutic pharmacological targets in the billion dollar weight loss market for controversial drugs such as rimonabant (SR141716A) and Fen-phen. Thus, an important utility of cannabinoid family receptors to society appears to arise from their role in regulating a broad spectrum of addiction-based behaviors, and the addition of new members to the cannabinoid receptor family may have social and economic implications that reach far beyond the initial scientific discovery. As a consequence, the re-classification of an orphan GPCR as a cannabinoid family member should be done with caution requiring strict criteria of receptor activation by THC derivatives or endogenous cannabinoid compounds and a widespread agreement of the results by the scientific community.Marijuana, one of the most widely abused substances (1), mediates many of its psychotropic effects by targeting CB 1 receptors in the central nervous system, but studies with CB 1 and CB 2 kno...

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Section: Pharmacological Characterization Of Gpr55supporting

confidence: 71%

Atypical Responsiveness of the Orphan Receptor GPR55 to Cannabinoid Ligands

Kapur

Zhao

Sharir

et al. 2009

Journal of Biological Chemistry

251

289

View full text Add to dashboard Cite

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“…Heteromerization is known to affect b-arrestin recruitment. b-arrestin2 (Jin et al, 1999;van der Lee et al, 2009) and b-arrestin1 facilitate the internalization of CB 1 after activation (Laprairie et al, 2014). We demonstrated that D 2 antagonism inhibited CB 1 agonist-induced recruitment of b-arrestin1 to CB 1 -D 2L -Ga s complexes and inhibited CB 1 receptor internalization in STHdh Q7/Q7 cells in a dosedependent manner.…”

Section: Discussionmentioning

confidence: 70%

“…CB 1 activation is followed by C-terminal tail phosphorylation and b-arrestin1 (Laprairie et al, 2014) or b-arrestin2 (Jin et al, 1999;van der Lee et al, 2009) recruitment to CB 1 , leading to receptor internalization. b-arrestin1 recruitment to CB 1 following ligand application was measured over 30 minutes using BRET 2 .…”

Section: Bretmentioning

confidence: 99%

Antagonism of Dopamine Receptor 2 Long Affects Cannabinoid Receptor 1 Signaling in a Cell Culture Model of Striatal Medium Spiny Projection Neurons

et al. 2016

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“…We also find that Org 27569 alone acts as a weak inverse agonist in the [ 35 S]GTPgS assay but apparently as an agonist of Ga s signaling and a weak partial agonist of Gai signaling in pERK assays, respectively. Furthermore we find that this compound alone has no effect on b-arrestin recruitment, which is known to be PTX-insensitive (van der Lee et al, 2009), but does act as an inhibitor of CB 1 agonist-induced b-arrestin recruitment. In line with findings of Ahn et al (2012), we also identify S.E.M.…”

Section: Effect Of Allosteric Modulators On Cb 1 Receptor Agonist Binmentioning

confidence: 74%

CB₁ Receptor Allosteric Modulators Display Both Agonist and Signaling Pathway Specificity

et al. 2012

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We have previously identified allosteric modulators of the cannabinoid CB 1 receptor (Org 27569, PSNCBAM-1) that display a contradictory pharmacological profile: increasing the specific binding of the CB 1 receptor agonist [ 3 H]CP55940 but producing a decrease in CB 1 receptor agonist efficacy. Here we investigated the effect one or both compounds in a broad range of signaling endpoints linked to CB 1 receptor activation. We assessed the effect of these compounds on CB 1 receptor agonist-induced [35 S]GTPgS binding, inhibition, and stimulation of forskolin-stimulated cAMP production, phosphorylation of extracellular signal-regulated kinases (ERK), and b-arrestin recruitment. We also investigated the effect of these allosteric modulators on CB 1 agonist binding kinetics. -mediated), and inhibition (Ga i -mediated) of cAMP production and b-arrestin recruitment. In contrast, it acts as an enhancer of agonist-induced ERK phosphorylation. Alone, the compound can act also as an allosteric agonist, increasing cAMP production and ERK phosphorylation. We find that in both saturation and kineticbinding experiments, the Org 27569 and PSNCBAM-1 appeared to influence only orthosteric ligand maximum occupancy rather than affinity. The data indicate that the allosteric modulators share a common mechanism whereby they increase available high-affinity CB 1 agonist binding sites. The receptor conformation stabilized by the allosterics appears to induce signaling and also selectively traffics orthosteric agonist signaling via the ERK phosphorylation pathway.

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Pharmacological Characterization of Receptor Redistribution and β-Arrestin Recruitment Assays for the Cannabinoid Receptor 1

Cited by 39 publications

References 42 publications

Atypical Responsiveness of the Orphan Receptor GPR55 to Cannabinoid Ligands

Atypical Responsiveness of the Orphan Receptor GPR55 to Cannabinoid Ligands

Antagonism of Dopamine Receptor 2 Long Affects Cannabinoid Receptor 1 Signaling in a Cell Culture Model of Striatal Medium Spiny Projection Neurons

CB₁ Receptor Allosteric Modulators Display Both Agonist and Signaling Pathway Specificity

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Pharmacological Characterization of Receptor Redistribution and β-Arrestin Recruitment Assays for the Cannabinoid Receptor 1

Cited by 39 publications

References 42 publications

Atypical Responsiveness of the Orphan Receptor GPR55 to Cannabinoid Ligands

Atypical Responsiveness of the Orphan Receptor GPR55 to Cannabinoid Ligands

Antagonism of Dopamine Receptor 2 Long Affects Cannabinoid Receptor 1 Signaling in a Cell Culture Model of Striatal Medium Spiny Projection Neurons

CB1 Receptor Allosteric Modulators Display Both Agonist and Signaling Pathway Specificity

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CB₁ Receptor Allosteric Modulators Display Both Agonist and Signaling Pathway Specificity