A cell-based assay to assess the persistence of action of agonists acting at recombinant human β2 adrenoceptors

Currently, there are no available approaches to cure or slow down the progression of Alzheimer’s disease (AD), which is characterized by the accumulation of extracellular amyloid-β (Aβ) deposits and intraneuronal tangles composed of hyperphosphorylated tau. β2 adrenergic receptors (β2ARs) are expressed throughout the cortex and hippocampus and play a key role in cognitive functions. Alterations in the function of these receptors have been linked to Alzheimer’s disease; however these data remain controversial as apparent contradicting reports have been published. Given the current demographics of growing elderly population and the high likelihood of concurrent beta-blocker use for other chronic conditions, more studies into the role of this receptor in AD animal models are needed. Here we show that administration of ICI 118,551, a selective β2AR antagonist, exacerbates cognitive deficits in a mouse model of AD, the 3xTg-AD mice. Neuropathologically, ICI 118,551 increased Aβ levels and Aβ plaque burden. Concomitantly, ICI 118,551-treated 3xTg-AD mice showed an increase in tau phosphorylation and accumulation. Mechanistically, these changes were linked to an increase in amyloidogenic APP processing. These results suggest that under the conditions used here, selective pharmacological inhibition of β2ARs has detrimental effects on AD-like pathology in mice. Overall, these studies strengthen the notion that the link between β2ARs and AD is likely highly complex and suggest caution in generalizing the beneficial effects of beta-blockers on AD.

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“…1A), a selective β2AR antagonist (Summerhill, et al, 2008), which was delivered via daily intraperitoneal (i.p.) injections for 6 weeks (Fig.…”

Section: Resultsmentioning

confidence: 99%

Administration of a selective β2 adrenergic receptor antagonist exacerbates neuropathology and cognitive deficits in a mouse model of Alzheimer's disease

Branca

Wisely

Hartman

et al. 2014

Neurobiology of Aging

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“…Washout Experiments-Agonist activity was compared with and without serial washings shortly after addition of ligand as previously described (14,23). Briefly, HEK293 cells were plated and transfected as described above with the addition of a poly-L-lysine plate pretreatment step to enhance cell adhesion.…”

Section: Methodsmentioning

confidence: 99%

Development of a Membrane-anchored Chemerin Receptor Agonist as a Novel Modulator of Allergic Airway Inflammation and Neuropathic Pain

Doyle

Krishnaji

Zhu

et al. 2014

Journal of Biological Chemistry

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Background:The chemerin-CMKLR1 axis modulates inflammation; ligands for CMKLR1 have short half-lives. Results: We have utilized a novel technology to develop a long-acting, high potency membrane-anchored CMKLR1 agonist. Conclusion: This ligand decreases allergic airway inflammation and neuropathic pain in mice. Significance: Our approach can be applied to develop other candidate therapeutics targeting peptide hormone receptors implicated as modulators of disease.

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“…Methodology to determine the persistence (duration of effect) and reassertion behavior (blockade of agonist receptor activation with an antagonist followed by the restoration of an agonist effect postwashout of all agents with no subsequent agonist addition) of b 2 -AR agonists were adapted from procedures previously described (Summerhill et al, 2008;Patel et al, 2011). In brief, 55,000 cells/well of b 2 -AR CHO cells were added to clear, V-bottomed, 96-well plates and then treated with one of the conditions detailed below.…”

Section: Lance Camp Assaysmentioning

confidence: 99%

In Vitro Pharmacological Characterization of Vilanterol, a Novel Long-Actingβ₂-Adrenoceptor Agonist with 24-Hour Duration of Action

Slack

Barrett

Morrison

et al. 2012

J Pharmacol Exp Ther

104

124

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Vilanterol trifenatate (vilanterol) is a novel, long-acting b 2 -adrenoceptor (b 2 -AR) agonist with 24 h activity. In this study, we describe the preclinical pharmacological profile of vilanterol using radioligand binding and cAMP studies in recombinant assays as well as human and guinea pig tissue systems to characterize b 2 -AR binding and functional properties. Vilanterol displayed a subnanomolar affinity for the b 2 -AR that was comparable with that of salmeterol but higher than olodaterol, formoterol, and indacaterol. In cAMP functional activity studies, vilanterol demonstrated similar selectivity as salmeterol for b 2 -over b 1 -AR and b 3 -AR, but a significantly improved selectivity profile than formoterol and indacaterol. Vilanterol also showed a level of intrinsic efficacy that was comparable to indacaterol but significantly greater than that of salmeterol. In cellular cAMP production and tissue-based studies measuring persistence and reassertion, vilanterol had a persistence of action comparable with indacaterol and longer than formoterol. In addition, vilanterol demonstrated reassertion activity in both cell and tissue systems that was comparable with salmeterol and indacaterol but longer than formoterol. In human airways, vilanterol was shown to have a faster onset and longer duration of action than salmeterol, exhibiting a significant level of bronchodilation 22 h after treatment. From these investigations, the data for vilanterol are consistent, showing that it is a novel, potent, and selective b 2 -AR receptor agonist with a long duration of action. This pharmacological profile combined with clinical data is consistent with once a day dosing of vilanterol in the treatment of both asthma and chronic obstructive pulmonary disease (COPD).

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A cell-based assay to assess the persistence of action of agonists acting at recombinant human β2 adrenoceptors

Cited by 31 publications

References 27 publications

Administration of a selective β2 adrenergic receptor antagonist exacerbates neuropathology and cognitive deficits in a mouse model of Alzheimer's disease

Administration of a selective β2 adrenergic receptor antagonist exacerbates neuropathology and cognitive deficits in a mouse model of Alzheimer's disease

Development of a Membrane-anchored Chemerin Receptor Agonist as a Novel Modulator of Allergic Airway Inflammation and Neuropathic Pain

In Vitro Pharmacological Characterization of Vilanterol, a Novel Long-Actingβ₂-Adrenoceptor Agonist with 24-Hour Duration of Action

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A cell-based assay to assess the persistence of action of agonists acting at recombinant human β2 adrenoceptors

Cited by 31 publications

References 27 publications

Administration of a selective β2 adrenergic receptor antagonist exacerbates neuropathology and cognitive deficits in a mouse model of Alzheimer's disease

Administration of a selective β2 adrenergic receptor antagonist exacerbates neuropathology and cognitive deficits in a mouse model of Alzheimer's disease

Development of a Membrane-anchored Chemerin Receptor Agonist as a Novel Modulator of Allergic Airway Inflammation and Neuropathic Pain

In Vitro Pharmacological Characterization of Vilanterol, a Novel Long-Actingβ2-Adrenoceptor Agonist with 24-Hour Duration of Action

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In Vitro Pharmacological Characterization of Vilanterol, a Novel Long-Actingβ₂-Adrenoceptor Agonist with 24-Hour Duration of Action