Double life: How GRK2 and β-arrestin signaling participate in diseases

Zhai, Ruxu; Snyder, Jonathan; Montgomery, Sarah; Sato, Priscila Y.

doi:10.1016/j.cellsig.2022.110333

Cited by 11 publications

(11 citation statements)

References 199 publications

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“…Using a β-arrestin 2 sensor, no β-arrestin 2 recruitment was detected upon agonist treatment, with neither relaxin H2 nor the SM agonists tested (Table ; Figure G). To enhance β-arrestin recruitment, G protein-coupled receptor kinase 2 (GRK2) was coexpressed with RXFP1. However, also after coexpression of GRK2 no agonist-stimulated β-arrestin 2 recruitment was observed (Figure H).…”

Section: Resultsmentioning

confidence: 99%

Discovery of Clinical Candidate AZD5462, a Selective Oral Allosteric RXFP1 Agonist for Treatment of Heart Failure

Granberg,

Sakamaki,

Larsson

et al. 2024

J. Med. Chem.

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Optimization of the highly potent and selective, yet metabolically unstable and poorly soluble hRXFP1 agonist AZ7976 led to the identification of the clinical candidate, AZD5462. Assessment of RXFP1-dependent cell signaling demonstrated that AZD5462 activates a highly similar panel of downstream pathways as relaxin H2 but does not modulate relaxin H2-mediated cAMP second messenger responsiveness. The therapeutic potential of AZD5462 was assessed in a translatable cynomolgus monkey heart failure model. Following 8 weeks of treatment with AZD5462, robust improvements in functional cardiac parameters including LVEF were observed at weeks 9, 13, and 17 without changes in heart rate or mean arterial blood pressure. AZD5462 was well tolerated in both rat and cynomolgus monkey and has successfully completed phase I studies in healthy volunteers. In summary, AZD5462 is a small molecule pharmacological mimetic of relaxin H2 signaling at RXFP1 and holds promise as a potential therapeutic approach to treat heart failure patients.

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Section: Resultsmentioning

confidence: 99%

Discovery of Clinical Candidate AZD5462, a Selective Oral Allosteric RXFP1 Agonist for Treatment of Heart Failure

Granberg,

Sakamaki,

Larsson

et al. 2024

J. Med. Chem.

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“…Recruitment of β-arrestins has also become a focus of scientific investigation in recent years because of the tremendous increase in our understanding of the importance of the interplay between the various signaling pathways of GPCRs in the development of diseases, as well as in the efficacy or side effect profile of drugs ( Sharma and Parameswaran, 2015 ; Bagnato and Rosanò, 2019 ; Bond et al, 2019 ; Zhai et al, 2022 ). We have already seen the emergence of the first drugs to exploit of preferred activation of one of several possible signal transduction pathways, also called biased signaling, such as the cardiac drug carvedilol ( Wisler et al, 2007 ; Ibrahim and Kurose, 2012 ; Kolb et al, 2022 ; Kenakin 2019 ).…”

Section: Discussionmentioning

confidence: 99%

Investigation of adenosine A1 receptor-mediated β-arrestin 2 recruitment using a split-luciferase assay

2023

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Background: Adenosine A1 receptor (A1AR) plays a prominent role in neurological and cardiac diseases and inflammatory processes. Its endogenous ligand adenosine is known to be one of the key players in the sleep–wake cycle. Like other G protein-coupled receptors (GPCRs), stimulation of A1AR leads to the recruitment of arrestins in addition to the activation of G proteins. So far, little is known about the role of these proteins in signal transduction and regulation of A1AR compared to the activation of G proteins. In this work, we characterized a live cell assay for A1AR-mediated β-arrestin 2 recruitment. We have applied this assay to a set of different compounds that interact with this receptor.Methods: Based on NanoBit® technology, a protein complementation assay was developed in which the A1AR is coupled to the large part of the nanoluciferase (LgBiT), whereas its small part (SmBiT) is fused to the N-terminus of β-arrestin 2. Stimulation of A1AR results in the recruitment of β-arrestin 2 and subsequent complementation of a functional nanoluciferase. For comparison, corresponding data on the effect of receptor stimulation on intracellular cAMP levels were collected for some data sets using the GloSensor™ assay.Results: The assay gives highly reproducible results with a very good signal-to-noise ratio. Capadenoson, in contrast to adenosine, CPA, or NECA, shows only partial agonism in this assay with respect to the recruitment of β-arrestin 2, whereas it shows full agonism in the case of the inhibitory effect of A1AR on cAMP production. By using a GRK2 inhibitor, it becomes clear that the recruitment is at least partially dependent on the phosphorylation of the receptor by this kinase. Interestingly, this was also the first time that we demonstrate the A1AR-mediated recruitment of β-arrestin 2 by stimulation with a valerian extract.Conclusion: The presented assay is a useful tool for the quantitative study of A1AR-mediated β-arrestin 2 recruitment. It allows data collection for stimulatory, inhibitory, and modulatory substances and is also suitable for more complex substance mixtures such as valerian extract.

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“…Recruitment of β-arrestins has also become a focus of scientific investigation in recent years because of the tremendous increase in our understanding of the importance of the interplay between the various signaling pathways of GPCRs in the development of diseases, as well as in the efficacy or side effect profile of drugs (Sharma and Parameswaran, 2015; Bagnato and Rosanò, 2019; Bond et al, 2019; Zhai et al, 2022). There are already first drugs that exploit the effect of preferred activation of one of several possible signal transduction pathways also called biased signaling, such as the cardiac drug carvedilol (Wisler 2007; Ibrahim 2012; Kolb 2022; Kenakin 2019).…”

Section: Discussionmentioning

confidence: 99%

Investigation of adenosine A1 receptor mediated β-arrestin 2 recruitment using a split-luciferase assay

Saecker

Häberlein

Franken

2023

Preprint

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Background: Adenosine A1 receptor (A1AR) plays a prominent role in neurological and cardiac diseases as well as during inflammatory processes. Its endogenous ligand adenosine is known to be one of the key players in the sleep-wake cycle. Like other G protein-coupled receptors, stimulation of A1AR leads to the recruitment of arrestins in addition to the activation of G proteins. So far, little is known about the role of these proteins for signal transduction and regulation of A1AR compared to the activation of G proteins. In this work, we characterized a live cell assay for the A1AR mediated β-arrestin 2 recruitment. We have applied this assay to a set of different compounds that interact with this receptor. Methods: Based on NanoBit® technology, a protein complementation assay was developed in which the A1AR is coupled to the large part of the nanoluciferase (LgBiT) whereas its small part (SmBiT) is fused to the N-terminus of β-arrestin 2. Stimulation of A1AR results in the recruitment of β-arrestin 2 and subsequent complementation of a functional nanoluciferase. For comparison, corresponding data on the effect of receptor stimulation on intracellular cAMP levels were collected for some data sets using the GloSensor™ assay. Results: The assay gives highly reproducible results with a very good signal to noise ratio. Capadenoson, in contrast to adenosine, CPA or NECA, shows only partial agonism in this assay with respect to the recruitment of β-arrestin 2, whereas it is a full agonist in the case of the inhibitory effect of A1AR on cAMP production. By using a GRK2 inhibitor, it becomes clear that the recruitment is at least partially dependent on the phosphorylation of the receptor by this kinase. Interestingly, this was also the first time that we were able to demonstrate the A1AR mediated recruitment of β-arrestin 2 by stimulation with a valerian extract. Conclusion: The presented assay is a useful tool for the quantitative study of the A1AR mediated β-arrestin 2 recruitment. It allows data collection for stimulatory, inhibitory as well as modulatory substances and is also suitable for more complex substance mixtures such as valerian extract.

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Double life: How GRK2 and β-arrestin signaling participate in diseases

Cited by 11 publications

References 199 publications

Discovery of Clinical Candidate AZD5462, a Selective Oral Allosteric RXFP1 Agonist for Treatment of Heart Failure

Discovery of Clinical Candidate AZD5462, a Selective Oral Allosteric RXFP1 Agonist for Treatment of Heart Failure

Investigation of adenosine A1 receptor-mediated β-arrestin 2 recruitment using a split-luciferase assay

Investigation of adenosine A1 receptor mediated β-arrestin 2 recruitment using a split-luciferase assay

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