PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome

Kroeze, Wesley K.; Sassano, Maria F.; Huang, Xi Ping; Lansu, Katherine; McCorvy, John D.; Giguère, Patrick M.; Sciaky, Noah; Roth, Bryan L.

doi:10.1038/nsmb.3014

Cited by 618 publications

(808 citation statements)

References 51 publications

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“…Recently, new cell-based screens that measure alternative parameters and enable high-throughput analysis have been reported (Schröder et al, 2010;Inoue et al, 2012;Kroeze et al, 2015). The detection of Ca 21 is performed with Ca 21 -sensitive fluorescent probes, enabling real-time analysis.…”

Section: Introductionmentioning

confidence: 99%

Quantitative Single-Cell Analysis of Signaling Pathways Activated Immediately Downstream of Histamine Receptor Subtypes

et al. 2016

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Genetically encoded biosensors based on Förster resonance energy transfer (FRET) can visualize responses of individual cells in real time. Here, we evaluated whether FRET-based biosensors provide sufficient contrast and specificity to measure activity of G-protein-coupled receptors. The four histamine receptor subtypes (H 1 R, H 2 R, H 3 R, and H 4 R) respond to the ligand histamine by activating three canonical heterotrimeric G-protein-mediated signaling pathways with a reported high degree of specificity. Using FRET-based biosensors, we demonstrate that H 1 R activates Gaq. We also observed that H 1 R activates Gai, albeit at a 10-fold lower potency. In addition to increasing cAMP levels, most likely via Gas, we found that the H 2 R induces Gaq-mediated calcium release. The H 3 R and H 4 R activated Gai with high specificity and a high potency. We demonstrate that a number of FRET sensors provide sufficient contrast to: 1) analyze the specificity of the histamine receptor subtypes for different heterotrimeric G-protein families with single-cell resolution, 2) probe for antagonist specificity, and 3) allow the measurement of single-cell concentration-response curves.

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Section: Introductionmentioning

confidence: 99%

Quantitative Single-Cell Analysis of Signaling Pathways Activated Immediately Downstream of Histamine Receptor Subtypes

et al. 2016

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“…In the literature, EPEA and DHEA are reported to be weak agonists of the cannabinoid receptors (10). To explore if oxygenation of EPEA or DHEA by CYPs led to the formation of metabolites with altered cannabinoid receptor activity, we performed receptor functional assays using a β-arrestin recruitment PRESTO-Tango assay (42).…”

Section: Resultsmentioning

confidence: 99%

Anti-inflammatory ω-3 endocannabinoid epoxides

McDougle

Watson

Abdeen

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

149

163

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Clinical studies suggest that diets rich in ω-3 polyunsaturated fatty acids (PUFAs) provide beneficial anti-inflammatory effects, in part through their conversion to bioactive metabolites. Here we report on the endogenous production of a previously unknown class of ω-3 PUFA–derived lipid metabolites that originate from the crosstalk between endocannabinoid and cytochrome P450 (CYP) epoxygenase metabolic pathways. The ω-3 endocannabinoid epoxides are derived from docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) to form epoxyeicosatetraenoic acid-ethanolamide (EEQ-EA) and epoxydocosapentaenoic acid-ethanolamide (EDP-EA), respectively. Both EEQ-EAs and EDP-EAs are endogenously present in rat brain and peripheral organs as determined via targeted lipidomics methods. These metabolites were directly produced by direct epoxygenation of the ω-3 endocannabinoids, docosahexanoyl ethanolamide (DHEA) and eicosapentaenoyl ethanolamide (EPEA) by activated BV-2 microglial cells, and by human CYP2J2. Neuroinflammation studies revealed that the terminal epoxides 17,18-EEQ-EA and 19,20-EDP-EA dose-dependently abated proinflammatory IL-6 cytokines while increasing anti-inflammatory IL-10 cytokines, in part through cannabinoid receptor-2 activation. Furthermore the ω-3 endocannabinoid epoxides 17,18-EEQ-EA and 19,20-EDP-EA exerted antiangiogenic effects in human microvascular endothelial cells (HMVEC) and vasodilatory actions on bovine coronary arteries and reciprocally regulated platelet aggregation in washed human platelets. Taken together, the ω-3 endocannabinoid epoxides’ physiological effects are mediated through both endocannabinoid and epoxyeicosanoid signaling pathways. In summary, the ω-3 endocannabinoid epoxides are found at concentrations comparable to those of other endocannabinoids and are expected to play critical roles during inflammation in vivo; thus their identification may aid in the development of therapeutics for neuroinflammatory and cerebrovascular diseases.

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“…The following libraries were screened in quadruplicate using a modified TANGO b-arrestin recruitment assay (Barnea et al, 2008;Kroeze et al, 2015) for activity at GPR39: NCC-1 (446 compounds; National Clinical Collection), NIMH X-901 (271 compounds; National Institute of Health Clinical Collection, Bethesda, MD), Prestwick (1120 compounds; Illkirch, France), Tocris (1280 compounds; Bristol, UK), Selleckchem (1836 compounds; Munich, Germany), and Spectrum (2400 compounds; MicroSource, Gaylordsville, CT) at a drug concentration of 10 mM to which transfected cells was exposed, following the previously published method . Daughter libraries were prepared and kept at 220°C until use.…”

Section: Methodsmentioning

confidence: 99%

Discovery and Characterization of Novel GPR39 Agonists Allosterically Modulated by Zinc

et al. 2016

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In this study, we identified two previously described kinase inhibitors-3-(4-chloro-2-fluorobenzyl)-2-methyl-N-(3-methyl-1H-pyrazol-agonists by unbiased small-molecule-based screening using a b-arrestin recruitment screening approach (PRESTO-Tango). We characterized the signaling of LY2784544 and GSK2636771 and compared their signaling patterns with a previously described "GPR39-selective"at both canonical and noncanonical signaling pathways. Unexpectedly, all three compounds displayed probe-dependent and pathway-dependent allosteric modulation by concentrations of zinc reported to be physiologic. LY2784544 and GS2636771 at GPR39 in the presence of zinc were generally as potent or more potent than their reported activities against kinases in whole-cell assays. These findings reveal an unexpected role of zinc as an allosteric potentiator of small-molecule-induced activation of GPR39 and expand the list of potential kinase off-targets to include understudied G protein-coupled receptors.

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PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome

Cited by 618 publications

References 51 publications

Quantitative Single-Cell Analysis of Signaling Pathways Activated Immediately Downstream of Histamine Receptor Subtypes

Quantitative Single-Cell Analysis of Signaling Pathways Activated Immediately Downstream of Histamine Receptor Subtypes

Anti-inflammatory ω-3 endocannabinoid epoxides

Discovery and Characterization of Novel GPR39 Agonists Allosterically Modulated by Zinc

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