Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17

Buccioni, Michela; Marucci, Gabriella; Ben, Diego Dal; Giacobbe, Dania; Lambertucci, Catia; Soverchia, Laura; Thomas, Ajiroghene; Volpini, Rosaria; Cristalli, Gloria

doi:10.1007/s11302-011-9245-8

Cited by 44 publications

(47 citation statements)

References 11 publications

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“…Although multiple follow-up studies consistent with this original report have been published by the same group (Parravicini et al, 2008;Pugliese et al, 2009;Buccioni et al, 2011;Daniele et al, 2011;Coppi et al, 2013), Benned-Jensen and Rosenkilde (2010) reported that UDP and UDP-glucose, but not cysteinyl leukotrienes, promoted GPR17-dependent guanosine 59-O-(3-[…”

Section: Introductionsupporting

confidence: 60%

“…A recent study by a different group analyzed the pharmacological selectivity of GPR17 expressed in HEK293 cells using the GloSensor cAMP assay (Promega, Madison, WI) and observed activation by UDP, UDP-galactose, and UDP-glucose (Buccioni et al, 2011). However, the authors were unable to demonstrate inhibition of cAMP accumulation by two different immunocompetitive cAMP assays and required the increased sensitivity of the GloSensor assay to observe inhibition of adenylyl cyclase.…”

Section: Discussionmentioning

confidence: 99%

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Is GPR17 a P2Y/Leukotriene Receptor? Examination of Uracil Nucleotides, Nucleotide Sugars, and Cysteinyl Leukotrienes as Agonists of GPR17

Harden

Nicholas

2013

J Pharmacol Exp Ther

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The orphan receptor GPR17 has been reported to be activated by UDP, UDP-sugars, and cysteinyl leukotrienes, and coupled to intracellular Ca 21 mobilization and inhibition of cAMP accumulation, but other studies have reported either a different agonist profile or lack of agonist activity altogether. To determine if GPR17 is activated by uracil nucleotides and leukotrienes, the hemagglutinintagged receptor was expressed in five different cell lines and the signaling properties of the receptor were investigated. In C6, 1321N1, or Chinese hamster ovary (CHO) cells stably expressing GPR17, UDP, UDP-glucose, UDP-galactose, and cysteinyl leukotriene C4 (LTC4) all failed to promote inhibition of forskolin-stimulated cAMP accumulation, whereas both UDP and UDP-glucose promoted marked inhibition (.80%) of forskolinstimulated cAMP accumulation in C6 and CHO cells expressing the P2Y 14 receptor. Likewise, none of these compounds promoted accumulation of inositol phosphates in COS-7 or human embryonic kidney 293 cells transiently transfected with GPR17 alone or cotransfected with Ga q/i5 , which links G icoupled receptors to the G q -regulated phospholipase C (PLC) signaling pathway, or PLC«, which is activated by the Ga 12/13 signaling pathway. Moreover, none of these compounds promoted internalization of GPR17 in 1321N1-GPR17 cells. Consistent with previous reports, coexpression experiments of GPR17 with cysteinyl leukotriene receptor 1 (CysLTR1) suggested that GPR17 acts as a negative regulator of CysLTR1. Taken together, these data suggest that UDP, UDP-glucose, UDP-galactose, and LTC4 are not the cognate ligands of GPR17.

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Section: Introductionsupporting

confidence: 60%

Section: Discussionmentioning

confidence: 99%

Is GPR17 a P2Y/Leukotriene Receptor? Examination of Uracil Nucleotides, Nucleotide Sugars, and Cysteinyl Leukotrienes as Agonists of GPR17

Harden

Nicholas

2013

J Pharmacol Exp Ther

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“…Both rodent and human GPR17 dually respond to uracil nucleotides (UDP and UDP-glucose) and arachidonic acid-derived cysteinyl-leukotrienes (LTD 4 and LTE 4 ), two families of alerting "danger signals" accumulating at injury sites [12][13][14][15][16]. The purinergic component of GPR17 has been confirmed in two additional independent studies [17,18], whereas response to cysteinyl-leukotrienes has been proposed to be related to the formation of heteromers [17,[19][20][21]. In this respect, in a previous in silico and in vitro study on GPR17 ligands performed by our group [22], we proposed a possible allosteric effect of the CysLT1R antagonist montelukast on GPR17 uracil ligands, suggesting that montelukast may act on a yet uncharacterized binding site differing from the principal orthosteric one.…”

Section: Introductionmentioning

confidence: 86%

Changes of the GPR17 receptor, a new target for neurorepair, in neurons and glial cells in patients with traumatic brain injury

Franke

Parravicini

Lecca

et al. 2013

Purinergic Signalling

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Unveiling the mechanisms participating in the damage and repair of traumatic brain injury (TBI) is fundamental to develop new therapies. The P2Y-like GPR17 receptor has recently emerged as a sensor of damage and a key actor in lesion remodeling/repair in the rodent brain, but its role in humans is totally unknown. Here, we characterized GPR17 expression in brain specimens from seven intensive care unit TBI patients undergoing neurosurgery for contusion removal and from 28 autoptic TBI cases (and 10 control subjects of matched age and gender) of two university hospitals. In both neurosurgery and autoptic samples, GPR17 expression was strong inside the contused core and progressively declined distally according to a spatio-temporal gradient. Inside and around the core, GPR17 labeled dying neurons, reactive astrocytes, and activated microglia/macrophages. In peri-contused parenchyma, GPR17 decorated oligodendrocyte precursor cells (OPCs) some of which had proliferated, indicating re-myelination attempts. In autoptic cases, GPR17 expression positively correlated with death for intracranial complications and negatively correlated with patients' post-traumatic survival. Data indicate lesion-specific sequential involvement of GPR17 in the (a) death of irreversibly damaged neurons, (b) activation of microglia/macrophages remodeling the lesion, and (c) activation/proliferation of multipotent parenchymal progenitors (both reactive astrocytes and OPCs) starting repair processes. Data validate GPR17 as a target for neurorepair and are particularly relevant to setting up new therapies for TBI patients.

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“…In particular, we have utilized the non-selective GPR17 agonist UDP-glucose (UDP-glc; 100 μM [16,[20][21][22][23][31][32][33][34]) and antagonist Cangrelor (Cang; 10 μM), in parallel to VPA (500 μM). All reagents were obtained from Sigma-Aldrich, except for Cangrelor that was a kind gift of The Medicines Company (Parsippany, NJ, USA).…”

Section: Pharmacological Treatmentsmentioning

confidence: 99%

“…We chose a GPR17 antagonist (Cang; 10 μM) and a GPR17 agonist (UDP-glc; 100 μM [16,[20][21][22][23][31][32][33][34]). In neurogenic protocol #1, cells were treated with the different pharmacological agents during phase C only, whereas in neurogenic protocol #2 cells were treated during both phase DM and SCM (see Fig.…”

Section: Valproic Acid Implements the Neurogenic Potential Of Opcs Anmentioning

confidence: 99%

A new role for the P2Y-like GPR17 receptor in the modulation of multipotency of oligodendrocyte precursor cells in vitro

Boccazzi

Lecca

Marangon

et al. 2016

Purinergic Signalling

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Oligodendrocyte precursor cells (OPCs, also called NG2 cells) are scattered throughout brain parenchyma, where they function as a reservoir to replace lost or damaged oligodendrocytes, the myelin-forming cells. The hypothesis that, under some circumstances, OPCs can actually behave as multipotent cells, thus generating astrocytes and neurons as well, has arisen from some in vitro and in vivo evidence, but the molecular pathways controlling this alternative fate of OPCs are not fully understood. Their identification would open new opportunities for neuronal replace strategies, by fostering the intrinsic ability of the brain to regenerate. Here, we show that the anti-epileptic epigenetic modulator valproic acid (VPA) can promote the generation of new neurons from NG2 + OPCs under neurogenic protocols in vitro, through their initial de-differentiation to a stem cell-like phenotype that then evolves to Bhybrid^cell population, showing OPC morphology but expressing the neuronal marker βIII-tubulin and the GPR17 receptor, a key determinant in driving OPC transition towards myelinating oligodendrocytes. Under these conditions, the pharmacological blockade of the P2Y-like receptor GPR17 by cangrelor, a drug recently approved for human use, partially mimics the effects mediated by VPA thus accelerating cells' neurogenic conversion. These data show a co-localization between neuronal markers and GPR17 in vitro, and suggest that, besides its involvement in oligodendrogenesis, GPR17 can drive the fate of neural precursor cells by instructing precursors towards the neuronal lineage. Being a membrane receptor, GPR17 represents an ideal Bdruggable^target to be exploited for innovative regenerative approaches to acute and chronic brain diseases.

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Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17

Cited by 44 publications

References 11 publications

Is GPR17 a P2Y/Leukotriene Receptor? Examination of Uracil Nucleotides, Nucleotide Sugars, and Cysteinyl Leukotrienes as Agonists of GPR17

Is GPR17 a P2Y/Leukotriene Receptor? Examination of Uracil Nucleotides, Nucleotide Sugars, and Cysteinyl Leukotrienes as Agonists of GPR17

Changes of the GPR17 receptor, a new target for neurorepair, in neurons and glial cells in patients with traumatic brain injury

A new role for the P2Y-like GPR17 receptor in the modulation of multipotency of oligodendrocyte precursor cells in vitro

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