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Article ; Online: Ligand-dependent intracellular trafficking of the G protein-coupled P2Y

Girard, Mélissa / Bellefeuille, Steve Dagenais / Eiselt, Émilie / Arguin, Guillaume / Longpré, Jean-Michel / Sarret, Philippe / Gendron, Fernand-Pierre

Biochimica et biophysica acta. Molecular cell research

2023 Volume 1870, Issue 5, Page(s) 119476

Abstract: Endosomal trafficking is intricately linked to G protein-coupled receptors (GPCR) fate and signaling. Extracellular uridine diphosphate (UDP) acts as a signaling molecule by selectively activating the GPCR ... ...

Abstract	Endosomal trafficking is intricately linked to G protein-coupled receptors (GPCR) fate and signaling. Extracellular uridine diphosphate (UDP) acts as a signaling molecule by selectively activating the GPCR P2Y
MeSH term(s)	Humans ; Ligands ; Receptors, G-Protein-Coupled/genetics ; Receptors, G-Protein-Coupled/metabolism ; Uridine Diphosphate/metabolism ; GTP-Binding Proteins/metabolism
Chemical Substances	purinoceptor P2Y6 ; Ligands ; Receptors, G-Protein-Coupled ; Uridine Diphosphate (58-98-0) ; GTP-Binding Proteins (EC 3.6.1.-)
Language	English
Publishing date	2023-04-12
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	60-7
ISSN	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbamcr.2023.119476
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Article ; Online: Assessing Gα

Besserer-Offroy, Élie / Brouillette, Rebecca L / Longpré, Jean-Michel / Sarret, Philippe

Bio-protocol

2020 Volume 10, Issue 16, Page(s) e3715

Abstract: Cell-based functional assays are an important part of compound screening and drug lead optimization, and they can also play a crucial role in the determination of the residues involved in ligand binding and signaling for a particular G-protein-coupled ... ...

Abstract	Cell-based functional assays are an important part of compound screening and drug lead optimization, and they can also play a crucial role in the determination of the residues involved in ligand binding and signaling for a particular G-protein-coupled receptor. Conventional methods used for Gα
Language	English
Publishing date	2020-08-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2833269-6
ISSN	2331-8325 ; 2331-8325
ISSN (online)	2331-8325
ISSN	2331-8325
DOI	10.21769/BioProtoc.3715
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Article ; Online: Mechanistic insights into the role of the chemokine CCL2/CCR2 axis in dorsal root ganglia to peripheral inflammation and pain hypersensitivity.

Dansereau, Marc-André / Midavaine, Élora / Bégin-Lavallée, Valérie / Belkouch, Mounir / Beaudet, Nicolas / Longpré, Jean-Michel / Mélik-Parsadaniantz, Stéphane / Sarret, Philippe

Journal of neuroinflammation

2021 Volume 18, Issue 1, Page(s) 79

Abstract: Background: Pain is reported as the leading cause of disability in the common forms of inflammatory arthritis conditions. Acting as a key player in nociceptive processing, neuroinflammation, and neuron-glia communication, the chemokine CCL2/CCR2 axis ... ...

Abstract	Background: Pain is reported as the leading cause of disability in the common forms of inflammatory arthritis conditions. Acting as a key player in nociceptive processing, neuroinflammation, and neuron-glia communication, the chemokine CCL2/CCR2 axis holds great promise for controlling chronic painful arthritis. Here, we investigated how the CCL2/CCR2 system in the dorsal root ganglion (DRG) contributes to the peripheral inflammatory pain sensitization. Methods: Repeated intrathecal (i.t.) administration of the CCR2 antagonist, INCB3344 was tested for its ability to reverse the nociceptive-related behaviors in the tonic formalin and complete Freund's adjuvant (CFA) inflammatory models. We further determined by qPCR the expression of CCL2/CCR2, SP and CGRP in DRG neurons from CFA-treated rats. Using DRG explants, acutely dissociated primary sensory neurons and calcium mobilization assay, we also assessed the release of CCL2 and sensitization of nociceptors. Finally, we examined by immunohistochemistry following nerve ligation the axonal transport of CCL2, SP, and CGRP from the sciatic nerve of CFA-treated rats. Results: We first found that CFA-induced paw edema provoked an increase in CCL2/CCR2 and SP expression in ipsilateral DRGs, which was decreased after INCB3344 treatment. This upregulation in pronociceptive neuromodulators was accompanied by an enhanced nociceptive neuron excitability on days 3 and 10 post-CFA, as revealed by the CCR2-dependent increase in intracellular calcium mobilization following CCL2 stimulation. In DRG explants, we further demonstrated that the release of CCL2 was increased following peripheral inflammation. Finally, the excitation of nociceptors following peripheral inflammation stimulated the anterograde transport of SP at their peripheral nerve terminals. Importantly, blockade of CCR2 reduced sensory neuron excitability by limiting the calcium mobilization and subsequently decreased peripheral transport of SP towards the periphery. Finally, pharmacological inhibition of CCR2 reversed the pronociceptive action of CCL2 in rats receiving formalin injection and significantly reduced the neurogenic inflammation as well as the stimuli-evoked and movement-evoked nociceptive behaviors in CFA-treated rats. Conclusions: Our results provide significant mechanistic insights into the role of CCL2/CCR2 within the DRG in the development of peripheral inflammation, nociceptor sensitization, and pain hypersensitivity. We further unveil the therapeutic potential of targeting CCR2 for the treatment of painful inflammatory disorders.
MeSH term(s)	Animals ; Cells, Cultured ; Chemokine CCL2/metabolism ; Freund's Adjuvant/toxicity ; Ganglia, Spinal/drug effects ; Ganglia, Spinal/metabolism ; Hyperalgesia/chemically induced ; Hyperalgesia/drug therapy ; Hyperalgesia/metabolism ; Inflammation/chemically induced ; Inflammation/drug therapy ; Inflammation/metabolism ; Injections, Spinal ; Male ; Pain/chemically induced ; Pain/drug therapy ; Pain/metabolism ; Pyrrolidines/administration & dosage ; Rats ; Rats, Sprague-Dawley ; Receptors, CCR2/antagonists & inhibitors ; Receptors, CCR2/metabolism
Chemical Substances	Ccl2 protein, rat ; Ccr2 protein, rat ; Chemokine CCL2 ; INCB3344 ; Pyrrolidines ; Receptors, CCR2 ; Freund's Adjuvant (9007-81-2)
Language	English
Publishing date	2021-03-23
Publishing country	England
Document type	Journal Article
ISSN	1742-2094
ISSN (online)	1742-2094
DOI	10.1186/s12974-021-02125-y
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Article ; Online: Design, Structural Optimization, and Characterization of the First Selective Macrocyclic Neurotensin Receptor Type 2 Non-opioid Analgesic.

Chartier, Magali / Desgagné, Michael / Sousbie, Marc / Côté, Jérôme / Longpré, Jean-Michel / Marsault, Eric / Sarret, Philippe

Journal of medicinal chemistry

2021 Volume 64, Issue 4, Page(s) 2110–2124

Abstract: Neurotensin (NT) receptor type 2 (NTS2) represents an attractive target for the development of new NT-based analgesics. Here, we report the synthesis and ... ...

Abstract	Neurotensin (NT) receptor type 2 (NTS2) represents an attractive target for the development of new NT-based analgesics. Here, we report the synthesis and functional
MeSH term(s)	Analgesics/chemical synthesis ; Analgesics/therapeutic use ; Animals ; Drug Design ; Male ; Molecular Structure ; Neurotensin/analogs & derivatives ; Neurotensin/therapeutic use ; Pain/drug therapy ; Peptide Fragments/chemical synthesis ; Peptide Fragments/therapeutic use ; Peptides, Cyclic/chemical synthesis ; Peptides, Cyclic/therapeutic use ; Rats, Sprague-Dawley ; Receptors, Neurotensin/metabolism ; Structure-Activity Relationship ; Rats
Chemical Substances	Analgesics ; Ntsr2 protein, rat ; Peptide Fragments ; Peptides, Cyclic ; Receptors, Neurotensin ; Neurotensin (39379-15-2)
Language	English
Publishing date	2021-02-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218133-2
ISSN	1520-4804 ; 0022-2623
ISSN (online)	1520-4804
ISSN	0022-2623
DOI	10.1021/acs.jmedchem.0c01726
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Article ; Online: The combination of opioid and neurotensin receptor agonists improves their analgesic/adverse effect ratio.

Eiselt, Emilie / Côté, Jérôme / Longpré, Jean-Michel / Blais, Véronique / Sarret, Philippe / Gendron, Louis

European journal of pharmacology

2019 Volume 848, Page(s) 80–87

Abstract: Opioid and neurotensin (NT) receptors are expressed in both central and peripheral nervous systems where they modulate nociceptive responses. Nowadays, opioid analgesics like morphine remain the most prescribed drugs for the treatment of moderate to ... ...

Abstract	Opioid and neurotensin (NT) receptors are expressed in both central and peripheral nervous systems where they modulate nociceptive responses. Nowadays, opioid analgesics like morphine remain the most prescribed drugs for the treatment of moderate to severe pain. However, despite their daily used, opioids can produce life-threatening side effects, such as constipation or respiratory depression. Besides, NT analogs exert strong opioid-independent analgesia. Here, we thus hypothesized that the combined use of opioid and NT agonists would require lower doses to produce significant analgesic effects, hence decreasing opioid-induced adverse effects. We used isobologram analyses to determine if the combination of a NT brain-penetrant analog, An2-NT(8-13) with morphine results in an inhibitory, synergistic or additive analgesic response. We found that intravenous administration of An2-NT(8-13) reduced by 90% the nocifensive behaviors induced by formalin injection, at the dose of 0.018 mg/kg. Likewise, subcutaneous morphine reduced pain by 90% at 1.8 mg/kg. Importantly, isobologram analyses revealed that the co-injection of An2-NT(8-13) with morphine induced an additive analgesic response. We finally assessed the effects of morphine and An2-NT(8-13) on the gastrointestinal tract motility using the charcoal meal test. As opposed to morphine which significantly reduced the intestinal motility at the analgesic effective dose of 1.8 mg/kg, An2-NT(8-13) did not affect the charcoal meal intestinal transit at 0.018 mg/kg. Interestingly, at the dose providing 90% pain relief, the co-administration of morphine with An2-NT(8-13) had a reduced effect on constipation. Altogether, these results suggest that combining NT agonists with morphine may improve its analgesic/adverse effect ratio.
MeSH term(s)	Analgesics, Opioid/administration & dosage ; Animals ; Dose-Response Relationship, Drug ; Drug Therapy, Combination ; Male ; Morphine/administration & dosage ; Neurotensin/administration & dosage ; Pain Measurement/drug effects ; Pain Measurement/methods ; Peptide Fragments/administration & dosage ; Peptides/administration & dosage ; Rats ; Rats, Sprague-Dawley ; Receptors, Neurotensin/agonists ; Receptors, Neurotensin/metabolism ; Receptors, Opioid, mu/agonists ; Receptors, Opioid, mu/metabolism
Chemical Substances	Analgesics, Opioid ; Angiopep-2 ; Peptide Fragments ; Peptides ; Receptors, Neurotensin ; Receptors, Opioid, mu ; Neurotensin (39379-15-2) ; neurotensin (8-13) (60482-95-3) ; Morphine (76I7G6D29C)
Language	English
Publishing date	2019-01-29
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	80121-5
ISSN	1879-0712 ; 0014-2999
ISSN (online)	1879-0712
ISSN	0014-2999
DOI	10.1016/j.ejphar.2019.01.048
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Article ; Online: Signaling Modulation via Minimal C-Terminal Modifications of Apelin-13.

Théroux, Léa / Van Den Hauwe, Robin / Trân, Kien / Fournier, Justin / Desgagné, Michael / Meneboo, Nathan / Lavallée, Alexis / Fröhlich, Ulrike / Côté, Jérôme / Hollanders, Charlie / Longpré, Jean-Michel / Murza, Alexandre / Marsault, Eric / Sarret, Philippe / Boudreault, Pierre-Luc / Ballet, Steven

ACS pharmacology & translational science

2023 Volume 6, Issue 2, Page(s) 290–305

Abstract: Apelin is an endogenous peptide that is involved in many diseases such as cardiovascular diseases, obesity, and cancer, which has made it an attractive target for drug discovery. Herein, we explore the penultimate and final sequence positions of [ ... ...

Abstract	Apelin is an endogenous peptide that is involved in many diseases such as cardiovascular diseases, obesity, and cancer, which has made it an attractive target for drug discovery. Herein, we explore the penultimate and final sequence positions of [Pyr
Language	English
Publishing date	2023-01-26
Publishing country	United States
Document type	Journal Article
ISSN	2575-9108
ISSN (online)	2575-9108
DOI	10.1021/acsptsci.2c00219
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Article: Insightful Backbone Modifications Preventing Proteolytic Degradation of Neurotensin Analogs Improve NT

Previti, Santo / Vivancos, Mélanie / Rémond, Emmanuelle / Beaulieu, Sabrina / Longpré, Jean-Michel / Ballet, Steven / Sarret, Philippe / Cavelier, Florine

Frontiers in chemistry

2020 Volume 8, Page(s) 406

Abstract: Therapeutic hypothermia represents a brain-protective strategy for multiple emergency situations, such as stroke or traumatic injury. Neurotensin (NT), which exerts its effects through activation of two G protein-coupled receptors, namely NTS1 and NTS2, ... ...

Abstract	Therapeutic hypothermia represents a brain-protective strategy for multiple emergency situations, such as stroke or traumatic injury. Neurotensin (NT), which exerts its effects through activation of two G protein-coupled receptors, namely NTS1 and NTS2, induces a strong and long-lasting decrease in core body temperature after its central administration. Growing evidence demonstrates that NTS1 is the receptor subtype mediating the hypothermic action of NT. As such, potent NTS1 agonists designed on the basis of the minimal C-terminal NT(8-13) bioactive fragment have been shown to produce mild hypothermia and exert neuroprotective effects under various clinically relevant conditions. The high susceptibility of NT(8-13) to protease degradation (half-life <2 min) represents, however, a serious limitation for its use in pharmacological therapy. In light of this, we report here a structure-activity relationship study in which pairs of NT(8-13) analogs have been developed, based on the incorporation of a reduced Lys
Language	English
Publishing date	2020-06-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2711776-5
ISSN	2296-2646
ISSN	2296-2646
DOI	10.3389/fchem.2020.00406
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Article ; Online: The P2Y

Girard, Mélissa / Dagenais Bellefeuille, Steve / Eiselt, Émilie / Brouillette, Rebecca / Placet, Morgane / Arguin, Guillaume / Longpré, Jean-Michel / Sarret, Philippe / Gendron, Fernand-Pierre

Journal of cellular physiology

2020 Volume 235, Issue 12, Page(s) 9676–9690

Abstract: Cell migration is a ubiquitous process necessary to maintain and restore tissue functions. However, in cancer, cell migration leads to metastasis development and thus worsens the prognosis. Although the mechanism of cell migration is well understood, the ...

Abstract	Cell migration is a ubiquitous process necessary to maintain and restore tissue functions. However, in cancer, cell migration leads to metastasis development and thus worsens the prognosis. Although the mechanism of cell migration is well understood, the identification of new targets modulating cell migration and deciphering their signaling events could lead to new therapies to restore tissue functions in diseases, such as inflammatory bowel disease, or to block metastatic development in different forms of cancer. Previous research has identified the G-protein-coupled P2Y
MeSH term(s)	A549 Cells ; Actins/genetics ; Caco-2 Cells ; Calcium/metabolism ; Cell Movement/genetics ; Cell Surface Extensions/genetics ; Colorectal Neoplasms/genetics ; Colorectal Neoplasms/pathology ; Epithelial Cells/metabolism ; GTP-Binding Protein alpha Subunits, G12-G13/genetics ; GTP-Binding Protein alpha Subunits, Gq-G11/genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Protein Kinase C-alpha/genetics ; Receptors, G-Protein-Coupled/genetics ; Receptors, Purinergic P2/genetics ; Signal Transduction/genetics ; rho-Associated Kinases/genetics
Chemical Substances	Actins ; Receptors, G-Protein-Coupled ; Receptors, Purinergic P2 ; purinoceptor P2Y6 ; rho-Associated Kinases (EC 2.7.11.1) ; PRKCA protein, human (EC 2.7.11.13) ; Protein Kinase C-alpha (EC 2.7.11.13) ; GTP-Binding Protein alpha Subunits, G12-G13 (EC 3.6.5.1) ; GTP-Binding Protein alpha Subunits, Gq-G11 (EC 3.6.5.1) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2020-05-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3116-1
ISSN	1097-4652 ; 0021-9541
ISSN (online)	1097-4652
ISSN	0021-9541
DOI	10.1002/jcp.29779
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Article ; Online: Size-Reduced Macrocyclic Analogues of [Pyr

Tran, Kien / Sainsily, Xavier / Côté, Jérôme / Coquerel, David / Couvineau, Pierre / Saibi, Sabrina / Haroune, Lounès / Besserer-Offroy, Élie / Flynn-Robitaille, Joël / Resua Rojas, Martin / Murza, Alexandre / Longpré, Jean-Michel / Auger-Messier, Mannix / Lesur, Olivier / Bouvier, Michel / Marsault, Éric / Boudreault, Pierre-Luc / Sarret, Philippe

Journal of medicinal chemistry

2022 Volume 65, Issue 1, Page(s) 531–551

Abstract: We previously reported a series of macrocyclic analogues of [ ... ...

Abstract	We previously reported a series of macrocyclic analogues of [Pyr
MeSH term(s)	Apelin/analogs & derivatives ; Apelin/pharmacokinetics ; Apelin/pharmacology ; Apelin Receptors/drug effects ; Arrestin/drug effects ; GTP-Binding Protein alpha Subunits, G12-G13/drug effects ; HEK293 Cells ; Half-Life ; Heart/drug effects ; Humans ; Injections, Subcutaneous ; Macrocyclic Compounds/chemical synthesis ; Macrocyclic Compounds/pharmacology ; Molecular Weight ; Signal Transduction/drug effects
Chemical Substances	Apelin ; Apelin Receptors ; Arrestin ; Macrocyclic Compounds ; GTP-Binding Protein alpha Subunits, G12-G13 (EC 3.6.5.1)
Language	English
Publishing date	2022-01-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218133-2
ISSN	1520-4804 ; 0022-2623
ISSN (online)	1520-4804
ISSN	0022-2623
DOI	10.1021/acs.jmedchem.1c01708
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Article ; Online: Structural insight into apelin receptor-G protein stoichiometry.

Yue, Yang / Liu, Lier / Wu, Li-Jie / Wu, Yiran / Wang, Ling / Li, Fei / Liu, Junlin / Han, Gye-Won / Chen, Bo / Lin, Xi / Brouillette, Rebecca L / Breault, Émile / Longpré, Jean-Michel / Shi, Songting / Lei, Hui / Sarret, Philippe / Stevens, Raymond C / Hanson, Michael A / Xu, Fei

Nature structural & molecular biology

2022 Volume 29, Issue 7, Page(s) 688–697

Abstract: The technique of cryogenic-electron microscopy (cryo-EM) has revolutionized the field of membrane protein structure and function with a focus on the dominantly observed molecular species. This report describes the structural characterization of a fully ... ...

Abstract	The technique of cryogenic-electron microscopy (cryo-EM) has revolutionized the field of membrane protein structure and function with a focus on the dominantly observed molecular species. This report describes the structural characterization of a fully active human apelin receptor (APJR) complexed with heterotrimeric G protein observed in both 2:1 and 1:1 stoichiometric ratios. We use cryo-EM single-particle analysis to determine the structural details of both species from the same sample preparation. Protein preparations, in the presence of the endogenous peptide ligand ELA or a synthetic small molecule, both demonstrate these mixed stoichiometric states. Structural differences in G protein engagement between dimeric and monomeric APJR suggest a role for the stoichiometry of G protein-coupled receptor- (GPCR-)G protein coupling on downstream signaling and receptor pharmacology. Furthermore, a small, hydrophobic dimer interface provides a starting framework for additional class A GPCR dimerization studies. Together, these findings uncover a mechanism of versatile regulation through oligomerization by which GPCRs can modulate their signaling.
MeSH term(s)	Apelin Receptors/chemistry ; Apelin Receptors/metabolism ; Carrier Proteins/metabolism ; GTP-Binding Proteins/metabolism ; Humans ; Receptors, G-Protein-Coupled/chemistry ; Signal Transduction
Chemical Substances	Apelin Receptors ; Carrier Proteins ; Receptors, G-Protein-Coupled ; GTP-Binding Proteins (EC 3.6.1.-)
Language	English
Publishing date	2022-07-11
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2126708-X
ISSN	1545-9985 ; 1545-9993
ISSN (online)	1545-9985
ISSN	1545-9993
DOI	10.1038/s41594-022-00797-5
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