Mechanistic insights into the role of the chemokine CCL2/CCR2 axis in dorsal root ganglia to peripheral inflammation and pain hypersensitivity.
Animals
Cells, Cultured
Chemokine CCL2
/ metabolism
Freund's Adjuvant
/ toxicity
Ganglia, Spinal
/ drug effects
Hyperalgesia
/ chemically induced
Inflammation
/ chemically induced
Injections, Spinal
Male
Pain
/ chemically induced
Pyrrolidines
/ administration & dosage
Rats
Rats, Sprague-Dawley
Receptors, CCR2
/ antagonists & inhibitors
CFA
Calcium mobilization
Dorsal root ganglion
Formalin
INCB3344
MCP-1
Nociception
Nociceptor
Peripheral inflammation
Substance P
Journal
Journal of neuroinflammation
ISSN: 1742-2094
Titre abrégé: J Neuroinflammation
Pays: England
ID NLM: 101222974
Informations de publication
Date de publication:
23 Mar 2021
23 Mar 2021
Historique:
received:
16
10
2020
accepted:
05
03
2021
entrez:
24
3
2021
pubmed:
25
3
2021
medline:
18
11
2021
Statut:
epublish
Résumé
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. 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. 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. 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.
Sections du résumé
BACKGROUND
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
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
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
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.
Identifiants
pubmed: 33757529
doi: 10.1186/s12974-021-02125-y
pii: 10.1186/s12974-021-02125-y
pmc: PMC7986025
doi:
Substances chimiques
Ccl2 protein, rat
0
Ccr2 protein, rat
0
Chemokine CCL2
0
INCB3344
0
Pyrrolidines
0
Receptors, CCR2
0
Freund's Adjuvant
9007-81-2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
79Subventions
Organisme : CIHR
ID : MOP-74618
Pays : Canada
Références
Ther Adv Musculoskelet Dis. 2019 Jul 23;11:1759720X19864492
pubmed: 31384314
Bioorg Med Chem Lett. 2010 Dec 15;20(24):7473-8
pubmed: 21036044
Curr Med Chem. 2008;15(27):2866-75
pubmed: 18991641
Biology (Basel). 2020 Jul 29;9(8):
pubmed: 32751156
Neuroreport. 2008 Jan 22;19(2):183-6
pubmed: 18185105
Neurosci Res. 2004 Apr;48(4):463-9
pubmed: 15041200
Gen Pharmacol. 1998 Jan;30(1):5-11
pubmed: 9457475
J Neuroinflammation. 2012 Jul 09;9:136
pubmed: 22721162
J Neurochem. 2008 Jul;106(2):757-69
pubmed: 18419759
J Leukoc Biol. 2008 Sep;84(3):587-94
pubmed: 18467654
Ann Rheum Dis. 2006 Mar;65(3):294-300
pubmed: 16107514
Ann N Y Acad Sci. 2002 Jun;966:343-54
pubmed: 12114291
Inflamm Cell Signal. 2014;1(3):
pubmed: 26052540
Arthritis Rheum. 2008 Sep 15;59(9):1257-69
pubmed: 18759256
J Neurochem. 2007 Nov;103(4):1628-43
pubmed: 17883394
Bratisl Lek Listy. 2009;110(10):641-6
pubmed: 20017457
Brain Res Bull. 2005 Jul 15;66(1):50-8
pubmed: 15925144
Brain Res Bull. 2013 Jun;95:21-7
pubmed: 23562605
Mol Pain. 2013 Nov 08;9:57
pubmed: 24206615
J Cell Physiol. 2019 Jul;234(7):10018-10031
pubmed: 30536757
Neuropsychopharmacology. 2018 Oct;43(11):2320-2330
pubmed: 29993042
J Exp Med. 1997 Jul 7;186(1):131-7
pubmed: 9207007
Arthritis Rheumatol. 2020 Feb;72(2):220-233
pubmed: 31908163
Curr Med Chem. 2007;14(23):2456-70
pubmed: 17979699
J Neurosci Res. 2005 Oct 1;82(1):51-62
pubmed: 16047385
J Neurosci. 2011 Dec 14;31(50):18381-90
pubmed: 22171040
Mod Rheumatol. 2003 Dec;13(4):301-4
pubmed: 24387250
Prog Brain Res. 1996;113:343-59
pubmed: 9009744
Arthritis Rheum. 2001 May;44(5):1022-32
pubmed: 11352233
Ann Rheum Dis. 2005 Jan;64(1):1-2
pubmed: 15608297
Bull Exp Biol Med. 2014 Dec;158(2):192-6
pubmed: 25430645
Eur J Pain. 2009 Sep;13(8):812-9
pubmed: 18976941
Osteoarthritis Cartilage. 2017 Mar;25(3):406-412
pubmed: 27746376
J Neurochem. 2008 Jan;104(1):254-63
pubmed: 17944871
J Neurosci. 2011 Apr 13;31(15):5865-75
pubmed: 21490228
Eur J Pharmacol. 2011 Oct 1;668(1-2):163-8
pubmed: 21749865
Physiol Behav. 2011 Sep 1;104(3):495-502
pubmed: 21620878
Mol Pain. 2009 Aug 12;5:48
pubmed: 19674450
Pain. 2009 Apr;142(3):275-283
pubmed: 19233564
Cell Mol Neurobiol. 2014 Jan;34(1):143-56
pubmed: 24122510
Ann Rheum Dis. 2017 May;76(5):914-922
pubmed: 27965260
Biosci Rep. 2014 Jun 18;34(3):
pubmed: 24724624
Arch Intern Med. 2002 Feb 11;162(3):292-8
pubmed: 11822921
J Neuroimmunol. 2008 Jul 31;198(1-2):62-8
pubmed: 18538863
Eur J Pain. 2009 Mar;13(3):263-72
pubmed: 18554968
Eur J Pain. 2007 Feb;11(2):125-38
pubmed: 16682240
Arthritis Res Ther. 2007;9(3):214
pubmed: 17572915
Brain Res Rev. 2009 Apr;60(1):125-34
pubmed: 19146875
Neurosci Bull. 2018 Feb;34(1):13-21
pubmed: 28265898
Biochem Biophys Res Commun. 2009 Sep 18;387(2):251-5
pubmed: 19576173
Eur J Neurosci. 2009 May;29(9):1896-904
pubmed: 19473241
Anesthesiology. 2010 May;112(5):1250-8
pubmed: 20395830
Postgrad Med. 2020 May;132(4):377-384
pubmed: 32100608
J Cereb Blood Flow Metab. 2010 Mar;30(3):459-73
pubmed: 19904283
Pain. 1993 Jul;54(1):43-50
pubmed: 8378102
Arthritis Rheum. 2005 Dec;52(12):3718-30
pubmed: 16320322
J Rheumatol. 2001 Jan;28(1):41-6
pubmed: 11196541
Cell Mol Life Sci. 2017 Sep;74(18):3275-3291
pubmed: 28389721
Neurosci Bull. 2020 Nov;36(11):1344-1354
pubmed: 32809188
Neuroscience. 2007 Nov 9;149(3):706-14
pubmed: 17870246
Pain. 1977 Dec;4(2):161-174
pubmed: 564014
Osteoarthritis Cartilage. 2017 Jun;25(6):914-925
pubmed: 27856294
Cancer Chemother Pharmacol. 2013 Apr;71(4):1041-50
pubmed: 23385782
Int Immunopharmacol. 2020 Jul;84:106540
pubmed: 32402949
J Neurochem. 2011 Sep;118(5):680-94
pubmed: 21722132
Ann Clin Biochem. 2015 Mar;52(Pt 2):276-82
pubmed: 25005456
Br J Rheumatol. 1998 Aug;37(8):870-3
pubmed: 9734678
J Clin Invest. 1992 Sep;90(3):772-9
pubmed: 1522232
Fundam Clin Pharmacol. 2016 Jun;30(3):235-47
pubmed: 26820818
J Neuroinflammation. 2012 Aug 08;9:189
pubmed: 22870919
J Neurotrauma. 2008 May;25(5):427-48
pubmed: 18338959
Curr Rheumatol Rep. 2013 Jan;15(1):300
pubmed: 23292816
J Neurochem. 2006 Jun;97(5):1412-8
pubmed: 16696851
Neuropharmacology. 2014 Jun;81:75-84
pubmed: 24495396
Nat Rev Rheumatol. 2014 Oct;10(10):581-92
pubmed: 24861185
Nat Rev Neurosci. 2007 Nov;8(11):895-903
pubmed: 17948033
J Rheumatol. 2005 Nov;32(11):2205-11
pubmed: 16265704
J Pathol. 1997 May;182(1):106-14
pubmed: 9227349
Immunity. 2009 Nov 20;31(5):711-21
pubmed: 19836265
J Neuroimmunol. 2016 Aug 15;297:9-19
pubmed: 27397071
Nat Rev Neurosci. 2003 Jun;4(6):444-55
pubmed: 12778117
J Pain. 2009 Mar;10(3):300-5
pubmed: 19070549
FASEB J. 2013 Sep;27(9):3741-52
pubmed: 23756650
Front Biosci (Landmark Ed). 2009 Jan 01;14:540-51
pubmed: 19273084
Annu Rev Pharmacol Toxicol. 2002;42:469-99
pubmed: 11807180
Osteoarthritis Cartilage. 2006 Oct;14(10):1033-40
pubmed: 16713310
Neurosci Lett. 2012 May 23;517(1):60-3
pubmed: 22531750
J Immunol. 2005 Oct 15;175(8):5370-8
pubmed: 16210643
Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7947-52
pubmed: 12808141
Nat Rev Drug Discov. 2014 Jul;13(7):533-48
pubmed: 24948120
J Neurosci. 2009 Apr 1;29(13):4096-108
pubmed: 19339605
Mol Interv. 2009 Aug;9(4):188-95
pubmed: 19720751