Attenuation of mechanical pain hypersensitivity by treatment with Peptide5, a connexin-43 mimetic peptide, involves inhibition of NLRP3 inflammasome in nerve-injured mice

Tonkin, RS; Bowles, C; Perera, CJ; Keating, BA; Makker, PGS; Duffy, SS; Lees, JG; Tran, C; Don, AS; Fath, T; Liu, L; O'Carroll, SJ; Nicholson, LFB; Green, CR; Gorrie, C; Moalem-Taylor, G

Attenuation of mechanical pain hypersensitivity by treatment with Peptide5, a connexin-43 mimetic peptide, involves inhibition of NLRP3 inflammasome in nerve-injured mice

Tonkin, RS Bowles, C Perera, CJ Keating, BA Makker, PGS Duffy, SS Lees, JG Tran, C Don, AS Fath, T Liu, L O'Carroll, SJ Nicholson, LFB Green, CR Gorrie, C

Moalem-Taylor, G

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Publication Type:: Journal Article
Citation:: Experimental Neurology, 2018, 300 pp. 1 - 12
Issue Date:: 2018-02-01

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Field	Value	Language
dc.contributor.author	Tonkin, RS	en_US
dc.contributor.author	Bowles, C	en_US
dc.contributor.author	Perera, CJ	en_US
dc.contributor.author	Keating, BA	en_US
dc.contributor.author	Makker, PGS	en_US
dc.contributor.author	Duffy, SS	en_US
dc.contributor.author	Lees, JG	en_US
dc.contributor.author	Tran, C	en_US
dc.contributor.author	Don, AS	en_US
dc.contributor.author	Fath, T	en_US
dc.contributor.author	Liu, L	en_US
dc.contributor.author	O'Carroll, SJ	en_US
dc.contributor.author	Nicholson, LFB	en_US
dc.contributor.author	Green, CR	en_US
dc.contributor.author	Gorrie, C https://orcid.org/0000-0001-5934-2492	en_US
dc.contributor.author	Moalem-Taylor, G	en_US
dc.date.available	2017-10-17	en_US
dc.date.issued	2018-02-01	en_US
dc.identifier.citation	Experimental Neurology, 2018, 300 pp. 1 - 12	en_US
dc.identifier.issn	0014-4886	en_US
dc.identifier.uri	http://hdl.handle.net/10453/128247
dc.description.abstract	© 2017 Elsevier Inc. Connexin43 (Cx43) hemichannels in spinal cord astrocytes are implicated in the maintenance of neuropathic pain following peripheral nerve injury. Peptide5 is a Cx43 mimetic peptide that blocks hemichannels. In this study, we investigated the effects of spinal delivery of Peptide5 on mechanical pain hypersensitivity in two mouse models of neuropathic pain, peripheral nerve injury and chemotherapy-induced peripheral neuropathy (CIPN). We demonstrated that 10 days following a chronic constriction injury (CCI) of the sciatic nerve, Cx43 expression, co-localised predominantly with astrocytes, was increased in the ipsilateral L3–L5 lumbar spinal cord. An intrathecal injection of Peptide5 into nerve-injured mice, on day 10 when pain was well-established, caused significant improvement in mechanical pain hypersensitivity 8 h after injection. Peptide5 treatment resulted in significantly reduced Cx43, and microglial and astrocyte activity in the dorsal horn of the spinal cord, as compared to control saline-treated CCI mice. Further in vitro investigations on primary astrocyte cultures showed that 1 h pre-treatment with Peptide5 significantly reduced adenosine triphosphate (ATP) release in response to extracellular calcium depletion. Since ATP is a known activator of the NOD-like receptor protein 3 (NLRP3) inflammasome complex, a key mediator of neuroinflammation, we examined the effects of Peptide5 treatment on NLRP3 inflammasome expression. We found that NLRP3, its adaptor apoptosis-associated spec-like protein (ASC) and caspase-1 protein were increased in the ipsilateral spinal cord of CCI mice and reduced to naïve levels following Peptide5 treatment. In the models of oxaliplatin- and paclitaxel-induced peripheral neuropathy, treatment with Peptide5 had no effect on mechanical pain hypersensitivity. Interestingly, in these CIPN models, although spinal Cx43 expression was significantly increased at day 13 following chemotherapy, NLRP3 expression was not altered. These results suggest that the analgesic effect of Peptide5 is specifically achieved by reducing NLRP3 expression. Together, our findings demonstrate that blocking Cx43 hemichannels with Peptide5 after nerve injury attenuates mechanical pain hypersensitivity by specifically targeting the NLRP3 inflammasome in the spinal cord.	en_US
dc.relation.ispartof	Experimental Neurology	en_US
dc.relation.isbasedon	10.1016/j.expneurol.2017.10.016	en_US
dc.subject.classification	Neurology & Neurosurgery	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Mice, Inbred C57BL	en_US
dc.subject.mesh	Mice	en_US
dc.subject.mesh	Neuralgia	en_US
dc.subject.mesh	Hyperalgesia	en_US
dc.subject.mesh	Peptide Fragments	en_US
dc.subject.mesh	Connexin 43	en_US
dc.subject.mesh	Treatment Outcome	en_US
dc.subject.mesh	Injections, Spinal	en_US
dc.subject.mesh	Biomimetic Materials	en_US
dc.subject.mesh	Male	en_US
dc.subject.mesh	Inflammasomes	en_US
dc.subject.mesh	NLR Family, Pyrin Domain-Containing 3 Protein	en_US
dc.title	Attenuation of mechanical pain hypersensitivity by treatment with Peptide5, a connexin-43 mimetic peptide, involves inhibition of NLRP3 inflammasome in nerve-injured mice	en_US
dc.type	Journal Article
utslib.citation.volume	300	en_US
utslib.for	1109 Neurosciences	en_US
utslib.for	1103 Clinical Sciences	en_US
utslib.for	1701 Psychology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
utslib.copyright.status	closed_access
pubs.publication-status	Published	en_US
pubs.volume	300	en_US

Abstract:

© 2017 Elsevier Inc. Connexin43 (Cx43) hemichannels in spinal cord astrocytes are implicated in the maintenance of neuropathic pain following peripheral nerve injury. Peptide5 is a Cx43 mimetic peptide that blocks hemichannels. In this study, we investigated the effects of spinal delivery of Peptide5 on mechanical pain hypersensitivity in two mouse models of neuropathic pain, peripheral nerve injury and chemotherapy-induced peripheral neuropathy (CIPN). We demonstrated that 10 days following a chronic constriction injury (CCI) of the sciatic nerve, Cx43 expression, co-localised predominantly with astrocytes, was increased in the ipsilateral L3–L5 lumbar spinal cord. An intrathecal injection of Peptide5 into nerve-injured mice, on day 10 when pain was well-established, caused significant improvement in mechanical pain hypersensitivity 8 h after injection. Peptide5 treatment resulted in significantly reduced Cx43, and microglial and astrocyte activity in the dorsal horn of the spinal cord, as compared to control saline-treated CCI mice. Further in vitro investigations on primary astrocyte cultures showed that 1 h pre-treatment with Peptide5 significantly reduced adenosine triphosphate (ATP) release in response to extracellular calcium depletion. Since ATP is a known activator of the NOD-like receptor protein 3 (NLRP3) inflammasome complex, a key mediator of neuroinflammation, we examined the effects of Peptide5 treatment on NLRP3 inflammasome expression. We found that NLRP3, its adaptor apoptosis-associated spec-like protein (ASC) and caspase-1 protein were increased in the ipsilateral spinal cord of CCI mice and reduced to naïve levels following Peptide5 treatment. In the models of oxaliplatin- and paclitaxel-induced peripheral neuropathy, treatment with Peptide5 had no effect on mechanical pain hypersensitivity. Interestingly, in these CIPN models, although spinal Cx43 expression was significantly increased at day 13 following chemotherapy, NLRP3 expression was not altered. These results suggest that the analgesic effect of Peptide5 is specifically achieved by reducing NLRP3 expression. Together, our findings demonstrate that blocking Cx43 hemichannels with Peptide5 after nerve injury attenuates mechanical pain hypersensitivity by specifically targeting the NLRP3 inflammasome in the spinal cord.

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http://hdl.handle.net/10453/128247