Curran, Niamh and Griffin, Bryan D. and O'Toole, Daniel and Brady, Kevin J. and Fitzgerald, Stephen and Moynagh, Paul N. (2005) The Synthetic Cannabinoid R(+)WIN 55,212-2 Inhibits the Interleukin-1 Signaling Pathway in Human Astrocytes in a Cannabinoid Receptor-independent Manner. Journal of Biological Chemistry, 28 (43). pp. 35797-35806. ISSN 0021-9258

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Abstract

R(+)WIN 55,212-2 is a synthetic cannabinoid that controls disease progression in models of multiple sclerosis. This is associated with its ability to reduce migration of leukocytes into the central nervous system. Because leukocyte migration is dependent on induction of adhesion molecules and chemokines by pro-inflammatory cytokines, we examined the effects of R(+)WIN 55,212-2 on their expression. Using 1321N1 astrocytoma and A-172 glioblastoma as cell models we show that R(+)WIN 55,212-2, but not its inactive chiral form S(–)WIN 55,212-2, strongly inhibits the interleukin-1 (IL-1) induction of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) and the chemokine IL-8. This inhibition is not mediated via the CB1 or CB2 cannabinoid receptors, because their selective antagonists and pertussis toxin failed to affect the inhibitory effects of R(+)WIN 55,212-2. Furthermore reverse transcription-PCR analysis did not detect the expression of either receptor in 1321N1 cells. R(+)WIN 55,212-2 was shown to inhibit adhesion molecule and chemokine expression at the level of transcription, because it strongly inhibited the IL-1 induction of ICAM-1, VCAM-1, and IL-8 mRNAs and blocked the IL-1 activation of their promoters. The NFκB pathway was then assessed as a lead target for R(+)WIN 55,212-2. NFκB was measured by expression of a transfected NFκB-regulated reporter gene. Using this assay, R(+)WIN 55,212-2 strongly inhibited IL-1 activation of NFκB. Furthermore R(+)WIN 55,212-2 inhibited the ability of overexpressed Myd88, Tak-1, and IKK-2 to induce the reporter gene suggesting that R(+)WIN 55,212-2 acts at or downstream of IKK-2 in the IL-1 pathway. However R(+)WIN 55,212-2 failed to inhibit IL-1-induced degradation of IκBα, excluding IKK-2 as a direct target. In addition electrophoretic mobility shift and chromatin immunoprecipitation assays showed that R(+)WIN 55,212-2 does not regulate the IL-1-induced nuclear translocation of NFκB or the ability of the latter to bind to promoters regulating expression of ICAM-1 and IL-8. These data suggest that R(+)WIN 55,212-2 blocks IL-1 signaling by inhibiting the transactivation potential of NFκB.

Item Type:	Article
Additional Information:	This research was originally published in the Journal of Biological Chemistry. Curran, N.M., Griffin, B.D., O’Toole, D., Brady, K. J., Fitzgerald, S.N. and Moynagh, P.N. (2005) 'The synthetic cannabinoid R(+)WIN55,212 inhibits the interleukin-1 signaling pathway in human astrocytes in a cannabinoid-receptor independent manner'. Journal of Biological Chemistry, 280 :35797-35806
Keywords:	Synthetic Cannabinoid R(+)WIN 55,212-2; Interleukin-1; Signaling Pathway; Human Astrocytes; Cannabinoid Receptor-independent Manner;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	7206
Identification Number:	https://doi.org/10.1074/jbc.M507959200
Depositing User:	Professor Paul Moynagh
Date Deposited:	20 Jul 2016 16:02
Journal or Publication Title:	Journal of Biological Chemistry
Publisher:	American Society for Biochemistry and Molecular Biology
Refereed:	Yes
Funders:	Enterprise Ireland (EI), Health Research Board (HRB), Higher Education Authority (HEA), European Commission
URI:	http://www.jbc.org
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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