Mechanical loading inhibits cartilage inflammatory signalling via an HDAC6 and IFT-dependent mechanism regulating primary cilia elongation
Mechanical loading inhibits cartilage inflammatory signalling via an HDAC6 and IFT-dependent mechanism regulating primary cilia elongation
Objective: physiological mechanical loading reduces inflammatory signalling in numerous cell types including articular chondrocytes however the mechanism responsible remains unclear. This study investigates the role of chondrocyte primary cilia and associated intraflagellar transport (IFT) in the mechanical regulation of interleukin-1β (IL-1β) signalling.
Design: isolated chondrocytes and cartilage explants were subjected to cyclic mechanical loading in the presence and absence of the cytokine IL-1β. Nitric oxide (NO) and prostaglandin E2 (PGE2) release were used to monitor IL-1β signalling whilst Sulphated glycosaminoglycan (sGAG) release provided measurement of cartilage degradation. Measurements were made of HDAC6 activity and tubulin polymerisation and acetylation. Effects on primary cilia were monitored by confocal and super resolution microscopy. Involvement of IFT was analysed using ORPK cells with hypomorphic mutation of IFT88.
Results: mechanical loading suppressed NO and PGE2 release and prevented cartilage degradation. Loading activated HDAC6 and disrupted tubulin acetylation and cilia elongation induced by IL-1β. HDAC6 inhibition with tubacin blocked the anti-inflammatory effects of loading and restored tubulin acetylation and cilia elongation. Hypomorphic mutation of IFT88 reduced IL-1β signalling and abolished the anti-inflammatory effects of loading indicating the mechanism is IFT-dependent. Loading reduced the pool of non-polymerised tubulin which was replicated by taxol which also mimicked the anti-inflammatory effects of mechanical loading and prevented cilia elongation.
Conclusions: this study reveals that mechanical loading suppresses inflammatory signalling, partially dependent on IFT, by activation of HDAC6 and post transcriptional modulation of tubulin.
1064-1074
Fu, S.
f3ae37d7-dad9-49a9-bfd6-8d2ab9b86676
Thompson, C.L.
5f801120-d24b-4a1c-bcd1-8499cce49eb2
Ali, A.
21fb178d-2a61-478a-bedf-66edc27fda62
Wang, W.
8534fc2f-5969-40ab-aff5-96c97efaa922
Chapple, J.P.
662e3573-466a-42c2-b474-039f1cabf406
Mitchison, H.M.
3e88e1ba-29d9-4243-8531-d13e66fa1e01
Beales, P.L.
bf32dc7e-ffd2-4543-8196-a1d340e21707
Wann, A.K.T.
f1b0ea2f-dc8a-4588-a9d8-ae462ed0a993
Knight, M.M.
03701351-a8b9-4b66-8237-fca60c6b5412
13 June 2019
Fu, S.
f3ae37d7-dad9-49a9-bfd6-8d2ab9b86676
Thompson, C.L.
5f801120-d24b-4a1c-bcd1-8499cce49eb2
Ali, A.
21fb178d-2a61-478a-bedf-66edc27fda62
Wang, W.
8534fc2f-5969-40ab-aff5-96c97efaa922
Chapple, J.P.
662e3573-466a-42c2-b474-039f1cabf406
Mitchison, H.M.
3e88e1ba-29d9-4243-8531-d13e66fa1e01
Beales, P.L.
bf32dc7e-ffd2-4543-8196-a1d340e21707
Wann, A.K.T.
f1b0ea2f-dc8a-4588-a9d8-ae462ed0a993
Knight, M.M.
03701351-a8b9-4b66-8237-fca60c6b5412
Fu, S., Thompson, C.L., Ali, A., Wang, W., Chapple, J.P., Mitchison, H.M., Beales, P.L., Wann, A.K.T. and Knight, M.M.
(2019)
Mechanical loading inhibits cartilage inflammatory signalling via an HDAC6 and IFT-dependent mechanism regulating primary cilia elongation.
Osteoarthritis and Cartilage, 27 (7), .
(doi:10.1016/J.JOCA.2019.03.003).
Abstract
Objective: physiological mechanical loading reduces inflammatory signalling in numerous cell types including articular chondrocytes however the mechanism responsible remains unclear. This study investigates the role of chondrocyte primary cilia and associated intraflagellar transport (IFT) in the mechanical regulation of interleukin-1β (IL-1β) signalling.
Design: isolated chondrocytes and cartilage explants were subjected to cyclic mechanical loading in the presence and absence of the cytokine IL-1β. Nitric oxide (NO) and prostaglandin E2 (PGE2) release were used to monitor IL-1β signalling whilst Sulphated glycosaminoglycan (sGAG) release provided measurement of cartilage degradation. Measurements were made of HDAC6 activity and tubulin polymerisation and acetylation. Effects on primary cilia were monitored by confocal and super resolution microscopy. Involvement of IFT was analysed using ORPK cells with hypomorphic mutation of IFT88.
Results: mechanical loading suppressed NO and PGE2 release and prevented cartilage degradation. Loading activated HDAC6 and disrupted tubulin acetylation and cilia elongation induced by IL-1β. HDAC6 inhibition with tubacin blocked the anti-inflammatory effects of loading and restored tubulin acetylation and cilia elongation. Hypomorphic mutation of IFT88 reduced IL-1β signalling and abolished the anti-inflammatory effects of loading indicating the mechanism is IFT-dependent. Loading reduced the pool of non-polymerised tubulin which was replicated by taxol which also mimicked the anti-inflammatory effects of mechanical loading and prevented cilia elongation.
Conclusions: this study reveals that mechanical loading suppresses inflammatory signalling, partially dependent on IFT, by activation of HDAC6 and post transcriptional modulation of tubulin.
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Accepted/In Press date: 16 March 2019
e-pub ahead of print date: 25 March 2019
Published date: 13 June 2019
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Local EPrints ID: 488338
URI: http://eprints.soton.ac.uk/id/eprint/488338
ISSN: 1063-4584
PURE UUID: 60a0faa4-85f7-494b-bbbc-736fb8cf58e8
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Date deposited: 20 Mar 2024 17:59
Last modified: 21 Mar 2024 03:12
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Author:
S. Fu
Author:
C.L. Thompson
Author:
A. Ali
Author:
W. Wang
Author:
J.P. Chapple
Author:
H.M. Mitchison
Author:
P.L. Beales
Author:
A.K.T. Wann
Author:
M.M. Knight
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