Characterisation of C101248: A novel selective THIK-1 channel inhibitor for the modulation of microglial NLRP3-inflammasome

Neuroinflammation, specifically the NLRP3 inflammasome cascade, is a common underlying pathological feature of many neurodegenerative diseases. Evidence suggests that NLRP3 activation involves changes in intracellular K+. Nuclear Enriched Transcript Sort Sequencing (NETSseq), which allows for deep s...

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Published in:	Neuropharmacology Vol. 224; p. 109330
Main Authors:	Ossola, Bernardino, Rifat, Ali, Rowland, Anna, Hunter, Helen, Drinkall, Samuel, Bender, Clare, Hamlischer, Mayida, Teall, Martin, Burley, Russell, Barker, Daneil F., Cadwalladr, David, Dickson, Louise, Lawrence, Jason M.K., Harvey, Jenna R.M., Lizio, Marina, Xu, Xiao, Kavanagh, Edel, Cheung, Toni, Sheardown, Steve, Lawrence, Catherine B., Harte, Michael, Brough, David, Madry, Christian, Matthews, Kim, Doyle, Kevin, Page, Keith, Powell, Justin, Brice, Nicola L., Bürli, Roland W., Carlton, Mark B., Dawson, Lee A.
Format:	Journal Article
Language:	English
Published:	England Elsevier Ltd 15-02-2023 Pergamon Press
Subjects:	Alzheimer Disease - metabolism Alzheimer's disease Animals Brain - metabolism Humans Inflammasomes - metabolism KCNK13 Mice Microglia Neuroinflammation Neuroinflammatory Diseases NLR Family, Pyrin Domain-Containing 3 Protein - metabolism NLRP3 inflammasome Potassium Channels, Tandem Pore Domain - antagonists & inhibitors THIK-1 NETSseq AD KCNK13 K2P Neuroinflammation NLRP3 inflammasome IL-1β Microglia DAMPs THIK-1 NLRP3 PAMPs Alzheimer's disease
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Summary:	Neuroinflammation, specifically the NLRP3 inflammasome cascade, is a common underlying pathological feature of many neurodegenerative diseases. Evidence suggests that NLRP3 activation involves changes in intracellular K+. Nuclear Enriched Transcript Sort Sequencing (NETSseq), which allows for deep sequencing of purified cell types from human post-mortem brain tissue, demonstrated a highly specific expression of the tandem pore domain halothane-inhibited K+ channel 1 (THIK-1) in microglia compared to other glial and neuronal cell types in the human brain. NETSseq also showed a significant increase of THIK-1 in microglia isolated from cortical regions of brains with Alzheimer's disease (AD) relative to control donors. Herein, we report the discovery and pharmacological characterisation of C101248, the first selective small-molecule inhibitor of THIK-1. C101248 showed a concentration-dependent inhibition of both mouse and human THIK-1 (IC50: ∼50 nM) and was inactive against K2P family members TREK-1 and TWIK-2, and Kv2.1. Whole-cell patch-clamp recordings of microglia from mouse hippocampal slices showed that C101248 potently blocked both tonic and ATP-evoked THIK-1 K+ currents. Notably, C101248 had no effect on other constitutively active resting conductance in slices from THIK-1-depleted mice. In isolated microglia, C101248 prevented NLRP3-dependent release of IL-1β, an effect not seen in THIK-1-depleted microglia. In conclusion, we demonstrated that inhibiting THIK-1 (a microglia specific gene that is upregulated in brains from donors with AD) using a novel selective modulator attenuates the NLRP3-dependent release of IL-1β from microglia, which suggests that this channel may be a potential therapeutic target for the modulation of neuroinflammation in AD. •THIK-1 mRNA and protein are highly enriched in microglia in human cortical tissue.•THIK-1 mRNA is increased in microglial from AD brains compared to non-AD controls.•Disclosure of C101248, the first selective small-molecule to block THIK-1 currents.•THIK-1 inhibition prevented the NLRP3-dependent release of IL-1β from microglia.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Authors with equal contribution.
ISSN:	0028-3908 1873-7064
DOI:	10.1016/j.neuropharm.2022.109330