A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation

Abstract Inhibition of the NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome has recently emerged as a promising therapeutic target for several inflammatory diseases. After priming and activation by inflammation triggers, NLRP3 forms a complex with apoptosis-associated speck-like...

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Autores principales:	Sohaib Nizami, Val Millar, Kanisa Arunasalam, Tryfon Zarganes-Tzitzikas, David Brough, Gary Tresadern, Paul E. Brennan, John B. Davis, Daniel Ebner, Elena Di Daniel
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1fa617be6c444588bad302d2d3607a84
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spelling	oai:doaj.org-article:1fa617be6c444588bad302d2d3607a842021-12-02T16:24:52ZA phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation10.1038/s41598-021-94850-w2045-2322https://doaj.org/article/1fa617be6c444588bad302d2d3607a842021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94850-whttps://doaj.org/toc/2045-2322Abstract Inhibition of the NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome has recently emerged as a promising therapeutic target for several inflammatory diseases. After priming and activation by inflammation triggers, NLRP3 forms a complex with apoptosis-associated speck-like protein containing a CARD domain (ASC) followed by formation of the active inflammasome. Identification of inhibitors of NLRP3 activation requires a well-validated primary high-throughput assay followed by the deployment of a screening cascade of assays enabling studies of structure–activity relationship, compound selectivity and efficacy in disease models. We optimized a NLRP3-dependent fluorescent tagged ASC speck formation assay in murine immortalized bone marrow-derived macrophages and utilized it to screen a compound library of 81,000 small molecules. Our high-content screening assay yielded robust assay metrics and identified a number of inhibitors of NLRP3-dependent ASC speck formation, including compounds targeting HSP90, JAK and IKK-β. Additional assays to investigate inflammasome priming or activation, NLRP3 downstream effectors such as caspase-1, IL-1β and pyroptosis form the basis of a screening cascade to identify NLRP3 inflammasome inhibitors in drug discovery programs.Sohaib NizamiVal MillarKanisa ArunasalamTryfon Zarganes-TzitzikasDavid BroughGary TresadernPaul E. BrennanJohn B. DavisDaniel EbnerElena Di DanielNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sohaib Nizami Val Millar Kanisa Arunasalam Tryfon Zarganes-Tzitzikas David Brough Gary Tresadern Paul E. Brennan John B. Davis Daniel Ebner Elena Di Daniel A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
description	Abstract Inhibition of the NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome has recently emerged as a promising therapeutic target for several inflammatory diseases. After priming and activation by inflammation triggers, NLRP3 forms a complex with apoptosis-associated speck-like protein containing a CARD domain (ASC) followed by formation of the active inflammasome. Identification of inhibitors of NLRP3 activation requires a well-validated primary high-throughput assay followed by the deployment of a screening cascade of assays enabling studies of structure–activity relationship, compound selectivity and efficacy in disease models. We optimized a NLRP3-dependent fluorescent tagged ASC speck formation assay in murine immortalized bone marrow-derived macrophages and utilized it to screen a compound library of 81,000 small molecules. Our high-content screening assay yielded robust assay metrics and identified a number of inhibitors of NLRP3-dependent ASC speck formation, including compounds targeting HSP90, JAK and IKK-β. Additional assays to investigate inflammasome priming or activation, NLRP3 downstream effectors such as caspase-1, IL-1β and pyroptosis form the basis of a screening cascade to identify NLRP3 inflammasome inhibitors in drug discovery programs.
format	article
author	Sohaib Nizami Val Millar Kanisa Arunasalam Tryfon Zarganes-Tzitzikas David Brough Gary Tresadern Paul E. Brennan John B. Davis Daniel Ebner Elena Di Daniel
author_facet	Sohaib Nizami Val Millar Kanisa Arunasalam Tryfon Zarganes-Tzitzikas David Brough Gary Tresadern Paul E. Brennan John B. Davis Daniel Ebner Elena Di Daniel
author_sort	Sohaib Nizami
title	A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
title_short	A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
title_full	A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
title_fullStr	A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
title_full_unstemmed	A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
title_sort	phenotypic high-content, high-throughput screen identifies inhibitors of nlrp3 inflammasome activation
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1fa617be6c444588bad302d2d3607a84
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A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation

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