A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation
<p>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...
| Main Authors: | , , , , , , , , , |
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| Formato: | Journal article |
| Idioma: | English |
| Publicado em: |
Springer Nature
2021
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| _version_ | 1826309723692990464 |
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| author | Nizami, S Millar, V Arunasalam, K Zarganes-Tzitzikas, T Brough, D Tresadern, G Brennan, PE Davis, JB Ebner, D Di Daniel, E |
| author_facet | Nizami, S Millar, V Arunasalam, K Zarganes-Tzitzikas, T Brough, D Tresadern, G Brennan, PE Davis, JB Ebner, D Di Daniel, E |
| author_sort | Nizami, S |
| collection | OXFORD |
| description | <p>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.</p> |
| first_indexed | 2024-03-07T07:39:58Z |
| format | Journal article |
| id | oxford-uuid:d00136d8-5202-436a-a3d7-569f45f7841b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:39:58Z |
| publishDate | 2021 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | oxford-uuid:d00136d8-5202-436a-a3d7-569f45f7841b2023-04-17T15:52:36ZA phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activationJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:d00136d8-5202-436a-a3d7-569f45f7841bEnglishSymplectic ElementsSpringer Nature2021Nizami, SMillar, VArunasalam, KZarganes-Tzitzikas, TBrough, DTresadern, GBrennan, PEDavis, JBEbner, DDi Daniel, E<p>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.</p> |
| spellingShingle | Nizami, S Millar, V Arunasalam, K Zarganes-Tzitzikas, T Brough, D Tresadern, G Brennan, PE Davis, JB Ebner, D Di Daniel, E A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation |
| title | 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_short | 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 |
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