An unbiased cell morphology-based screen for new, biologically active small molecules.
We have implemented an unbiased cell morphology-based screen to identify small-molecule modulators of cellular processes using the Cytometrix (TM) automated imaging and analysis system. This assay format provides unbiased analysis of morphological effects induced by small molecules by capturing phen...
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| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2005-05-01
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| Series: | PLoS Biology |
| Online Access: | https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.0030128&type=printable |
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| _version_ | 1823856912309944320 |
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| author | Masahiro Tanaka Raynard Bateman Daniel Rauh Eugeni Vaisberg Shyam Ramachandani Chao Zhang Kirk C Hansen Alma L Burlingame Jay K Trautman Kevan M Shokat Cynthia L Adams |
| author_facet | Masahiro Tanaka Raynard Bateman Daniel Rauh Eugeni Vaisberg Shyam Ramachandani Chao Zhang Kirk C Hansen Alma L Burlingame Jay K Trautman Kevan M Shokat Cynthia L Adams |
| author_sort | Masahiro Tanaka |
| collection | DOAJ |
| description | We have implemented an unbiased cell morphology-based screen to identify small-molecule modulators of cellular processes using the Cytometrix (TM) automated imaging and analysis system. This assay format provides unbiased analysis of morphological effects induced by small molecules by capturing phenotypic readouts of most known classes of pharmacological agents and has the potential to read out pathways for which little is known. Four human-cancer cell lines and one noncancerous primary cell type were treated with 107 small molecules comprising four different protein kinase-inhibitor scaffolds. Cellular phenotypes induced by each compound were quantified by multivariate statistical analysis of the morphology, staining intensity, and spatial attributes of the cellular nuclei, microtubules, and Golgi compartments. Principal component analysis was used to identify inhibitors of cellular components not targeted by known protein kinase inhibitors. Here we focus on a hydroxyl-substituted analog (hydroxy-PP) of the known Src-family kinase inhibitor PP2 because it induced cell-specific morphological features distinct from all known kinase inhibitors in the collection. We used affinity purification to identify a target of hydroxy-PP, carbonyl reductase 1 (CBR1), a short-chain dehydrogenase-reductase. We solved the X-ray crystal structure of the CBR1/hydroxy-PP complex to 1.24 A resolution. Structure-based design of more potent and selective CBR1 inhibitors provided probes for analyzing the biological function of CBR1 in A549 cells. These studies revealed a previously unknown function for CBR1 in serum-withdrawal-induced apoptosis. Further studies indicate CBR1 inhibitors may enhance the effectiveness of anticancer anthracyclines. Morphology-based screening of diverse cancer cell types has provided a method for discovering potent new small-molecule probes for cell biological studies and anticancer drug candidates. |
| format | Article |
| id | doaj-art-793028f6ead843649a23bfd14fe7747d |
| institution | Kabale University |
| issn | 1544-9173 1545-7885 |
| language | English |
| publishDate | 2005-05-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj-art-793028f6ead843649a23bfd14fe7747d2025-02-12T05:30:22ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852005-05-0135e12810.1371/journal.pbio.0030128An unbiased cell morphology-based screen for new, biologically active small molecules.Masahiro TanakaRaynard BatemanDaniel RauhEugeni VaisbergShyam RamachandaniChao ZhangKirk C HansenAlma L BurlingameJay K TrautmanKevan M ShokatCynthia L AdamsWe have implemented an unbiased cell morphology-based screen to identify small-molecule modulators of cellular processes using the Cytometrix (TM) automated imaging and analysis system. This assay format provides unbiased analysis of morphological effects induced by small molecules by capturing phenotypic readouts of most known classes of pharmacological agents and has the potential to read out pathways for which little is known. Four human-cancer cell lines and one noncancerous primary cell type were treated with 107 small molecules comprising four different protein kinase-inhibitor scaffolds. Cellular phenotypes induced by each compound were quantified by multivariate statistical analysis of the morphology, staining intensity, and spatial attributes of the cellular nuclei, microtubules, and Golgi compartments. Principal component analysis was used to identify inhibitors of cellular components not targeted by known protein kinase inhibitors. Here we focus on a hydroxyl-substituted analog (hydroxy-PP) of the known Src-family kinase inhibitor PP2 because it induced cell-specific morphological features distinct from all known kinase inhibitors in the collection. We used affinity purification to identify a target of hydroxy-PP, carbonyl reductase 1 (CBR1), a short-chain dehydrogenase-reductase. We solved the X-ray crystal structure of the CBR1/hydroxy-PP complex to 1.24 A resolution. Structure-based design of more potent and selective CBR1 inhibitors provided probes for analyzing the biological function of CBR1 in A549 cells. These studies revealed a previously unknown function for CBR1 in serum-withdrawal-induced apoptosis. Further studies indicate CBR1 inhibitors may enhance the effectiveness of anticancer anthracyclines. Morphology-based screening of diverse cancer cell types has provided a method for discovering potent new small-molecule probes for cell biological studies and anticancer drug candidates.https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.0030128&type=printable |
| spellingShingle | Masahiro Tanaka Raynard Bateman Daniel Rauh Eugeni Vaisberg Shyam Ramachandani Chao Zhang Kirk C Hansen Alma L Burlingame Jay K Trautman Kevan M Shokat Cynthia L Adams An unbiased cell morphology-based screen for new, biologically active small molecules. PLoS Biology |
| title | An unbiased cell morphology-based screen for new, biologically active small molecules. |
| title_full | An unbiased cell morphology-based screen for new, biologically active small molecules. |
| title_fullStr | An unbiased cell morphology-based screen for new, biologically active small molecules. |
| title_full_unstemmed | An unbiased cell morphology-based screen for new, biologically active small molecules. |
| title_short | An unbiased cell morphology-based screen for new, biologically active small molecules. |
| title_sort | unbiased cell morphology based screen for new biologically active small molecules |
| url | https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.0030128&type=printable |
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