High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model.
To identify Huntington's Disease therapeutics, we conducted high-content small molecule and RNAi suppressor screens using a Drosophila primary neural culture Huntingtin model. Drosophila primary neurons offer a sensitive readout for neurotoxicty, as their neurites develop dysmorphic features in...
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| Format: | Article |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0023841 |
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| author | Joost Schulte Katharine J Sepp Chaohong Wu Pengyu Hong J Troy Littleton |
| author_facet | Joost Schulte Katharine J Sepp Chaohong Wu Pengyu Hong J Troy Littleton |
| author_sort | Joost Schulte |
| collection | DOAJ |
| description | To identify Huntington's Disease therapeutics, we conducted high-content small molecule and RNAi suppressor screens using a Drosophila primary neural culture Huntingtin model. Drosophila primary neurons offer a sensitive readout for neurotoxicty, as their neurites develop dysmorphic features in the presence of mutant polyglutamine-expanded Huntingtin compared to nonpathogenic Huntingtin. By tracking the subcellular distribution of mRFP-tagged pathogenic Huntingtin and assaying neurite branch morphology via live-imaging, we identified suppressors that could reduce Huntingtin aggregation and/or prevent the formation of dystrophic neurites. The custom algorithms we used to quantify neurite morphologies in complex cultures provide a useful tool for future high-content screening approaches focused on neurodegenerative disease models. Compounds previously found to be effective aggregation inhibitors in mammalian systems were also effective in Drosophila primary cultures, suggesting translational capacity between these models. However, we did not observe a direct correlation between the ability of a compound or gene knockdown to suppress aggregate formation and its ability to rescue dysmorphic neurites. Only a subset of aggregation inhibitors could revert dysmorphic cellular profiles. We identified lkb1, an upstream kinase in the mTOR/Insulin pathway, and four novel drugs, Camptothecin, OH-Camptothecin, 18β-Glycyrrhetinic acid, and Carbenoxolone, that were strong suppressors of mutant Huntingtin-induced neurotoxicity. Huntingtin neurotoxicity suppressors identified through our screen also restored viability in an in vivo Drosophila Huntington's Disease model, making them attractive candidates for further therapeutic evaluation. |
| format | Article |
| id | doaj-art-7c8eb75c7a5c47d382a7bdd6870beeac |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj-art-7c8eb75c7a5c47d382a7bdd6870beeac2025-08-23T05:32:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0168e2384110.1371/journal.pone.0023841High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model.Joost SchulteKatharine J SeppChaohong WuPengyu HongJ Troy LittletonTo identify Huntington's Disease therapeutics, we conducted high-content small molecule and RNAi suppressor screens using a Drosophila primary neural culture Huntingtin model. Drosophila primary neurons offer a sensitive readout for neurotoxicty, as their neurites develop dysmorphic features in the presence of mutant polyglutamine-expanded Huntingtin compared to nonpathogenic Huntingtin. By tracking the subcellular distribution of mRFP-tagged pathogenic Huntingtin and assaying neurite branch morphology via live-imaging, we identified suppressors that could reduce Huntingtin aggregation and/or prevent the formation of dystrophic neurites. The custom algorithms we used to quantify neurite morphologies in complex cultures provide a useful tool for future high-content screening approaches focused on neurodegenerative disease models. Compounds previously found to be effective aggregation inhibitors in mammalian systems were also effective in Drosophila primary cultures, suggesting translational capacity between these models. However, we did not observe a direct correlation between the ability of a compound or gene knockdown to suppress aggregate formation and its ability to rescue dysmorphic neurites. Only a subset of aggregation inhibitors could revert dysmorphic cellular profiles. We identified lkb1, an upstream kinase in the mTOR/Insulin pathway, and four novel drugs, Camptothecin, OH-Camptothecin, 18β-Glycyrrhetinic acid, and Carbenoxolone, that were strong suppressors of mutant Huntingtin-induced neurotoxicity. Huntingtin neurotoxicity suppressors identified through our screen also restored viability in an in vivo Drosophila Huntington's Disease model, making them attractive candidates for further therapeutic evaluation.https://doi.org/10.1371/journal.pone.0023841 |
| spellingShingle | Joost Schulte Katharine J Sepp Chaohong Wu Pengyu Hong J Troy Littleton High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model. PLoS ONE |
| title | High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model. |
| title_full | High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model. |
| title_fullStr | High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model. |
| title_full_unstemmed | High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model. |
| title_short | High-content chemical and RNAi screens for suppressors of neurotoxicity in a Huntington's disease model. |
| title_sort | high content chemical and rnai screens for suppressors of neurotoxicity in a huntington s disease model |
| url | https://doi.org/10.1371/journal.pone.0023841 |
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