Automated high-content live animal drug screening using C. elegans expressing the aggregation prone serpin α1-antitrypsin Z
| Autor: | Tong Ying Shun, David H. Perlmutter, Cliff J. Luke, Paul A. Johnston, Kevin J. Kovatch, Gary A. Silverman, Dale E. King, Olivia S. Long, Joon Hyeok Kwak, John S. Lazo, Sager J. Gosai, Stephen C. Pak |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Drug Evaluation
Preclinical lcsh:Medicine Bioinformatics Biochemistry 0302 clinical medicine Autosomal Recessive Drug Discovery Molecular Cell Biology Fluphenazine Enzyme Inhibitors lcsh:Science Caenorhabditis elegans media_common Cellular Stress Responses 0303 health sciences Multidisciplinary biology Drug discovery Liver Diseases Pimozide Animal Models Phenotype Cirrhosis Autosomal Dominant Cantharidin Models Animal Medicine Research Article Drug Drugs and Devices Drug Research and Development Cell Survival media_common.quotation_subject Transgene Computational biology Gastroenterology and Hepatology Serpin 03 medical and health sciences Model Organisms In vivo Autophagy Animals Humans Viability assay Sodium Azide Biology 030304 developmental biology Clinical Genetics lcsh:R Proteins Acute Phase Proteins biology.organism_classification Luminescent Proteins Microscopy Fluorescence alpha 1-Antitrypsin Dopamine Antagonists lcsh:Q Globular Proteins 030217 neurology & neurosurgery |
| Zdroj: | PLoS ONE, Vol 5, Iss 11, p e15460 (2010) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | The development of preclinical models amenable to live animal bioactive compound screening is an attractive approach to discovering effective pharmacological therapies for disorders caused by misfolded and aggregation-prone proteins. In general, however, live animal drug screening is labor and resource intensive, and has been hampered by the lack of robust assay designs and high throughput work-flows. Based on their small size, tissue transparency and ease of cultivation, the use of C. elegans should obviate many of the technical impediments associated with live animal drug screening. Moreover, their genetic tractability and accomplished record for providing insights into the molecular and cellular basis of human disease, should make C. elegans an ideal model system for in vivo drug discovery campaigns. The goal of this study was to determine whether C. elegans could be adapted to high-throughput and high-content drug screening strategies analogous to those developed for cell-based systems. Using transgenic animals expressing fluorescently-tagged proteins, we first developed a high-quality, high-throughput work-flow utilizing an automated fluorescence microscopy platform with integrated image acquisition and data analysis modules to qualitatively assess different biological processes including, growth, tissue development, cell viability and autophagy. We next adapted this technology to conduct a small molecule screen and identified compounds that altered the intracellular accumulation of the human aggregation prone mutant that causes liver disease in α1-antitrypsin deficiency. This study provides powerful validation for advancement in preclinical drug discovery campaigns by screening live C. elegans modeling α1-antitrypsin deficiency and other complex disease phenotypes on high-content imaging platforms. |
| Databáze: | OpenAIRE |
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