Development of a Phenotypic High-Content Assay to Identify Pharmacoperone Drugs for the Treatment of Primary Hyperoxaluria Type 1 by High-Throughput Screening
| Autor: | Jo Ann Janovick, Sonia Fargue, Louis Scampavia, Christopher J. Danpure, Yih-Tai Chen, David C. Smithson, Timothy P. Spicer, Franck Madoux, P. Michael Conn |
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| Rok vydání: | 2015 |
| Předmět: |
Cell Survival
High-throughput screening Mutant Cell Drug Evaluation Preclinical CHO Cells Biology Mitochondrion Bioinformatics Primary hyperoxaluria Cricetulus Drug Discovery medicine Animals Humans Technology Pharmaceutical Peroxisome medicine.disease Phenotype High-Throughput Screening Assays medicine.anatomical_structure Biochemistry Drug Design Hyperoxaluria Primary Molecular Medicine Biological Assay Function (biology) Molecular Chaperones |
| Zdroj: | ASSAY and Drug Development Technologies. 13:16-24 |
| ISSN: | 1557-8127 1540-658X |
| DOI: | 10.1089/adt.2014.627 |
| Popis: | Primary hyperoxaluria is a severe disease for which the best current therapy is dialysis or organ transplantation. These are risky, inconvenient, and costly procedures. In some patients, pyridoxine treatment can delay the need for these surgical procedures. The underlying cause of particular forms of this disease is the misrouting of a specific enzyme, alanine:glyoxylate aminotransferase (AGT), to the mitochondria instead of the peroxisomes. Pharmacoperones are small molecules that can rescue misfolded proteins and redirect them to their correct location, thereby restoring their function and potentially curing disease. In the present study, we miniaturized a cell-based assay to identify pharmacoperone drugs present in large chemical libraries to selectively correct AGT misrouting. This assay employs AGT-170, a mutant form of AGT that predominantly resides in the mitochondria, which we monitor for its relocation to the peroxisomes through automated image acquisition and analysis. Over the course of a pilot screen of 1,280 test compounds, we achieved an average Z'-factor of 0.72±0.02, demonstrating the suitability of this assay for HTS. |
| Databáze: | OpenAIRE |
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