Integrating High-Content Analysis into a Multiplexed Screening Approach to Identify and Characterize GPCR Agonists
| Autor: | Jonathan O’Connell, Martyn Banks, Jenny Xie, John Watson, Neil T. Burford, Mary Ellen Cvijic, Alaric J. Dyckman, Andrew Alt, Mengjie Chen, Yingjie Zhu, Andrea D. Weston, Sukhanya Jayachandra, Litao Zhang, Michele Agler, Ding Ren Shen, Melissa Yarde |
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| Rok vydání: | 2013 |
| Předmět: |
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Allosteric regulation Green Fluorescent Proteins Computational biology CHO Cells Biology Bioinformatics Biochemistry Analytical Chemistry Receptors G-Protein-Coupled Cricetulus Cell Line Tumor Cricetinae Drug Discovery Functional selectivity Cyclic AMP Image Processing Computer-Assisted Animals Humans Internalization beta-Arrestins G protein-coupled receptor media_common Drug discovery Reproducibility of Results Cell based assays Rats Protein Transport Receptors Lysosphingolipid Microscopy Fluorescence High-content screening Molecular Medicine Biological Assay Allosteric Site Biotechnology Protein Binding |
| Zdroj: | Journal of biomolecular screening. 19(7) |
| ISSN: | 1552-454X |
| Popis: | G protein-coupled receptors (GPCRs) are one of the most popular and proven target classes for therapeutic intervention. The increased appreciation for allosteric modulation, receptor oligomerization, and biased agonism has led to the development of new assay platforms that seek to capitalize on these aspects of GPCR biology. High-content screening is particularly well suited for GPCR drug discovery given the ability to image and quantify changes in multiple cellular parameters, to resolve subcellular structures, and to monitor events within a physiologically relevant environment. Focusing on the sphingosine-1-phosphate (S1P1) receptor, we evaluated the utility of high-content approaches in hit identification efforts by developing and applying assays to monitor β-arrestin translocation, GPCR internalization, and GPCR recycling kinetics. Using these approaches in combination with more traditional GPCR screening assays, we identified compounds whose unique pharmacological profiles would have gone unnoticed if using a single platform. In addition, we identified a compound that induces an atypical pattern of β-arrestin translocation and GPCR recycling kinetics. Our results highlight the value of high-content imaging in GPCR drug discovery efforts and emphasize the value of a multiassay approach to study pharmacological properties of compounds of interest. |
| Databáze: | OpenAIRE |
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