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A cell-based high-throughput screening assay for Farnesoid X receptor agonists
Zhi-Hui Zheng1, Guo-Ping Lv, Shu-Yi Si
1Insitute of Medicinal Biotechnology, Peking Union Medical College / Chinese Academy of Medical Sciences, Beijing 100050, China. z_zhihui2003@yahoo.com.cn
Biomedical and Environmental Sciences : BES
|March 20, 2008
Summary
A new high-throughput screening assay was developed to identify Farnesoid X receptor (FXR) agonists. This cell-based model efficiently screens compounds for potential dyslipidemia drug discovery.
Area of Science:
- Pharmacology
- Molecular Biology
- Drug Discovery
Background:
- The Farnesoid X receptor (FXR) plays a crucial role in regulating lipid and glucose metabolism.
- Identifying novel FXR agonists is a key strategy for developing treatments for dyslipidemia.
Purpose of the Study:
- To establish a robust high-throughput screening (HTS) assay for FXR agonists.
- To utilize a mammalian one-hybrid system for identifying potential dyslipidemia drug leads from chemical libraries.
Main Methods:
- Constructed a chimera expression plasmid (GAL4-FXR LBD) by fusing human FXR LBD cDNA to yeast GAL4 DBD.
- Developed a reporter plasmid (pG5-SV40 Luc) containing five GAL4 DNA binding sites upstream of the SV40 promoter.
- Optimized the assay through transient co-transfection of the reporter and expression plasmids.
Main Results:
- The assay demonstrated dose-dependent induction of luciferase gene expression by CDCA, a natural FXR agonist.
- Achieved a signal-to-noise ratio of 10 and a Z' factor of 0.65, indicating assay robustness.
- Validated the assay's sensitivity and stability for screening.
Conclusions:
- A stable and sensitive cell-based HTS model for FXR agonists has been successfully developed.
- This model is suitable for screening both synthetic and natural compound libraries for FXR agonists.
- The assay facilitates the identification of novel lead compounds for dyslipidemia drug development.
