Cell-Based Assays to Identify ERR and ERR/PGC Modulators.

Area of Science:

• Endocrinology
• Molecular Biology
• Drug Discovery

Background:

• Estrogen-related receptor alpha (ERRα) is an orphan nuclear receptor involved in endocrine disruption, energy balance, and cancer.
• ERRα function is modulated by cofactors, notably proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α).
• Understanding ERRα's interactions is crucial for developing targeted therapies.

Purpose of the Study:

• To develop a quantitative, high-throughput screening assay for identifying ERRα agonists and antagonists.
• To assess potential toxicity of ERRα modulators.
• To evaluate ERRα activity both independently and in conjunction with PGC-1α.

Main Methods:

• A multiplex luciferase assay was designed and optimized.
• Two distinct stable cell lines were utilized for the assay.
• The protocol allows for simultaneous screening of agonists, antagonists, and toxicity.

Main Results:

• The developed assay provides a quantitative measure of ERRα activity.
• The assay successfully differentiates between agonists and antagonists.
• The system allows for the assessment of ERRα modulation in the presence or absence of PGC-1α.

Conclusions:

• This multiplex luciferase assay is an effective tool for high-throughput screening of ERRα modulators.
• The assay facilitates the identification of potential drug candidates targeting ERRα.
• The protocol enables comprehensive evaluation of ERRα activity and potential toxicity in a single assay.