The "TSH Receptor Glo Assay" - A High-Throughput Detection System for Thyroid Stimulation

Area of Science:

• Endocrinology
• Molecular Pharmacology

Background:

• Development of a sensitive transcription-based luciferase high-throughput screening (HTS) system, the TSHR-Glo Assay, for identifying TSH receptor modulators.
• Utilizes double-transfected Chinese hamster ovary cells expressing human TSHR and a CRE-luciferase reporter.

Purpose of the Study:

• To discover novel small molecules that act as agonists for the TSH receptor.
• To establish a robust HTS assay for TSH receptor activity and antibody detection.

Main Methods:

• Screening of 48,224 diverse compounds using the TSHR-Glo assay at a fixed dose.
• Validation of assay performance using the Z-factor, achieving a score of 0.895.
• Secondary screening of active compounds against the parent cell line to eliminate false positives.

Main Results:

• The TSHR-Glo assay demonstrated high responsiveness to TSH (sensitivity 10(-10)M) and detected thyroid-stimulating antibodies.
• Identified 62 potential active molecules from the initial screen of 48,224 compounds.
• Characterized several novel small molecule TSH receptor agonists.

Conclusions:

• The TSHR-Glo assay is an effective one-step, cell-based HTS method for discovering TSH receptor agonists.
• The assay successfully identified specific TSH receptor agonists and can detect stimulating antibodies, relevant for Graves' disease research.