Development of a High-Throughput Screening Assay for Small-Molecule Inhibitors of Androgen Receptor Splice Variants

Area of Science:

• Oncology
• Molecular Biology
• Drug Discovery

Background:

• Androgen receptor (AR) signaling drives prostate cancer (PCa) progression.
• Current therapies targeting the AR ligand-binding domain (LBD) face resistance due to mutations and splice variants.
• The AR amino-terminal domain (NTD) is crucial for AR function but therapeutically underexplored.

Purpose of the Study:

• To design and develop a functional cell-based assay for identifying small-molecule inhibitors of the AR-NTD.
• To establish a high-throughput screening (HTS) platform for AR-NTD inhibitor discovery.
• To validate initial hits from a targeted library screen in PCa cells.

Main Methods:

• Development of a novel cell-based assay to monitor AR-NTD activity.
• Adaptation of the assay for high-throughput screening.
• Screening of a small, targeted chemical library against the assay.
• Validation of identified hit compounds in prostate cancer cell lines.

Main Results:

• Successful design and implementation of a functional cell-based assay for AR-NTD inhibition.
• Demonstration of the assay's suitability for high-throughput screening.
• Identification and validation of two initial small-molecule inhibitors targeting the AR-NTD from a targeted library screen.

Conclusions:

• The developed cell-based assay is a viable platform for discovering novel AR-NTD inhibitors.
• Targeting the AR-NTD represents a promising new strategy to overcome resistance in advanced prostate cancer.
• Initial validated hits warrant further investigation as potential therapeutics for castration-resistant prostate cancer (CRPC).