High Content Positional Biosensor Assay to Screen for Compounds that Prevent or Disrupt Androgen Receptor and...

Area of Science:

• Molecular Biology
• Biochemistry
• Oncology

Background:

• Transcriptional Intermediary Factor 2 (TIF2) is a key coactivator for the Androgen Receptor (AR).
• AR-TIF2 interactions are implicated in the development and progression of castration-resistant prostate cancer (CRPC).
• Targeting AR-TIF2 interactions offers a potential therapeutic strategy for CRPC.

Purpose of the Study:

• To develop and implement a biosensor assay for screening small molecules targeting AR-TIF2 protein-protein interactions (PPIs).
• To identify compounds that can induce, inhibit, or disrupt AR-TIF2 PPIs.
• To provide a cell-based, scalable screening method for potential CRPC therapeutics.

Main Methods:

• Development of an AR-TIF2 protein-protein interaction (PPI) biosensor using fluorescent protein reporters (AR-RFP and TIF2-GFP).
• Utilized high-content imaging and analysis to quantify AR-TIF2 PPIs.
• Employed recombinant adenovirus (rAV) for homogeneous and scalable expression of biosensor components.

Main Results:

• The biosensor assay successfully recapitulates ligand-induced AR translocation and AR-TIF2 complex formation.
• AR-RFP co-localizes with TIF2-GFP in nucleoli upon Dihydrotestosterone (DHT) exposure, indicating successful AR-TIF2 interaction.
• The assay is scalable and suitable for high-throughput screening.

Conclusions:

• The AR-TIF2 PPI biosensor is a promising tool for identifying modulators of AR transcriptional activity.
• This approach may lead to novel therapeutics that overcome resistance mechanisms in CRPC.
• Cell-specific modulation of AR activity offers a targeted strategy against prostate cancer progression.