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Development of a High-Throughput Screening Assay for Small-Molecule Inhibitors of Androgen Receptor Splice Variants
Amy E Monaghan1, Alison Porter2, Irene Hunter1
1Institute of Medical Sciences, School of Medicine, Medical Sciences and Nutrition, University of Aberdeen, Aberdeen, United Kingdom.
Abstract:
The role of the androgen receptor (AR) in the progression of prostate cancer (PCa) is well established and competitive inhibition of AR ligand binding domain (LBD) has been the mainstay of antiandrogen therapies for advanced and metastatic disease. However, the efficacy of such drugs is often limited by the emergence of resistance, mediated through point mutations and receptor splice variants lacking the AR-LBD. As a result, the prognosis for patients with malignant, castrate-resistant disease remains poor. The amino terminal domain (NTD) of the AR has been shown to be critical for AR function. Its modular activation function (AF-1) is important for both gene regulation and participation in protein-protein interactions. However, due to the intrinsically disordered structure of the domain, its potential as a candidate for therapeutic intervention has been generally overlooked. In this article, we describe the design and development of a functional cell-based assay aimed at identifying small-molecule inhibitors of the AR-NTD. We demonstrate the suitability of the assay for high-throughput screening platforms and validate two initial hits emerging from a small, targeted, library screen in PCa cells.
Insights
Researchers developed a new assay to find drugs targeting the androgen receptor's amino terminal domain (AR-NTD). This approach aims to overcome resistance to current prostate cancer (PCa) therapies.
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).
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