Yeast-based evolutionary modeling of androgen receptor mutations and natural selection

Area of Science:

• Molecular Biology
• Genetics
• Biochemistry

Background:

• Prostate cancer (PCa) progression involves genetic mutations in the androgen receptor (AR).
• Understanding AR mutation responsiveness to hormone ligands is crucial but challenging.
• Current annotation lacks comprehensive data on AR mutation-ligand interactions.

Purpose of the Study:

• To develop a rapid yeast reporter system for evaluating androgen receptor (AR) mutations.
• To assess the responsiveness of clinical AR mutations to diverse steroidal ligands.
• To simulate AR gene evolution under selective pressures of steroidal compounds.

Main Methods:

• Utilized a yeast reporter system engineered with an AR-driven histidine synthesis pathway.
• Combined the reporter system with a random DNA mutagenesis library for artificial evolution.
• Screened fifty clinical AR mutations against steroidal ligands like DHT, E2, PROG, and CPA.
• Employed sequential selection to identify multi-point AR mutations.

Main Results:

• Identified 19 AR mutants within 1-2 weeks.
• Validated activation of 11 AR mutants by tested steroidal compounds.
• Demonstrated the system's efficiency in discovering AR mutation evolution directions.
• Successfully simulated AR gene evolution under steroidal ligand pressure.

Conclusions:

• The developed designer yeast system is a portable and efficient module for high-throughput AR-compound screening.
• This system can serve as a valuable clinical reference for prostate cancer (PCa).
• The reporter module can be integrated with other bioassay systems for expanded applications.