Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial...

Area of Science:

• Oncology
• Molecular Biology
• Genetics

Background:

• Progesterone receptor (PR) expression is a positive prognostic indicator in various solid tumors, but it is often diminished in malignant cancers.
• Restoring functional PR expression is crucial for enhancing the efficacy of progesterone therapy in endometrial cancer.

Purpose of the Study:

• To develop a novel system for real-time monitoring of endogenous progesterone receptor (PR) expression in endometrial cancer.
• To identify small molecules that can induce PR expression and potential PR repressors for therapeutic applications.

Main Methods:

• Utilized CRISPR/Cas9 genome editing to create stable endometrial cancer cell lines with a PR reporter gene (mCherry and hygromycin resistance) at the endogenous PR locus.
• Screened the FDA-approved 1018 drug library to identify PR inducers and employed a genome-wide CRISPR knockout (GeCKO) library to find PR repressors.

Main Results:

• The developed reporter gene system accurately reflects and amplifies endogenous PR expression in response to therapeutic agents.
• Identified several small molecules capable of inducing PR expression and potential repressors of PR.
• Demonstrated the utility of the reporter system for drug discovery and mechanistic studies.

Conclusions:

• The novel endogenous PR reporter gene system offers a powerful tool for monitoring PR expression and discovering new therapeutic strategies in endometrial cancer.
• This approach facilitates the identification of treatments to enhance PR levels, thereby sensitizing cancer cells to progestin therapy.