Journal article
Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer
Cancers, Vol.14(19), p.4883
10/01/2022
DOI: 10.3390/cancers14194883
PMCID: PMC9561963
PMID: 36230806
Abstract
Simple Summary In this study, we generate an innovative endogenous progesterone receptor (PR) reporter gene containing endometrial cancer cell lines and use this tool to visualize PR expression in real-time. We also demonstrated two strategies of using this reporter gene: (1). Drug discovery-screening small molecule inducers for PR expression; (2). Mechanistic investigation-screening potential PR repressors using genome-wide CRISPR knockout library. Expression of progesterone receptor (PR) is a favorable prognostic marker for multiple solid tumors. However, PR expression is reduced or lost in malignant tumors. Thus, monitoring and restoring functional PR expression is important in order to sensitize tumor cells to progesterone therapy in endometrial cancer. We developed stable PR reporter gene containing endometrial cancer cell lines monitoring the endogenous PR expression by inserting mCherry and hygromycin resistant gene at the endogenous PR gene locus by CRISPR/Cas9-mediated genome editing technique. This allows efficient, real-time monitoring of PR expression in its native epigenetic landscape. Reporter gene expression faithfully reflects and amplifies PR expression following treatment with drugs known to induce PR expression. Small molecular PR inducers have been identified from the FDA-approved 1018 drug library and tested for their ability to restore PR expression. Additionally, several candidate PR repressors have been identified by screening the genome-wide CRISPR knockout (GeCKO) library. This novel endogenous PR reporter gene system facilitates the discovery of a new treatment strategy to enhance PR expression and further sensitize progestin therapy in endometrial cancer. These tools provide a systematic, unbiased approach for monitoring target gene expression, allowing for novel drug discovery and mechanistic exploration.
Details
- Title: Subtitle
- Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer
- Creators
- Yiyang Li - Univ Iowa, Dept Obstet & Gynecol, Iowa City, IA 52242 USAWei Zhou - Univ Iowa, Dept Obstet & Gynecol, Iowa City, IA 52242 USAXiangbing Meng - Univ Iowa, Dept Pathol, Iowa City, IA 52242 USASarina D. Murray - Univ Iowa, Dept Pathol, Iowa City, IA 52242 USALong Li - Univ Iowa, Dept Obstet & Gynecol, Iowa City, IA 52242 USAAbby Fronk - Univ Iowa, Dept Obstet & Gynecol, Iowa City, IA 52242 USAVanessa J. Lazaro-Camp - Univ Iowa, Dept Obstet & Gynecol, Iowa City, IA 52242 USAKuo-kuang Wen - High Throughput Screening Facil Univ Iowa UIHTS, Iowa City, IA 52242 USAMeng Wu - High Throughput Screening Facil Univ Iowa UIHTS, Iowa City, IA 52242 USAAdam Dupuy - Univ Iowa, Holden Comprehens Canc Ctr, Iowa City, IA 52242 USAKimberly K. Leslie - Univ Iowa, Dept Obstet & Gynecol, Iowa City, IA 52242 USAShujie Yang - Univ Iowa, Dept Pathol, Iowa City, IA 52242 USA
- Resource Type
- Journal article
- Publication Details
- Cancers, Vol.14(19), p.4883
- DOI
- 10.3390/cancers14194883
- PMID
- 36230806
- PMCID
- PMC9561963
- NLM abbreviation
- Cancers (Basel)
- ISSN
- 2072-6694
- eISSN
- 2072-6694
- Publisher
- Mdpi
- Number of pages
- 20
- Grant note
- Department of Pathology Holden Comprehensive Cancer Center (HCCC) High throughput screening pilot grant R37CA238274; R01CA184101; R01CA99908; R50CA243786 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA HCCC-PACT grant Department of Obstetrics and Gynecology Research Development Fund
- Language
- English
- Date published
- 10/01/2022
- Academic Unit
- Pharmacy; Anatomy and Cell Biology; Stead Family Department of Pediatrics; Pathology; Obstetrics and Gynecology
- Record Identifier
- 9984305894102771
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