Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer
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 endome...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-10-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/19/4883 |
| _version_ | 1797480126009573376 |
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| author | Yiyang Li Wei Zhou Xiangbing Meng Sarina D. Murray Long Li Abby Fronk Vanessa J. Lazaro-Camp Kuo-kuang Wen Meng Wu Adam Dupuy Kimberly K. Leslie Shujie Yang |
| author_facet | Yiyang Li Wei Zhou Xiangbing Meng Sarina D. Murray Long Li Abby Fronk Vanessa J. Lazaro-Camp Kuo-kuang Wen Meng Wu Adam Dupuy Kimberly K. Leslie Shujie Yang |
| author_sort | Yiyang Li |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T21:55:20Z |
| format | Article |
| id | doaj.art-0490516fc24c47a4a559db8b8dbd0d78 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-09T21:55:20Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-0490516fc24c47a4a559db8b8dbd0d782023-11-23T19:58:12ZengMDPI AGCancers2072-66942022-10-011419488310.3390/cancers14194883Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial CancerYiyang Li0Wei Zhou1Xiangbing Meng2Sarina D. Murray3Long Li4Abby Fronk5Vanessa J. Lazaro-Camp6Kuo-kuang Wen7Meng Wu8Adam Dupuy9Kimberly K. Leslie10Shujie Yang11Department of Obstetrics and Gynecology, The University of Iowa, Iowa City, IA 52242, USADepartment of Obstetrics and Gynecology, The University of Iowa, Iowa City, IA 52242, USADepartment of Pathology, The University of Iowa, Iowa City, IA 52242, USADepartment of Pathology, The University of Iowa, Iowa City, IA 52242, USADepartment of Obstetrics and Gynecology, The University of Iowa, Iowa City, IA 52242, USADepartment of Obstetrics and Gynecology, The University of Iowa, Iowa City, IA 52242, USADepartment of Obstetrics and Gynecology, The University of Iowa, Iowa City, IA 52242, USAHigh Throughput Screening Facility at University of Iowa (UIHTS), Iowa City, IA 52242, USAHigh Throughput Screening Facility at University of Iowa (UIHTS), Iowa City, IA 52242, USAHolden Comprehensive Cancer Center, University of Iowa, Iowa City, IA 52242, USADepartment of Obstetrics and Gynecology, The University of Iowa, Iowa City, IA 52242, USADepartment of Pathology, The University of Iowa, Iowa City, IA 52242, USAExpression 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.https://www.mdpi.com/2072-6694/14/19/4883progesterone receptormCherryreporter geneendometrial cancerhigh throughput screeningGeCKO library |
| spellingShingle | Yiyang Li Wei Zhou Xiangbing Meng Sarina D. Murray Long Li Abby Fronk Vanessa J. Lazaro-Camp Kuo-kuang Wen Meng Wu Adam Dupuy Kimberly K. Leslie Shujie Yang Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer Cancers progesterone receptor mCherry reporter gene endometrial cancer high throughput screening GeCKO library |
| title | Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer |
| title_full | Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer |
| title_fullStr | Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer |
| title_full_unstemmed | Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer |
| title_short | Utilizing an Endogenous Progesterone Receptor Reporter Gene for Drug Screening and Mechanistic Study in Endometrial Cancer |
| title_sort | utilizing an endogenous progesterone receptor reporter gene for drug screening and mechanistic study in endometrial cancer |
| topic | progesterone receptor mCherry reporter gene endometrial cancer high throughput screening GeCKO library |
| url | https://www.mdpi.com/2072-6694/14/19/4883 |
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