TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells
Mutations in components of the Wnt/β-catenin signaling pathway drive colorectal cancer (CRC), in part, by deregulating expression of genes controlled by the T-cell factor (TCF) family of transcription factors. TCFs contain a conserved DNA binding domain that mediates association with TCF binding ele...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-02-01
|
| Series: | Genes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4425/14/2/481 |
| _version_ | 1797620813078200320 |
|---|---|
| author | Carli M. King Olivia M. Marx Wei Ding Walter A. Koltun Gregory S. Yochum |
| author_facet | Carli M. King Olivia M. Marx Wei Ding Walter A. Koltun Gregory S. Yochum |
| author_sort | Carli M. King |
| collection | DOAJ |
| description | Mutations in components of the Wnt/β-catenin signaling pathway drive colorectal cancer (CRC), in part, by deregulating expression of genes controlled by the T-cell factor (TCF) family of transcription factors. TCFs contain a conserved DNA binding domain that mediates association with TCF binding elements (TBEs) within Wnt-responsive DNA elements (WREs). Intestinal stem cell marker, leucine-rich-repeat containing G-protein-coupled receptor 5 (LGR5), is a Wnt target gene that has been implicated in CRC stem cell plasticity. However, the WREs at the <i>LGR5</i> gene locus and how TCF factors directly regulate <i>LGR5</i> gene expression in CRC have not been fully defined. Here, we report that TCF family member, TCF7L1, plays a significant role in regulating <i>LGR5</i> expression in CRC cells. We demonstrate that TCF7L1 binds to a novel promoter-proximal WRE through association with a consensus TBE at the <i>LGR5</i> locus to repress <i>LGR5</i> expression. Using CRISPR activation and interference (CRISPRa/i) technologies to direct epigenetic modulation, we demonstrate that this WRE is a critical regulator of <i>LGR5</i> expression and spheroid formation capacity of CRC cells. Furthermore, we found that restoring <i>LGR5</i> expression rescues the TCF7L1-mediated reduction in spheroid formation efficiency. These results demonstrate a role for TCF7L1 in repressing <i>LGR5</i> gene expression to govern the spheroid formation potential of CRC cells. |
| first_indexed | 2024-03-11T08:46:58Z |
| format | Article |
| id | doaj.art-737c8526c44f48fca5e3fb4e80cfb7ac |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-11T08:46:58Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-737c8526c44f48fca5e3fb4e80cfb7ac2023-11-16T20:43:34ZengMDPI AGGenes2073-44252023-02-0114248110.3390/genes14020481TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer CellsCarli M. King0Olivia M. Marx1Wei Ding2Walter A. Koltun3Gregory S. Yochum4Department of Biochemistry & Molecular Biology, College of Medicine, The Pennsylvania State University, Hershey, PA 17036, USADepartment of Biochemistry & Molecular Biology, College of Medicine, The Pennsylvania State University, Hershey, PA 17036, USADepartment of Surgery, Division of Colon & Rectal Surgery, Milton S. Hershey Medical Center, The Pennsylvania State University, Hershey, PA 17036, USADepartment of Surgery, Division of Colon & Rectal Surgery, Milton S. Hershey Medical Center, The Pennsylvania State University, Hershey, PA 17036, USADepartment of Biochemistry & Molecular Biology, College of Medicine, The Pennsylvania State University, Hershey, PA 17036, USAMutations in components of the Wnt/β-catenin signaling pathway drive colorectal cancer (CRC), in part, by deregulating expression of genes controlled by the T-cell factor (TCF) family of transcription factors. TCFs contain a conserved DNA binding domain that mediates association with TCF binding elements (TBEs) within Wnt-responsive DNA elements (WREs). Intestinal stem cell marker, leucine-rich-repeat containing G-protein-coupled receptor 5 (LGR5), is a Wnt target gene that has been implicated in CRC stem cell plasticity. However, the WREs at the <i>LGR5</i> gene locus and how TCF factors directly regulate <i>LGR5</i> gene expression in CRC have not been fully defined. Here, we report that TCF family member, TCF7L1, plays a significant role in regulating <i>LGR5</i> expression in CRC cells. We demonstrate that TCF7L1 binds to a novel promoter-proximal WRE through association with a consensus TBE at the <i>LGR5</i> locus to repress <i>LGR5</i> expression. Using CRISPR activation and interference (CRISPRa/i) technologies to direct epigenetic modulation, we demonstrate that this WRE is a critical regulator of <i>LGR5</i> expression and spheroid formation capacity of CRC cells. Furthermore, we found that restoring <i>LGR5</i> expression rescues the TCF7L1-mediated reduction in spheroid formation efficiency. These results demonstrate a role for TCF7L1 in repressing <i>LGR5</i> gene expression to govern the spheroid formation potential of CRC cells.https://www.mdpi.com/2073-4425/14/2/481colorectal cancerLGR5spheroidsT-cell factorsTCF7L1WNT |
| spellingShingle | Carli M. King Olivia M. Marx Wei Ding Walter A. Koltun Gregory S. Yochum TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells Genes colorectal cancer LGR5 spheroids T-cell factors TCF7L1 WNT |
| title | TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells |
| title_full | TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells |
| title_fullStr | TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells |
| title_full_unstemmed | TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells |
| title_short | TCF7L1 Regulates <i>LGR5</i> Expression in Colorectal Cancer Cells |
| title_sort | tcf7l1 regulates i lgr5 i expression in colorectal cancer cells |
| topic | colorectal cancer LGR5 spheroids T-cell factors TCF7L1 WNT |
| url | https://www.mdpi.com/2073-4425/14/2/481 |
| work_keys_str_mv | AT carlimking tcf7l1regulatesilgr5iexpressionincolorectalcancercells AT oliviammarx tcf7l1regulatesilgr5iexpressionincolorectalcancercells AT weiding tcf7l1regulatesilgr5iexpressionincolorectalcancercells AT walterakoltun tcf7l1regulatesilgr5iexpressionincolorectalcancercells AT gregorysyochum tcf7l1regulatesilgr5iexpressionincolorectalcancercells |