Wnt signaling is one of the most common pathways linked to carcinogenesis in the intestine. The transcription factors that mediate Wnt signals are members of the Lymphoid Enhancer Factor/T Cell Factor (LEF/TCF) family. This four-member family is complex in that there are full-length activating forms, truncated dominant negative forms, and alternatively spliced isoforms that modify Wnt target gene recognition. TCF-1 and TCF-4 are the two family members expressed in the epithelial crypt compartment of the intestine. Normally, TCF-1 is expressed as a dominant negative isoform and it counteracts the action of TCF-4 which is expressed as a Wnt-mediating, active isoform. This pattern changes in cancer because TCF-1 expression switches to a full-length, Wnt mediating form. We have discovered a second DNA binding domain in TCF-1 and TCF-4 isoforms called the C-clamp. We have shown that the C-clamp is important for regulating cell proliferation of colon cancer cells. We have also identified Wnts that trigger TCF-1 export in colon cancer cells. Export of TCF-1 leaves the C-clamp form of TCF-4 in the nucleus to mediate aberrant Wnt signaling.
Three aims are proposed for this project. First, the extracellular Wnts and intracellular components that direct TCF-1 nuclear export will be defined (Aim1). Second, the biological impact of export will be defined in colon cancer cells and colon cancer initiating 3D cultures (Aim2). Third, the DNA binding specificities and target gene selection of C-clamp forms of TCF-1 and TCF-4 will be defined through genome-wide binding assays (ChIP-Seq), high-throughput protein binding microarrays, and in vitro biochemical analysis (Aim3). The overall goal is to define the mechanisms behind changes in TCF expression in colon carcinogenesis, and to define how extracellular Wnts and DNA binding specificities of TCFs influence aberrant signaling in colon cancer cells with stabilized ?-catenin.

Public Health Relevance

In colon cancer, Wnt signaling is an overactive and strong signal for tumor initiation and progression. Wnt signaling is mediated by the TCF transcription factors and we have discovered their activities and expression to be different in colon cancer versus normal colon tissue. This project investigates basic mechanisms of TCF localization and gene targeting and asks how differences in these attributes contribute to the cancer cell phenotype.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA108697-07
Application #
8248742
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Mietz, Judy
Project Start
2004-07-01
Project End
2016-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
7
Fiscal Year
2012
Total Cost
$287,159
Indirect Cost
$91,298
Name
University of California Irvine
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Sprowl-Tanio, Stephanie; Habowski, Amber N; Pate, Kira T et al. (2016) Lactate/pyruvate transporter MCT-1 is a direct Wnt target that confers sensitivity to 3-bromopyruvate in colon cancer. Cancer Metab 4:20
Vuong, Linh M; Chellappa, Karthikeyani; Dhahbi, Joseph M et al. (2015) Differential Effects of Hepatocyte Nuclear Factor 4α Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells. Mol Cell Biol 35:3471-90
Ehsan, Seema M; Welch-Reardon, Katrina M; Waterman, Marian L et al. (2014) A three-dimensional in vitro model of tumor cell intravasation. Integr Biol (Camb) 6:603-10
MacDonald, Bryan T; Hien, Annie; Zhang, Xinjun et al. (2014) Disulfide bond requirements for active Wnt ligands. J Biol Chem 289:18122-36
Pate, Kira T; Stringari, Chiara; Sprowl-Tanio, Stephanie et al. (2014) Wnt signaling directs a metabolic program of glycolysis and angiogenesis in colon cancer. EMBO J 33:1454-73
Hoverter, Nate P; Zeller, Michael D; McQuade, Miriam M et al. (2014) The TCF C-clamp DNA binding domain expands the Wnt transcriptome via alternative target recognition. Nucleic Acids Res 42:13615-32
Chodaparambil, Jayanth V; Pate, Kira T; Hepler, Margretta R D et al. (2014) Molecular functions of the TLE tetramerization domain in Wnt target gene repression. EMBO J 33:719-31
Wu, Beibei; Piloto, Sarah; Zeng, Weihua et al. (2013) Ring Finger Protein 14 is a new regulator of TCF/ýý-catenin-mediated transcription and colon cancer cell survival. EMBO Rep 14:347-55
Konstorum, Anna; Sprowl, Stephanie A; Waterman, Marian L et al. (2013) Predicting mechanism of biphasic growth factor action on tumor growth using a multi-species model with feedback control. J Coupled Syst Multiscale Dyn 1:459-467
Hoverter, Nate P; Ting, Ju-Hui; Sundaresh, Suman et al. (2012) A WNT/p21 circuit directed by the C-clamp, a sequence-specific DNA binding domain in TCFs. Mol Cell Biol 32:3648-62

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