Over the past 20 years our understanding of the molecular etiology of colon cancer has expanded dramatically. Despite this, colon cancer remains a significant health problem in the US. One of the barriers to progress in the field is the lack of well-characterized animal models that recapitulate the etiology of human colon cancer. While we have learned a great deal from cancers resulting from various chemically induced- or genetically programmed-rodent models, the cancer that develops in these models is often significantly different from human colon cancer in terms of latency, intestinal location, or molecular signature. Mechanistic and prevention focused colon cancer research requires the development of animal models that permit precisely timed, colon-specific modification of intestinal biology. Our goal for the proposed research is to use the promoter that drives large intestine-specific expression of the carbonic anhydrase 1 (CA1) gene to create a transgenic mouse with colon-epithelial cell-specific expression of Cre recombinase (Cre)-fused to a modified estrogen receptor (ER) ligand binding domain with high affinity only for the estrogen analog tamoxifen (CAC- ERT2). The ERT2 portion of the fusion gene confers taxomifen-inducibility to the transgene and permits temporally controlled deletion of floxed alleles in the mouse genome. A transgene vector has been produced and it will be used to produce mice at the Purdue Transgenic Mouse Core Facility. Transgene expression level and tissue distribution will be assessed by PCR. Inducibility of transgene function by tamoxifen will be assessed after crossing the CAC-ERT2 mouse to the ROSA26R indicator mouse. The ability of tamoxifen to induce tumor formation will be assessed in CAC-ERT2 mice crossed to mice with one or two floxed APC alleles. Additional studies will be done in the presence of colonic inflammation induced by dextran sulfate sodium (DSS). Upon completion of the project, the CAC-ERT2 mouse will be the only transgenic model that both limits Cre expression to the epithelial cells of the large intestine and permits control over when Cre functions to delete floxed alleles. This will permit colon cancer researchers to easily combine genetic mutations (i.e. with multiple floxed alleles relevant to colon cancer etiology) and initiate colon cancer at any stage of life. This will permit more careful assessment of mechanisms of carcinogenesis and improved testing of chemopreventative or therapeutic agents in adult mice. While our interests are in using this model for colon cancer, this model will also be useful for researchers interested in inflammatory conditions of the lower bowel (i.e. IBD, Crohn's disease). 1

Public Health Relevance

Colon cancer is a common form of cancer in the US and scientists. Research to develop effective treatment and prevention strategies is hampered by the lack of animal models that reproduce the features of human sporadic colon cancer. Our proposed research will create a new genetically modified mouse model that overcomes the limitations of existing animal models for colon cancer research.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21CA165240-01
Application #
8246227
Study Section
Cancer Genetics Study Section (CG)
Program Officer
Mietz, Judy
Project Start
2012-02-16
Project End
2014-01-31
Budget Start
2012-02-16
Budget End
2013-01-31
Support Year
1
Fiscal Year
2012
Total Cost
$162,728
Indirect Cost
$53,978
Name
Purdue University
Department
Nutrition
Type
Other Domestic Higher Education
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Wang, Fa; Johnson, Robert L; Snyder, Paul W et al. (2016) An Inducible, Large-Intestine-Specific Transgenic Mouse Model for Colitis and Colitis-Induced Colon Cancer Research. Dig Dis Sci 61:1069-79
DeWitt, Marsha; Johnson, Robert L; Snyder, Paul et al. (2015) The effect of 1,25 dihydroxyvitamin D3 treatment on the mRNA levels of ? catenin target genes in mice with colonic inactivation of both APC alleles. J Steroid Biochem Mol Biol 148:103-10
Fleet, James C (2014) Animal models of gastrointestinal and liver diseases. New mouse models for studying dietary prevention of colorectal cancer. Am J Physiol Gastrointest Liver Physiol 307:G249-59
Byun, Alexander J; Hung, Kenneth E; Fleet, James C et al. (2014) Colon-specific tumorigenesis in mice driven by Cre-mediated inactivation of Apc and activation of mutant Kras. Cancer Lett 347:191-5
Johnson, Robert L; Fleet, James C (2013) Animal models of colorectal cancer. Cancer Metastasis Rev 32:39-61