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An Inducible, Large-Intestine-Specific Transgenic Mouse Model for Colitis and Colitis-Induced Colon Cancer Research

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Abstract

Background

Animal models are an important tool to understand intestinal biology. Our laboratory previously generated C57BL/6-Tg(Car1-cre)5Flt transgenic mice (CAC) with large-intestine-specific Cre recombinase (Cre) expression as a model to study colon health.

Aim

To expand the utility of the CAC mouse model by determining the impact of chemically induced colitis on CAC transgene expression.

Methods

CAC mice were crossed to Rosa reporter mice (Rosa26R flox/flox) with a lox-STOP-lox signal controlling β-galactosidase (βgal) expression and then further crossed with ApcCKO/CKO mice in some experiments to delete Apc alleles (ApcΔ580). Initially, 8-week-old CACTg/WT;Rosa26R flox/WT ;Apc Δ580/WT mice were treated with dextran sulfate sodium (DSS) in drinking water (5 days, 0, 0.65, 1.35, or 2.0 %). Colon tissue damage and βgal labeling were analyzed 10 day after stopping DSS. Next, 8-week-old CACTg/WT;Rosa26Rflox/flox mice were treated with 0 or 1.35 % DSS, and colonic βgal labeling was assessed at 30 day post-DSS treatment. Finally, 10-week-old CACTg/WT;Apc Δ580/WT mice were treated with DSS (0 or 2 %) for 5 days and colonic tumors were analyzed at 20 weeks.

Results

CACTg/WT;Rosa26R flox/WT ;Apc Δ580/WT mice had a DSS dose-dependent increase in colon epithelial damage that correlated with increased epithelial βgal labeling at 10 days (r 2 = 0.9, β = 0.75). The βgal labeling in CACTg/WT;Rosa26Rflox/flox mice colon remained high at 30 days, especially in the crypts of the healed ulcer. DSS also increased colon tumor incidence and multiplicity in CACTg/WT;Apc Δ580/WT mice.

Conclusions

DSS-mediated epithelial damage induces a persistent, Cre-mediated recombination of floxed alleles in CAC mice. This enables the examination of gene function in colon epithelium during experimental colitis and colitis-induced colon cancer.

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Abbreviations

APC:

Adenomatous polyposis coli

CA1:

Carbonic anhydrase I

CDX2:

Caudal-type homeobox 2

DSS:

Dextran sulfate sodium

WT:

Wild-type

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Acknowledgments

The work was supported by awards from the American Institute for Cancer Research (Award # 09A098, JCF) and the National Cancer Institute (NCI) (CA156240, JCF), a pilot grant from the Purdue University Center for Cancer Research, an NCI-designated Basic Science Cancer Center (P30 CA02316, JCF), fellowship support from the Purdue Interdisciplinary Cancer Prevention Internship Program from National Institutes of Health (R25 CA128770, FW, MLD), and an Abbott Laboratories Pathology Research Fellowship (RLJ).

Authors contributions

JCF contributed to conception and design; FW and RLJ developed the methodology; FW, RLJ, and MLD contributed to acquisition of data; FW, RLJ, and JCF analyzed and interpreted data; JCF and PWS lend administrative, technical, or material support; and JCF supervised the study.

Author information

Author notes

  1. Robert L. Johnson

    Present address: Eli Lilly and Company, Lilly Corporate Center, 355 East Merrill St., Indianapolis, IN, 46225, USA

  2. Marsha L. DeSmet

    Present address: Department of Dermatology, Indiana University School of Medicine, 550 N. University Blvd., Suite 3240, Indianapolis, IN, 46202, USA

  3. Paul W. Snyder

    Present address: EPL, Inc., 45600 Terminal Dr. # 100, Sterling, VA, 20166, USA

Authors and Affiliations

  1. Department of Nutrition Science, Purdue University, 700 West State St., West Lafayette, IN, 47907-2059, USA

    Fa Wang & James C. Fleet

  2. Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, 47907, USA

    Robert L. Johnson & Paul W. Snyder

  3. Purdue University Center for Cancer Research, Purdue University, West Lafayette, IN, 47907, USA

    Paul W. Snyder & James C. Fleet

  4. Purdue University Interdisciplinary Life Sciences Ph.D. Training Program, Purdue University, West Lafayette, IN, 47907, USA

    Marsha L. DeSmet

Authors
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Corresponding author

Correspondence to James C. Fleet.

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Conflict of interest

Dr. Fleet is on the Scientific Advisory Board for Innophos, Inc. Ms. Fa Wang and Dr. DeSmet have no conflicts to report. Dr. Johnson is currently an employee of Eli Lilly and Company. Dr. Synder is currently an employee of EPL, Inc.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of Purdue University Animal Care and Use Committee at which the studies were conducted.

Human rights

This article does not contain any studies with human participants performed by any of the authors.

Additional information

Fa Wang and Robert L. Johnson have contributed equally to the generation of this manuscript.

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Supplemental Figure S1

βgal expression in extra colonic tissues (spleen, lung, liver and kidney) was not induced by DSS treatment. Tissues were harvested, prepared, and βgal activity was detected as described for the colon. Blue staining represents βgal enzymatic activity. (A) Negative control: mice lacking the Cre-recombinase transgene. (B) βgal activity in CACTg/WT;Rosa26R flox/flox transgenic mice without DSS treatment. (C) βgal activity in CACTg/WT; Rosa26R flox/flox mice 10 days after completing a 5 day course of 1.35 % DSS. (TIFF 1136 kb)

Supplemental Figure S2

Transgene expression is increased in regenerating crypts and is sustained after healing in proximal colon. (A, B) Representative images of βgal expression level in the proximal colon of a CACTg/WT;Rosa26R flox/flox mouse treated with vehicle under 2X (A) and 20X (B) magnification. Crypt base labeling is highlighted with arrow head. (C, D) Images of βgal labeling in a CACTg/WT;Rosa26R flox/flox mouse 10 days after the end of treatment with 1.35 % DSS under 2X (C) and 20X (D) magnification. (E, F) Images of βgal labeling in a CACTg/WT;Rosa26R flox/flox mouse 30 days after the end of treatment with 1.35 % DSS under 2X (E) and 20X (F) magnification. In (C, D, E, F), Arrow = βgal positive crypts undergoing regeneration. Arrow head = β-gal positive crypts with normal phenotype adjacent to a healing area. All the images were generated using an Aperio ScanScope digital slide scanner. (TIFF 11243 kb)

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Wang, F., Johnson, R.L., Snyder, P.W. et al. An Inducible, Large-Intestine-Specific Transgenic Mouse Model for Colitis and Colitis-Induced Colon Cancer Research. Dig Dis Sci 61, 1069–1079 (2016). https://doi.org/10.1007/s10620-015-3971-7

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  • DOI: https://doi.org/10.1007/s10620-015-3971-7

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