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Mice deficient in Muc4 are resistant to experimental colitis and colitis-associated colorectal cancer

Oncogene volume 35, pages 2645–2654 (2016)Cite this article

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Abstract

MUC4, a large transmembrane mucin normally expressed in the small and large intestine, is differentially expressed during inflammatory and malignant conditions of the colon. However, the expression pattern and the role of MUC4 in colitis and colorectal cancer (CRC) are inconclusive. Therefore, the aim of this study was to understand the role of Muc4 during inflammatory and malignant conditions of the colon. Here, we generated Muc4−/− mice and addressed its role in colitis and colitis-associated CRC using dextran sodium sulfate (DSS) and azoxymethane (AOM)-DSS experimental models, respectively. Muc4−/− mice were viable, fertile with no apparent defects. Muc4−/− mice displayed increased resistance to DSS-induced colitis compared with wild-type (WT) littermates that was evaluated by survival rate, body weight loss, diarrhea and fecal blood score, and histological score. Reduced infiltration of inflammatory cells, that is, CD3+ lymphocytes and F4/80+ macrophages was observed in the inflamed mucosa along with reduction in the mRNA levels of inflammatory cytokines interleukin (IL)-1β and tumor necrosis factor (TNF)-α and anti-microbial genes Lysozyme M and SLPI in the colon of Muc4−/− mice compared with WT littermates. Compensatory upregulation of Muc2 and Muc3 mucins under basal and DSS treatment conditions partly explains the resistance observed in Muc4−/− mice. Accordingly, Muc4−/− mice exhibited significantly reduced tumor burden compared with WT mice assessed in a colitis-induced tumor model using AOM/DSS. An increased percentage of Ki67+ nuclei was observed in the tumors from WT compared with Muc4−/− mice suggesting Muc4 to be critical in intestinal cell proliferation during tumorigenesis. Taken together, we conclusively demonstrate for the first time the role of Muc4 in driving intestinal inflammation and inflammation-associated tumorigenesis using a novel Muc4−/− mouse model.

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Abbreviations

AOM:

azoxymethane

CD:

Crohn’s disease

CRC:

colorectal cancer

DSS:

dextran sodium sulfate

IBD:

inflammatory bowel disease

IL:

interleukin

TNF:

tumor necrosis factor

UC:

ulcerative colitis

WT:

wild type

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Acknowledgements

We thank Don Harms and Rolen Quadros of the mouse genome-engineering core at the University of Nebraska Medical Center (UNMC) for the electroporation, microinjection and generation of chimera for Muc4 knockout mice. We thank UNMC tissue sciences facility for processing of tissues used in the study. We also thank Dr Imayavaramban Lakshmanan and Kavita Mallya for technical assistance. The work in the manuscript, in parts, was supported by the grants from the National Institutes of Health (RO1 CA 78590, RO1 CA183459, UO1CA111294, SPORE P50CA127297, TMEN U54CA163120 and R03 CA167342).

Author contributions

SD, SR and SKB: study concept and design; SD and SR: acquisition of data; CBG: generation of Muc4−/− mice; YS: histopathological analysis; LMS: statistics; SD, SR, HKR and SKB: analysis and interpretation of data; SD and SKB: wrote the manuscript.

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Author notes

  1. S Das

    Present address: 7Current Address: Department of Medicine, Karolinska Universitetssjukhuset, Solna, Stockholm 171 76, Sweden.,

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA

    S Das, S Rachagani & S K Batra

  2. Department of Pathology, University of Nebraska Medical Center, Omaha, NE, USA

    Y Sheinin

  3. Department of Biostatistics, University of Nebraska Medical Center, Omaha, NE, USA

    L M Smith

  4. Department of Genetics, Mouse Genome Engineering Core Facility, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE, USA

    C B Gurumurthy

  5. Center for Digestive Disorders, Boston University School of Medicine, Boston, MA, USA

    H K Roy

  6. Buffett Cancer Center, Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA

    S K Batra

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Correspondence to S K Batra.

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Das, S., Rachagani, S., Sheinin, Y. et al. Mice deficient in Muc4 are resistant to experimental colitis and colitis-associated colorectal cancer. Oncogene 35, 2645–2654 (2016). https://doi.org/10.1038/onc.2015.327

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