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EGFR-mediated macrophage activation promotes colitis-associated tumorigenesis

Oncogene volume 36, pages 3807–3819 (2017)Cite this article

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Abstract

Epidermal growth factor receptor (EGFR) signaling is a known mediator of colorectal carcinogenesis. Studies have focused on the role of EGFR signaling in epithelial cells, although the exact nature of the role of EGFR in colorectal carcinogenesis remains a topic of debate. Here, we present evidence that EGFR signaling in myeloid cells, specifically macrophages, is critical for colon tumorigenesis in the azoxymethane–dextran sodium sulfate (AOM-DSS) model of colitis-associated carcinogenesis (CAC). In a human tissue microarray, colonic macrophages demonstrated robust EGFR activation in the pre-cancerous stages of colitis and dysplasia. Utilizing the AOM-DSS model, mice with a myeloid-specific deletion of Egfr had significantly decreased tumor multiplicity and burden, protection from high-grade dysplasia and significantly reduced colitis. Intriguingly, mice with gastrointestinal epithelial cell-specific Egfr deletion demonstrated no differences in tumorigenesis in the AOM-DSS model. The alterations in tumorigenesis in myeloid-specific Egfr knockout mice were accompanied by decreased macrophage, neutrophil and T-cell infiltration. Pro-tumorigenic M2 macrophage activation was diminished in myeloid-specific Egfr-deficient mice, as marked by decreased Arg1 and Il10 mRNA expression and decreased interleukin (IL)-4, IL10 and IL-13 protein levels. Surprisingly, diminished M1 macrophage activation was also detectable, as marked by significantly reduced Nos2 and Il1b mRNA levels and decreased interferon (IFN)-γ, tumor necrosis factor (TNF)-α and IL-1β protein levels. The alterations in M1 and M2 macrophage activation were confirmed in bone marrow-derived macrophages from mice with the myeloid-specific Egfr knockout. The combined effect of restrained M1 and M2 macrophage activation resulted in decreased production of pro-angiogenic factors, CXCL1 and vascular endothelial growth factor (VEGF), and reduced CD31+ blood vessels, which likely contributed to protection from tumorigenesis. These data reveal that EGFR signaling in macrophages, but not in colonic epithelial cells, has a significant role in CAC. EGFR signaling in macrophages may prove to be an effective biomarker of CAC or target for chemoprevention in patients with inflammatory bowel disease.

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Acknowledgements

CD3, CD45R and CD31 staining was performed by the Vanderbilt University Medical Center Translational Pathology Shared Resource supported by NIH grant P30CA068485 and the Vanderbilt Mouse Metabolic Phenotyping Center, supported by NIH grant U24DK059637. Slide imaging and quantification was performed in the Digital Histology Shared Resource (www.mc.vanderbilt.edu/dhsr). This work was funded by NIH grants R01DK053620, R01AT004821, R01CA190612, P01CA116087 and P01CA028842 (KTW), Veterans Affairs Merit Review grant I01BX001453 (KTW), the Thomas F Frist Sr Endowment (KTW) and the Vanderbilt Center for Mucosal Inflammation and Cancer (KTW). The human TMA was supported by NIH grant P50CA095103 and the DDRC Tissue Morphology Subcore (NIH grant P30DK058404). DMH was supported by T32GM008554 and F31DK10715. LAC was supported by Veterans Affairs Career Development Award 1IK2BX002126.

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Authors and Affiliations

  1. Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, TN, USA

    D M Hardbower, M K Washington & K T Wilson

  2. Division of Gastroenterology, Department of Medicine, Hepatology and Nutrition, Vanderbilt University Medical Center, Nashville, TN, USA

    D M Hardbower, L A Coburn, M Asim, K Singh, J C Sierra, D P Barry, A P Gobert, M B Piazuelo & K T Wilson

  3. Center for Mucosal Inflammation and Cancer, Vanderbilt University Medical Center,, Nashville, TN, USA

    L A Coburn, A P Gobert, M B Piazuelo & K T Wilson

  4. Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN, USA

    L A Coburn & K T Wilson

  5. Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    K T Wilson

  6. Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center,, Nashville, TN, USA

    K T Wilson

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  1. D M Hardbower
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Hardbower, D., Coburn, L., Asim, M. et al. EGFR-mediated macrophage activation promotes colitis-associated tumorigenesis. Oncogene 36, 3807–3819 (2017). https://doi.org/10.1038/onc.2017.23

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