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Endoscopy-guided orthotopic implantation of colorectal cancer cells results in metastatic colorectal cancer in mice

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Abstract

Advanced stage colorectal cancer (CRC) is still associated with limited prognosis. For preclinical evaluation of novel therapeutic approaches, murine models with orthotopic tumor growth and distant metastases are required. However, these models usually require surgical procedures possibly influencing tumor immunogenicity and development. The aim of this study was to establish a minimal-invasive endoscopy-based murine orthotopic model of metastatic CRC. During colonoscopy of CD-1 nude and non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice, implantation of Caco-2 and HT-29 CRC cells was performed subcutaneously (s.c.) or orthotopic into the colonic submucosa. White light endoscopy (WLE) and fluorescence endoscopy (FE) were applied for tumor detection in vivo. Ex vivo, resected tumors were examined by fluorescence reflectance imaging (FRI), histology, gelatin zymography and immunohistochemistry. In CD-1 nude mice, marked tumor growth was observed within 14 days after subcutaneous implantation while submucosal implantation failed to induce CRC after 17 weeks. In contrast, in NOD/SCID mice submucosal injection of HT-29 cells resulted in pronounced tumor growth 12 days post injectionem. Subsequently, rapid tumor expansion occurred, occupying the entire colonic circumference. Importantly, post mortem histological analyses confirmed liver metastases in 28.6 % and peritoneal metastases in 14.3 % of all mice. FRI and gelatin zymography did not detect a significantly increased matrix metalloproteinases (MMPs) expression in s.c. implanted tumors while MMP-tracer uptake was significantly enhanced in orthotopic implanted tumors. Neither s.c. nor orthotopic Caco-2 cell implantation resulted in tumor development. We successfully established an endoscopy-based model of metastatic CRC in immunodeficient mice.

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Abbreviations

CRC:

Colorectal cancer

MMP:

Matrix metalloproteinase

FRI:

Fluorescence reflectance imaging

FE:

Fluorescence endoscopy

NOD:

Non-obese diabetic

SCID:

Severe combined immunodeficiency

S.C.:

Subcutaneous

NIR:

Near infrared

MMPI:

Matrix metalloproteinases inhibitor

G:

Gauge

HE:

Hematoxylin eosin

IL:

Interleukin

TNF:

Tumor necrosis factor

ROI:

Region of interest

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Acknowledgments

We thank S. Dufentester, E. Weber, I. Hoppe, C. Möllmann and D. Reinhardt for their expert technical assistance. This work was supported by an interdisciplinary grant from the Else-Kröner-Fresenius-Stiftung (2012_A94 to PL, DB, DD and MS). P. Lenz was supported by a research fellowship from the Deutsche Forschungsgemeinschaft, collaborative research center SFB 656, Münster, Germany. We thank Faekah Gohar for proofreading the manuscript. D. Bettenworth and P. Lenz were supported by a research fellowship from the Faculty of Medicine, Westfälische Wilhelms-Universität Münster.

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Authors and Affiliations

  1. Department of Medicine B, University Hospital of Münster, Münster, Germany

    Dominik Bettenworth, Marcus M. Mücke & Dirk Domagk

  2. European Institute for Molecular Imaging, University of Münster, Münster, Germany

    Katrin Schwegmann, Andreas Faust, Michael Schäfers & Philipp Lenz

  3. Institute of Pathology Munich-North, Munich, Germany

    Christopher Poremba

  4. Cells-in-Motion Cluster of Excellence (EXC 1003 – CiM), University of Münster, Münster, Germany

    Michael Schäfers

  5. Josephs-Hospital Warendorf, Warendorf, Germany

    Dirk Domagk

  6. Institute of Palliative Care, University Hospital of Münster, Albert-Schweitzer-Campus 1, Building W30, 48149, Münster, Germany

    Philipp Lenz

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  1. Dominik Bettenworth
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Correspondence to Philipp Lenz.

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Dominik Bettenworth and Marcus M. Mücke authors share equal first authorship.

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Bettenworth, D., Mücke, M.M., Schwegmann, K. et al. Endoscopy-guided orthotopic implantation of colorectal cancer cells results in metastatic colorectal cancer in mice. Clin Exp Metastasis 33, 551–562 (2016). https://doi.org/10.1007/s10585-016-9797-7

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  • DOI: https://doi.org/10.1007/s10585-016-9797-7

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