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In vivo MRI virtual colonography in a mouse model of colon cancer

Abstract

We have developed a reliable noninvasive method for monitoring colonic tumors and mucosal inflammation in a mouse model of colon cancer using magnetic resonance colonography (MRC). After a mild cleansing enema, the colon is filled with Fluorinert, a perfluorinated liquid that does not produce a proton MR signal. The mouse is placed in a dedicated volume MR receiver coil, and high-resolution images are acquired in three planes. The Fluorinert enema distends the mouse colon, creating an artifact-free black homogeneous background, allowing clear delineation of the inflamed colonic wall and visualization of luminal tumors in various stages of development. A gadolinium-based contrast agent can be administered i.v. to the animal to detect mural inflammation or tumor vascularity. This technique is useful for serial monitoring of the effects of preventive or therapeutic strategies on tumor development without killing the animal or requiring invasive endoscopies. The animal preparation and imaging can be completed in 1.5 h.

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Figure 1: Sagittal, coronal and transverse views of the 3D T1w images acquired with different enema media.
Figure 2: Tools setup.
Figure 3: Planning of the 3D slab.
Figure 4: MR images of the mouse colon of healthy and tumor-bearing mice.
Figure 5: Multiplanar reformat package generates transverse and sagittal views of the colon.
Figure 6: Calculation of the volumes of individual tumors with ImageJ.
Figure 7: The digital subtraction of the precontrast and postcontrast T1w images enhances the tumor vascularity, removes the signal from residual stools and improves the sensitivity of the method.
Figure 8

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References

  1. Jemal, A., Siegel, R., Xu, J. & Ward, E. Cancer statistics, 2010. CA Cancer J. Clin. 60, 277–300 (2010).

    Article  Google Scholar 

  2. Becker, C., Fantini, M.C. & Neurath, M.F. High-resolution colonoscopy in live mice. Nat. Protoc. 1, 2900–2904 (2006).

    Article  CAS  Google Scholar 

  3. Becker, C. et al. In vivo imaging of colitis and colon cancer development in mice using high-resolution chromoendoscopy. Gut 54, 950–954 (2005).

    Article  CAS  Google Scholar 

  4. Durkee, B.Y., Weichert, J.P. & Halberg, R.B. Small animal micro-CT colonography. Methods 50, 36–41 (2010).

    Article  CAS  Google Scholar 

  5. Pickhardt, P.J. et al. Microcomputed tomography colonography for polyp detection in an in vivo mouse tumor model. Proc. Natl. Acad. Sci. 102, 3419–3422 (2005).

    Article  CAS  Google Scholar 

  6. Boll, H. et al. Double-contrast micro-CT colonoscopy in live mice. Int. J. Colorectal Dis. 26, 721–727 (2011).

    Article  Google Scholar 

  7. Krupnick, A.S. et al. Quantitative monitoring of mouse lung tumors by magnetic resonance imaging. Nat. Protoc. 7, 128–142 (2012).

    Article  CAS  Google Scholar 

  8. Amundson, S.A. et al. Induction of gene expression as a monitor of exposure to ionizing radiation. Radiat. Res. 156, 657–661 (2001).

    Article  CAS  Google Scholar 

  9. Saar, B. et al. Prospective study on bright lumen magnetic resonance colonography in comparison with conventional colonoscopy. Br. J. Radiol. 80, 235–241 (2007).

    Article  CAS  Google Scholar 

  10. Ajaj, W. et al. Dark lumen magnetic resonance colonography: comparison with conventional colonoscopy for the detection of colorectal pathology. Gut 52, 1738–1743 (2003).

    Article  CAS  Google Scholar 

  11. Zijta, F.M. et al. Feasibility of using automated insufflated carbon dioxide (CO2) for luminal distension in 3.0T MR colonography. Eur. J. Radiol. 81, 1128–1133 (2012).

    Article  CAS  Google Scholar 

  12. Keeling, A.N. et al. Intravenous, contrast-enhanced MR colonography using air as endoluminal contrast agent: impact on colorectal polyp detection. Eur. J. Radiol. 81, 31–38 (2012).

    Article  Google Scholar 

  13. Graser, A. Magnetic Resonance colonography. Radiol. Clin. N. Am. 51, 113–120 (2013).

    Article  Google Scholar 

  14. Lauenstein, T.C., Goehde, S.C. & Debatin, J.F. Fecal tagging: MR colonography without colonic cleansing. Abdom. Imaging 4, 410–417 (2002).

    Article  Google Scholar 

  15. Hensley, H.H., Chang, W.C. & Clapper, M.L. Detection and volume determination of colonic tumors in Min mice by magnetic resonance micro-imaging. Magn. Reson. Med. 52, 524–529 (2004).

    Article  Google Scholar 

  16. Melgar, S., Gillberg, P.G., Hockings, P.D. & Olsson, L.E. High-throughput magnetic resonance imaging in murine colonic inflammation. Biochem. Biophys. Res. Commun. 355, 1102–1107 (2007).

    Article  CAS  Google Scholar 

  17. Larsson, A.E. et al. Magnetic resonance imaging of experimental mouse colitis and association with inflammatory activity. Inflamm. Bowel Dis. 12, 478–485 (2006).

    Article  Google Scholar 

  18. Herborn, C.U. et al. Dark lumen magnetic resonance colonography in a rodent polyp model: initial experience and demonstration of feasibility. Invest. Radiol. 39, 723–727 (2004).

    Article  Google Scholar 

  19. Breynaert, C. et al. Unique gene expression and MR T2 relaxometry patterns define chronic murine dextran sodium sulphate colitis as a model for connective tissue changes in human Crohn's disease. PLoS ONE 8, e68876 (2013).

    Article  CAS  Google Scholar 

  20. Young, M.R. et al. Monitoring of tumor promotion and progression in a mouse model of inflammation-induced colon cancer with magnetic resonance colonography. Neoplasia 11, 237–246 (2009).

    Article  CAS  Google Scholar 

  21. Tanaka, T. et al. A novel inflammation-related mouse colon carcinogenesis model induced by azoxymethane and dextran sodium sulfate. Cancer Sci. 94, 965–973 (2003).

    Article  CAS  Google Scholar 

  22. Neufert, C., Becker, C. & Neurath, M.F. An inducible mouse model of colon carcinogenesis for the analysis of sporadic and inflammation-driven tumor progression. Nat. Protoc. 2, 1998–2004 (2007).

    Article  CAS  Google Scholar 

  23. Saud, S. et al. Chemopreventive activity of plant flavonoid isorhamnetin in colorectal cancer is mediated by oncogenic Src and β-catenin. Cancer Res. 73, 5473–5484 (2013).

    Article  CAS  Google Scholar 

  24. Miller, S.J. et al. Multimodal imaging of growth and rapamycin-induced regression of colonic adenomas in Apc mutation–dependent mouse. Transl. Oncol. 5, 313–320 (2012).

    Article  Google Scholar 

  25. Quarles, C.C. et al. Functional colonography of Min mice using dark lumen dynamic contrast-enhanced MRI. Magn. Reson. Med. 60, 718–726 (2008).

    Article  Google Scholar 

  26. Flaim, S.F. Pharmacokinetics and side effects of perfluorocarbon-based blood substitutes. Artif. Cells Blood Substit. Immobil. Biotechnol. 22, 1043–1054 (1994).

    Article  CAS  Google Scholar 

  27. Chu, S.J. et al. Systemic administration of FC-77 dampens ischemia-reperfusion-induced acute lung injury in rats. Inflammation 36, 1383–1392 (2013).

    Article  CAS  Google Scholar 

  28. Chang, H. et al. Intravascular FC-77 attenuates phorbol myristate acetate-induced acute lung injury in isolated rat lungs. Crit. Care Med. 36, 1222–1229 (2008).

    Article  CAS  Google Scholar 

  29. Zhu, Y.B. et al. Partial liquid ventilation decreases tissue and serum tumor necrosis factor-α concentrations in acute lung injury model of immature piglet induced by oleic acid. Chin. Med. J. (Engl) 125, 123–128 (2012).

    CAS  Google Scholar 

  30. Hirayama, Y. et al. Partial liquid ventilation with FC-77 suppresses the release of lipid mediators in rat acute lung injury model. Crit. Care Med. 32, 2085–2089 (2004).

    Article  CAS  Google Scholar 

  31. Maevsky, E., Ivanitsky, G. & Bogdanova, L. Clinical results of Perftoran application: present and future. Artif. Cells Blood Substit. Immobil. Biotechnol. 33, 37–46 (2005).

    Article  CAS  Google Scholar 

  32. Kaneda, M.M., Caruthers, S., Lanza, G.M. & Wickline, S.A. Perfluorocarbon nanoemulsions for quantitative molecular imaging and targeted therapeutics. Ann. Biomed. Eng. 37, 1922–1933 (2009).

    Article  Google Scholar 

  33. Giraudeau, C. et al. 19F molecular MR imaging for detection of brain tumor angiogenesis: in vivo validation using targeted PFOB nanoparticles. Angiogenesis 16, 171–179 (2013).

    Article  CAS  Google Scholar 

  34. Liao, A.H. et al. Evaluation of 18F-labeled targeted perfluorocarbon-filled albumin microbubbles as a probe for micro-US and micro-PET in tumor-bearing mice. Ultrasonics 53, 320–327 (2013).

    Article  CAS  Google Scholar 

  35. Turkbey, B. et al. Is apparent diffusion coefficient associated with clinical risk scores for prostate cancers that are visible on 3-T MR images? Radiology 258, 488–495 (2011).

    Article  Google Scholar 

  36. Turkbey, B. et al. Documenting the location of systematic transrectal ultrasound-guided prostate biopsies: correlation with multi-parametric MRI. Cancer Imaging 11, 31–36 (2011).

    Article  Google Scholar 

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Acknowledgements

This project has been funded in whole or in part with federal funds from the National Cancer Institute, US National Institutes of Health, under contract HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products or organizations imply endorsement by the US Government.

Author information

Authors and Affiliations

Authors

Contributions

L.V.I. performed the MRC experiments and prepared the manuscript; M.B. designed the imaging protocol, assembled the imaging tools, reconstructed and described the fly-through movie and edited the manuscript; M.R.Y. designed the animal study and edited the manuscript; L.A.R. provided animal support; J.L.T. advised and discussed on the imaging methods; J.D.K. supervised the project and edited the manuscript; P.L.C. developed the project concept and edited the manuscript.

Corresponding author

Correspondence to Lilia V Ileva.

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Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 MRI equipment setup.

(a) Philips Achieva 3.0T clinical scanner; (b) Mouse solenoid receiver coil with plastic chamber; (c) Mouse imaging bed; and (d) Syringe pumps for contrast media and enema infusion.

Supplementary information

Supplementary Figure 1

MRI equipment setup. (PDF 270 kb)

Fly-through movie.

The 3D image data allow reconstruction of a virtual colonoscopy fly-through movie that enables tumor search inside the walls of the colon. Refer to Box 2 for advice on how to do this. (MOV 9631 kb)

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Ileva, L., Bernardo, M., Young, M. et al. In vivo MRI virtual colonography in a mouse model of colon cancer. Nat Protoc 9, 2682–2692 (2014). https://doi.org/10.1038/nprot.2014.178

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