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In vivo micro-CT imaging of untreated and irradiated orthotopic glioblastoma xenografts in mice: capabilities, limitations and a comparison with bioluminescence imaging

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Abstract

Small animal imaging is of increasing relevance in biomedical research. Studies systematically assessing the diagnostic accuracy of contrast-enhanced in vivo micro-CT of orthotopic glioma xenografts in mice do not exist. NOD/SCID/γc−/− mice (n = 27) underwent intracerebral implantation of 2.5 × 106 GFP-Luciferase-transduced U87MG cells. Mice underwent bioluminescence imaging (BLI) to detect tumor growth and afterwards repeated contrast-enhanced (300 µl Iomeprol i.v.) micro-CT imaging (80 kV, 75 µAs, 360° rotation, 1,000 projections, 33 s scan time, resolution 40 × 40 × 53 µm, 0.5 Gy/scan). Presence of tumors, tumor diameter and tumor volume in micro-CT were rated by two independent readers. Results were compared with histological analyses. Six mice with tumors confirmed by micro-CT received fractionated irradiation (3 × 5 Gy every other day) using the micro-CT (5 mm pencil beam geometry). Repeated micro-CT scans were tolerated well. Tumor engraftment rate was 74 % (n = 20). In micro-CT, mean tumor volume was 30 ± 33 mm3, and the smallest detectable tumor measured 360 × 620 µm. The inter-rater agreement (n = 51 micro-CT scans) for the item tumor yes/no was excellent (Spearman-Rho = 0.862, p < 0.001). Sensitivity and specificity of micro-CT were 0.95 and 0.71, respectively (PPV = 0.91, NPV = 0.83). BLI on day 21 after tumor implantation had a sensitivity and specificity of 0.90 and 1.0, respectively (PPV = 1.0, NPV = 0.5). Maximum tumor diameter and volume in micro-CT and histology correlated excellently (tumor diameter: 0.929, p < 0.001; tumor volume: 0.969, p < 0.001, n = 17). Irradiated animals showed a large central tumor necrosis. Longitudinal contrast enhanced micro-CT imaging of brain tumor growth in live mice is feasible at high sensitivity levels and with excellent inter-rater agreement and allows visualization of radiation effects.

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Acknowledgments

We are indebted to Dr. M. Müller for assistance in preparation of this manuscript.

Conflict of interest

None of the authors of the above manuscript have declared any conflict of interest.

Funding

The acquisition of the micro-CT (Yxlon Y. Fox) was funded by the Federal Ministry of Education and Research and the Land Baden-Württemberg (HBFG Grant#125-648). This work was funded by the Deutsche Forschungsgemeinschaft (Grants FL 880/1-1, GI 771/1-1. GL 236/9-1 and WE 2063/9-1), the Bundesministerium für Bildung und Forschung (Federal Ministry of Education and Research, grant BMBF 01EZ1130), the Klaus Tschira Stiftung (Grant 00.211.2012) and the Bundesamt für Strahlenschutz (Federal Office for Radiation Protection, Grant BfS 3608S04001) for the establishment of the endowed professorship Medizinische Strahlenphysik/Strahlenschutz (Medical Radiation Physics/Radiation Protection).

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    Authors and Affiliations

    1. Department of Neuroradiology, Medical Faculty Mannheim, University, Medical Center Mannheim, Heidelberg University, 68167, Mannheim, Germany

      Stefanie Kirschner, Máté E. Maros & Hans U. Kerl

    2. Medical Radiation Physics/Radiation Protection, Department of Radiation Oncology, Medical Faculty Mannheim, University Medical Center Mannheim, Heidelberg University, 68167, Mannheim, Germany

      Manuela C. Felix & Gerhard Glatting

    3. Department of Radiation Oncology, Medical Faculty Mannheim, University Medical Center Mannheim, Heidelberg University, 68167, Mannheim, Germany

      Linda Hartmann, Miriam Bierbaum, Frederik Wenz & Frank A. Giordano

    4. Department of Veterinary Clinical Sciences, Small Animal Clinic, Justus-Liebig-University, 35392, Giessen, Germany

      Martin Kramer

    5. Clinic of Diagnostic and Interventional Neuroradiology, University Hospital of the RWTH Aachen, Pauwelsstr. 30, 52074, Aachen, Germany

      Marc A. Brockmann

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    1. Stefanie Kirschner
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    Correspondence to Marc A. Brockmann.

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    Frank A. Giordano and Marc A. Brockmann these authors have contributed equally.

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    Kirschner, S., Felix, M.C., Hartmann, L. et al. In vivo micro-CT imaging of untreated and irradiated orthotopic glioblastoma xenografts in mice: capabilities, limitations and a comparison with bioluminescence imaging. J Neurooncol 122, 245–254 (2015). https://doi.org/10.1007/s11060-014-1708-7

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    • DOI: https://doi.org/10.1007/s11060-014-1708-7

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