Research ArticleBasic Science Investigations

Claudin-4 SPECT Imaging Allows Detection of Aplastic Lesions in a Mouse Model of Breast Cancer

Michael Mosley, James Knight, Albrecht Neesse, Patrick Michl, Manuela Iezzi, Veerle Kersemans and Bart Cornelissen
Journal of Nuclear Medicine May 2015, 56 (5) 745-751; DOI: https://doi.org/10.2967/jnumed.114.152496

Article Figures & Data

Figures

  • FIGURE 1.
    FIGURE 1.

    (A) MDA-MB-468, SQ20b, and HT1080 cells were stained for claudin-4. Scale bar, 20 μm. (B) Western blot demonstrating presence or absence of claudin-4 in whole cell lysates obtained from MDA-MB-468, SQ20b, and HT1080 cells.

  • FIGURE 2.
    FIGURE 2.

    (A) MDA-MB-468 cells were exposed for 1 h at 4°C to increasing concentrations of 111In-labeled cCPE.GST or GST, and extent of cell-binding was determined. (B) Increasing amounts of unlabeled cCPE.GST were used to block binding of 111In-cCPE.GST to MDA-MB-468 cells.

  • FIGURE 3.
    FIGURE 3.

    (A) Representative SPECT/CT images of mice carrying tumor xenografts (white circles) of MDA-MB-468 (claudin-4–positive) or HT1080 (claudin-4–negative) cells, 24 h after intravenous administration of 111In-cCPE.GST or 111In-GST. 111In uptake was also observed in shoulder joints (S), liver (L), intestines (I), and knee joints (K). Coronal sections through tumor are shown. (B) Sections obtained from MDA-MB-468 or HT1080 xenograft tumors were stained using anticlaudin-4 antibodies. Scale bar, 20 μm. (C) Biodistribution results, 24 h after intravenous administration of 111In-cCPE.GST or 111In-GST. Tumor-to-blood and tumor-to-muscle ratios were calculated. Each group contained at least 3 animals. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

  • FIGURE 4.
    FIGURE 4.

    (A) Sections obtained from tumors harvested from 139-d-old BALB/neuT mouse were stained using anti–claudin-4 antibodies. Scale bar, 20 μm. (B) Representative SPECT/CT images of BALB/neuT mice, aged 139 d, bearing mammary tumors (white circles), 24 h after intravenous administration of 111In-cCPE.GST or 111In-GST. Overt tumors were clearly visible on CT images. Coronal sections through tumor are shown. (C) Volume-of-interest analysis of BALB/neuT mice bearing overt tumors 3 or 24 h after injection of 111In-cCPE.GST or 111In-GST. Each group contained at least 3 animals. *P < 0.05.

  • FIGURE 5.
    FIGURE 5.

    (A) Sections obtained from aplastic lesion or muscle harvested from 80-d-old BALB/neuT mouse or ductal tissue obtained from a wild-type BALB/c mouse were stained using anti–claudin-4 antibodies. Scale bar, 20 μm. (B) Representative SPECT/CT images of BALB/neuT mice, aged 80 d, bearing aplastic lesions (white circles), 24 h after intravenous administration of 111In-cCPE.GST or 111In-GST. Functional SPECT imaging was compared with anatomic CT imaging. Aplastic lesion could not be detected on CT images. Coronal sections through mammary fat pads are shown. (C) Volume-of-interest analysis of 80-d-old BALB/neuT mice bearing aplastic 24 h after injection of 111In-cCPE.GST or 111In-GST. Each group contained at least 3 animals. *P < 0.05.

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