Article Figures & Data
Figures
- FIGURE 1.
Near-infrared fluorescence imaging of cell death. Imaging in vivo of untreated and treated EL4 (A), Colo-205 (B), and Eμ-myc (C) tumors. Images are overlays of bright-field images and 800 nm-channel fluorescence signals, acquired 24 h after C2Am-AF750 administration. AnxV = annexin-V.
- FIGURE 2.
Quantification of tumor mean fluorescence intensity (MFI) for untreated (open bars) and treated (filled bars) EL4 (A), Colo-205 (B), and Eμ-myc (C) tumors. Data for Eμ-myc model was acquired ex vivo, due to skin pigmentation artifacts. (Bottom) Correlation of C2Am whole-tumor MFIs with corresponding CC3 staining, measured in sections of excised EL4 (A), Colo-205 (B), and Eμ-myc (C) tumors. ○ = drug-treated (48 h after treatment); ● = untreated. **P < 0.01, ***P < 0.001, n = 3/group, values are mean ± SD. AnxV = annexin-V.
- FIGURE 3.
Maps of CC3 staining (left), C2Am-AF750 fluorescence (middle), and correlation (right) of fluorescence intensities of regions of interest (grids indicated on the left), with staining for CC3 in same regions of interest. Tumors were excised 24 h after C2Am-AF750 administration and 48 h after drug treatment. The correlation coefficients (R) of the linear fits to the data (A–C, right) are shown. Arrows in A (left) indicate decellularized regions of tissue. Sections are from EL4 (A), Colo-205 (B), and Eμ-myc (C) tumors. MFI = mean fluorescence intensity.
- FIGURE 4.
(A) Binding of 99mTc-C2Am to EL4 cells. Labeling of drug-treated and untreated cells is expressed as percentage of total 99mTc activity retained by cell pellets. **P < 0.01, ***P < 0.001, n = 3/group, values are mean ± SD. (B) Retention of 99mTc-C2Am in tumors (top) and spleens (bottom) from EL4 tumor–bearing mice, in untreated (△) and drug-treated (▲) animals, at indicated times after probe administration. Tissue retention is expressed as percentage injected dose per gram (%ID/g) of tissue. *P < 0.05, **P < 0.01, n = 3/group, 2-way ANOVA, with Bonferroni posttest correction, was used for group comparisons. Values are mean ± SD.
- FIGURE 5.
SPECT imaging of cell death in vivo in Colo-205 tumors. Imaging of 111In-labeled C2Am was performed 2 h after probe administration and 24 h after drug treatment. (A) SPECT/CT fusion images of representative untreated Colo-205 tumor–bearing mouse (A, left) and 5-FU–treated animal (A, right), 2 h after administration of 111In-C2Am. Tumor location is indicated by arrowheads. (B) Tumor retention (percentage injected dose [%ID]/mL) (upper) and CC3 staining (lower) in untreated and 5-FU–treated tumors. *P < 0.05, n = 3−4 tumors/group.
- FIGURE 6.
SPECT imaging of cell death in vivo in Eμ-myc tumors. Imaging of 99mTc-labeled C2Am was performed 2 h after probe administration, and 24 h after drug treatment. (A) SPECT/CT fusion images of representative Eμ-myc mice before (left) and after (right) cyclophosphamide treatment. Tumors in neck, axillary region, and chest cavity were visible (arrowheads, top) and in axial sections across cervical (1, middle) and axillary (2, bottom) planes. Red circles in B correspond to percentage injected dose [%ID]/mL retention values for tumors of animals shown in A. CC3 staining (B, bottom) in untreated and drug-treated tumors. *P < 0.05, ***P < 0.0001, n = 6−7 tumors/group (A); n = 6−13 tumors/group (B). AT = after treatment; BT = before treatment. Supplemental materials provide statistical analysis. Three-dimensional rendering of SPECT data are shown in Supplemental Video 1.
Tables
- TABLE 1
Biodistribution of 99mTc-C2Am in Tumor-Bearing EL4 Mice and 111In-C2Am in Tumor-Bearing Colo-205 Mice, 24 Hours After Etoposide and 5-FU Treatment, Respectively
2 h 24 h Tissue (%ID/g) Mean SD Mean SD Tumor-bearing EL4 mice Muscle 0.39 0.05 0.19 0.02 Blood 1.02 0.08 0.20 0.01 Tumor 1.74 0.39 1.41 0.21 Spleen 3.30 0.47 2.34 0.29 Liver 6.17 0.98 5.14 0.90 Kidney 194.2 45.9 157.4 57.0 Tumor-to-blood 1.71 0.40 6.96 1.08 Tumor-to-muscle 4.5 1.15 7.3 1.40 Tumor-bearing Colo-205 mice Muscle 0.32 0.07 0.18 0.02 Blood 0.50 0.22 0.13 0.02 Tumor 0.69 0.09 0.72 0.10 Spleen 0.92 0.17 1.18 0.20 Liver 2.01 0.34 2.17 0.53 Kidney 310 12 275 22 Tumor-to-blood 1.38 0.65 5.43 0.96 Tumor-to-muscle 2.16 0.48 4.09 0.88 %ID/g = percentage injected dose per gram.
n = 3/group.
Supplemental Data
Files in this Data Supplement:
