Early detection of response to experimental chemotherapeutic Top216 with [18F]FLT and [18F]FDG PET in human ovary cancer xenografts in mice

Mette Munk Jensen, Kamille Dumong Erichsen, Fredrik Björkling, Jacob Madsen, Peter Buhl Jensen, Liselotte Højgaard, Maxwell Sehested, Andreas Kjær, Mette Munk Jensen, Kamille Dumong Erichsen, Fredrik Björkling, Jacob Madsen, Peter Buhl Jensen, Liselotte Højgaard, Maxwell Sehested, Andreas Kjær

Abstract

Background: 3'-Deoxy-3'-[(18)F]fluorothymidine ((18)F-FLT) is a tracer used to assess cell proliferation in vivo. The aim of the study was to use (18)F-FLT positron emission tomography (PET) to study treatment responses to a new anti-cancer compound. To do so, we studied early anti-proliferative effects of the experimental chemotherapy Top216 non-invasively by PET.

Methodology/principal findings: In vivo uptake of (18)F-FLT in human ovary cancer xenografts in mice (A2780) was studied at various time points after Top216 treatment (50 mg/kg i.v. at 0 and 48 hours) was initiated. Baseline (18)F-FLT scans were made before either Top216 (n = 7-10) or vehicle (n = 5-7) was injected and repeated after 2 and 6 hours and 1 and 5 days of treatment. A parallel study was made with 2'-deoxy-2'-[(18)F]fluoro-D-glucose ((18)F-FDG) (n = 8). Tracer uptake was quantified using small animal PET/CT. Imaging results were validated by tumor volume changes and gene-expression of Ki67 and TK1. Top216 (50 mg/kg 0 and 48 hours) inhibited the growth of the A2780 tumor compared to the control group (P<0.001). (18)F-FLT uptake decreased significantly at 2 hours (-52%; P<0.001), 6 hours (-49%; P = 0.002) and Day 1 (-47%; P<0.001) after Top216 treatment. At Day 5 (18)F-FLT uptake was comparable to uptake in the control group. Uptake of (18)F-FLT was unchanged in the control group during the experiment. In the treatment group, uptake of (18)F-FDG was significantly decreased at 6 hours (-21%; P = 0.003), Day 1 (-29%; P<0.001) and Day 5 (-19%; P = 0.05) compared to baseline.

Conclusions/significance: One injection with Top216 initiated a fast and significant decrease in cell-proliferation assessable by (18)F-FLT after 2 hours. The early reductions in tumor cell proliferation preceded changes in tumor size. Our data indicate that (18)F-FLT PET is promising for the early non-invasive assessment of chemotherapy effects in both drug development and for tailoring therapy in patients.

Conflict of interest statement

Competing Interests: The following co-authors have conflict of interests: Peter Buhl Jensen: Ownership Interests and Employment in Topotarget A/S. Maxwell Sehested: Ownership Interests and Employment in Topotarget A/S. Fredrik Björkling: Employment in Topotarget A/S. Kamille Dumong Erichsen: Employment in Topotarget A/S. All other authors have no conflict of interests. That some of the co-authors are employed by Topotarget A/S does not alter the authors' adherence to the PLoS ONE policies on sharing data and materials.

Figures

Figure 1. Chemical structure of Top216.
Figure 1. Chemical structure of Top216.
Figure 2. Schematic view of the experimental…
Figure 2. Schematic view of the experimental design.
Figure 3. Tumor growth.
Figure 3. Tumor growth.
A) The effects of Top216 on the growth of A2780 tumor xenografts. Tumor volume was determined by microCT. Mice were treated with Top216 (50 mg/kg) or vehicle at 0 and 48 hours. *) P# # #) P<0.001 vs. control. n = 15 tumors per group. B) Changes in tumor volume assessed by ratio Day 5/baseline in the control group as a function of baseline FLT uptake. n = 7 tumors. R2 = 0.61, P = 0.04.
Figure 4. Fused PET/CT images.
Figure 4. Fused PET/CT images.
A) The eight images at left are representative coronal fused PET/CT images of two mice scanned with 18F-FLT at baseline and at 6 hours and 1 and 5 days after treatment start. The images at the top show one mouse treated with Top216 and the images at the bottom show one control mouse which received vehicle. The four images at right show fused PET/CT pictures of two representative mice treated with either Top216 or vehicle and scanned at baseline and 2 hours after treatment initiation. The arrows point towards the tumors. B) Representative coronal fused PET/CT images of two mice scanned with 18F-FDG at baseline and 6 hours and 1 and 5 days after treatment start. The images at the top show one mouse treated with Top216 and the images at the bottom show one control mouse which received vehicle. The arrows point towards the tumors.
Figure 5. 18 F-FLT and 18 F-FDG…
Figure 5. 18F-FLT and 18F-FDG uptake assessed by SUVmean and SUVmax at baseline and following treatment with Top216.
Top216/vehicle treatment was initiated at 0 hours and repeated at 48 hours after the first injection. N = 5–10 tumors per group. *) P18F-FLT experiments and the two graphs at right show data from the 18F-FDG experiments.
Figure 6. Expression of Ki67 and TK1…
Figure 6. Expression of Ki67 and TK1 normalized to expression of reference gene TBP.
Data are presented as fold changes following treatment with Top216/vehicle relative to baseline levels (n = 7 tumors per group). Top216 treatment was initiated at 0 hours and repeated at 48 hours after the first injection. *) P

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