Objectives Establishing adequate resection margins and lymphatic mapping are crucial for the prognosis of oral cancer patients. Novel targeted imaging modalities are needed, enabling pre- and intraoperative detection of tumour cells, in combination with improved post-surgical examination by the pathologist. The urokinase-receptor (uPAR) is overexpressed in head and neck cancer, where it is associated with tumour progression and metastasis.
Material and methods To determine suitability of uPAR for molecular imaging of oral cancer surgery, human head and neck tumours were sectioned and stained for uPAR to evaluate the expression pattern compared to normal mucosa. Furthermore, metastatic oral squamous carcinoma cell line OSC-19 was used for targeting uPAR in in vivo mouse models. Using anti-uPAR antibody ATN-658, equipped with a multimodal label, the in vivo specificity was investigated and the optimal dose and time-window were evaluated.
Results All human oral cancer tissues expressed uPAR in epithelial and stromal cells. Hybrid ATN-658 clearly visualized tongue tumours in mice using either NIRF or SPECT imaging. Mean fluorescent TBRs over time were 4.3 ± 0.7 with the specific tracer versus 1.7 ± 0.1 with a control antibody. A significant difference in TBRs could be seen between 1 nmol (150 μg) and 0.34 nmol (50 μg) dose groups (n = 4, p < 0.05). Co-expression between BLI, GFP and the NIR fluorescent signals were seen in the tongue tumour, whereas human cytokeratin staining confirmed presence of malignant cells in the positive cervical lymph nodes.
Conclusion This study shows the applicability of an uPAR specific multimodal tracer in an oral cancer model, combining SPECT with intraoperative guidance.

Additional Metadata
Keywords Fluorescence, Head and neck, Hybrid, Imaging, Intraoperative, Near-infrared, Oral cancer, Real-time, SPECT, Surgery
Persistent URL dx.doi.org/10.1016/j.oraloncology.2016.12.026, hdl.handle.net/1765/95297
Journal Oral Oncology
Grant This work was funded by the European Commission 7th Framework Programme; grant id h2020/644373 - Preclinical Intra-Operative Image-Guided Surgery and Post-Operative Radiotherapy of Tumours (PRISAR), This work was funded by the European Commission 7th Framework Programme; grant id erc/323105 - Integrating cancer detection and SURgery Via molecular Imaging (SURVIVE)
Citation
Boonstra, M.C, van Driel, P.B.A.A, Keereweer, S, Prevoo, H.A.J.M., Stammes, M.A, Baart, V.M., … Sier, C.F.M. (2017). Preclinical uPAR-targeted multimodal imaging of locoregional oral cancer. Oral Oncology, 66, 1–8. doi:10.1016/j.oraloncology.2016.12.026