A multimodal molecular imaging approach targeting urokinase plasminogen activator receptor for the diagnosis, resection and surveillance of urothelial cell carcinoma

Victor M Baart,Geertje Van Der Horst,Marion M Deken,Shadhvi S Bhairosingh,Timo Schomann,Vincent Q Sier,Maaike H Van Der Mark,Luisa Iamele,Hugo De Jonge,Massimo Resnati,Andrew P Mazar,Rob C.M Pelger,Gabriel Van Der Pluijm,Peter J.K Kuppen,Alexander L Vahrmeijer,Cornelis F.M Sier

doi:10.1016/j.ejca.2021.01.001

Victor M Baart, Geertje Van Der Horst + Show 14 more

Open Access

https://doi.org/10.1016/j.ejca.2021.01.001

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Abstract

With a 5-year recurrence rate of 30–78%, urothelial cell carcinoma (UCC) rates amongst the highest of all solid malignancies. Consequently, after transurethral resection, patients are subjugated to life-long endoscopic surveillance. A multimodal near-infrared (NIR) fluorescence-based imaging strategy can improve diagnosis, resection and surveillance, hence increasing quality of life. MethodsExpression of urokinase plasminogen activator receptor (uPAR) and epithelial cell adhesion molecule (EpCAM) are determined on paraffin-embedded human UCC using immunohistochemistry and on UCC cell lines by flow cytometry. MNPR-101, a humanised monoclonal antibody targeting uPAR is conjugated to IRDye800CW and binding is validated in vitro using surface plasmon resonance and cell-based binding assays. In vivo NIR fluorescence and photoacoustic three-dimensional (3D) imaging are performed with subcutaneously growing human UM-UC-3luc2 cells in BALB/c-nude mice. The translational potential is confirmed in a metastasising UM-UC-3luc2 orthotopic mouse model. Infliximab-IRDye800CW and rituximab-IRDye800CW are used as controls. ResultsUCCs show prominent uPAR expression at the tumour-stroma interface and EpCAM on epithelial cells. uPAR and EpCAM are expressed by 6/7 and 4/7 UCC cell lines, respectively. In vitro, MNPR-101-IRDye800CW has a picomolar affinity for domain 2-3 of uPAR. In vivo fluorescence imaging with MNPR-101-IRDye800CW, specifically delineates both subcutaneous and orthotopic tumours with tumour-to-background ratios reaching as high as 6.8, differing significantly from controls (p < 0.0001). Photoacoustic 3D in depth imaging confirms the homogenous distribution of MNPR-101-IRDye800CW through the tumour. ConclusionsMNPR-101-IRDye800CW is suitable for multimodal imaging of UCC, awaiting clinical translation.

Highlights

Despite advances in detection, treatment and surveillance of urothelial cell carcinoma (UCC), there has been no major improvement in overall prognosis over the past 30 years, with nearly 200,000 patients still succumbing annually [1,2]
UCC causes a high burden of disease, where patients could benefit from improved transurethral resection (TUR) and tumourfree resection margins
One-third of cases had no epithelial cell adhesion molecule (EpCAM) expression. urokinase plasminogen activator receptor (uPAR) staining was most prominent at the tumour-stroma interface localising towards cancer cells and tumourassociated stromal cells, including macrophages (CD68 positive) and cancer-associated fibroblasts (Fig. 1B)

Summary

Introduction

Treatment and surveillance of urothelial cell carcinoma (UCC), there has been no major improvement in overall prognosis over the past 30 years, with nearly 200,000 patients still succumbing annually [1,2]. UCC represents two sequential entities: non-muscle invasive bladder cancer (NMIBC), where malignant cells are constrained to the epithelial layer, and muscle-invasive bladder cancer (MIBC) wherein the tumour invades surrounding subepithelial tissue [3]. The majority of UCC cases (75e85%) are NMIBC and are marked by a high 5-year recurrence rate of 30e78% and 7e40% chance of progression to MIBC disease after transurethral resection (TUR). UCC causes a high burden of disease, where patients could benefit from improved TUR and tumourfree resection margins

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