Near-Infrared Fluorescence Imaging of Pancreatic Cancer Using a Fluorescently Labelled Anti-CEA Nanobody Probe: A Preclinical Study

• Published in: Biomolecules (Basel, Switzerland) Switzerland), 2023-03, Vol.13 (4), p.618
• Main Authors: van Manen, Labrinus, de Muynck, Lizzie D A N, Baart, Victor M, Bhairosingh, Shadhvi, Debie, Pieterjan, Vahrmeijer, Alexander L, Hernot, Sophie, Mieog, J Sven D
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Animals; antibody; Antigens; Cameras; Cancer; Cancer surgery; Cancer therapies; Carcinoembryonic Antigen; Carcinoma, Pancreatic Ductal - diagnostic imaging; CEA (Oncology); Cell culture; Cell Line, Tumor; Chromatography; Coloring Agents; Communication; Dyes; Feasibility studies; Flow cytometry; Fluorescence; Humans; I.R. radiation; Investigations; Medical imaging equipment; Medical research; Methylene blue; Mice; Monoclonal antibodies; Nanobodies; nanobody; near-infrared; Oncology, Experimental; Optical Imaging - methods; Pancreatic cancer; pancreatic ductal adenocarcinoma; Pancreatic Neoplasms; Pancreatic Neoplasms - diagnostic imaging; Pancreatic Neoplasms - pathology; Surface active agents; Surgery; Tumors; Zwitterions
• ISSN: 2218-273X; 2218-273X
• DOI: 10.3390/biom13040618

Molecular fluorescence-guided surgery using near-infrared light has the potential to improve the rate of complete resection of cancer. Typically, monoclonal antibodies are being used as targeting moieties, however smaller fragments, such as single-domain antibodies (i.e., Nanobodies ) improve tumor specificity and enable tracer injection on the same day as surgery. In this study, the feasibility of a carcinoembryonic antigen-targeting Nanobody (NbCEA5) conjugated to two zwitterionic dyes (ZW800-1 Forte [ZW800F] and ZW800-1) for visualization of pancreatic ductal adenocarcinoma (PDAC) was investigated. After site-specific conjugation of NbCEA5 to the zwitterionic dyes, binding specificity was evaluated on human PDAC cell lines with flow cytometry. A dose escalation study was performed for both NbCEA5-ZW800F and NbCEA5-ZW800-1 in mice with subcutaneously implanted pancreatic tumors. Fluorescence imaging was performed up to 24 h after intravenous injection. Furthermore, the optimal dose for NbCEA5-ZW800-1 was injected in mice with orthotopically implanted pancreatic tumors. A dose-escalation study showed superior mean fluorescence intensities for NbCEA5-ZW800-1 compared to NbCEA5-ZW800F. In the orthotopic tumor models, NbCEA5-ZW800-1 accumulated specifically in pancreatic tumors with a mean in vivo tumor-to-background ratio of 2.4 (SD = 0.23). This study demonstrated the feasibility and potential advantages of using a CEA-targeted Nanobody conjugated to ZW800-1 for intraoperative PDAC imaging.