Detection of Experimental Colorectal Peritoneal Metastases by a Novel PDGFRβ-Targeting Nanobody.

Abstract

Simple SummaryColorectal cancer can metastasize to multiple distant sites. Metastases growing within the peritoneal cavity cause a high degree of morbidity and are associated with very poor survival. Moreover, peritoneal metastases are difficult to detect using conventional imaging methods. Consequently, peritoneal metastases are generally under-diagnosed and their response to therapy is difficult to assess. An extensive molecular and cellular analysis of colorectal peritoneal metastases revealed that these lesions express very high levels of specific markers that could serve as targets for imaging-based diagnosis and treatment. In the present report, we explore the potential value of one such marker, PDGFRB, to serve as a target for peritoneal metastasis detection by molecular imaging. Therefore, we generated a PDGFRB-binding llama nanobody and demonstrate its utility in detecting peritoneal metastases in mice. The clinical development of PDGFRB-targeting tracers may help to improve the diagnosis of peritoneal metastases and the clinical management of this highly aggressive disease entity.Peritoneal metastases in colorectal cancer (CRC) belong to Consensus Molecular Subtype 4 (CMS4) and are associated with poor prognosis. Conventional imaging modalities, such as Computed Tomography (CT) and Fluorodeoxyglucose-Positron Emission Tomography (FDG-PET), perform very poorly in the detection of peritoneal metastases. However, the stroma-rich nature of these lesions provides a basis for developing molecular imaging strategies. In this study, conducted from 2019 to 2021, we aimed to generate a Platelet-Derived Growth Factor Receptor beta (PDGFRB)-binding molecular imaging tracer for the detection of CMS4 CRC, including peritoneal metastases. The expression of PDGFRB mRNA discriminated CMS4 from CMS1-3 (AUROC = 0.86 (95% CI 0.85–0.88)) and was associated with poor relapse-free survival. PDGFRB mRNA and protein levels were very high in all human peritoneal metastases examined (n = 66). Therefore, we generated a PDGFRB-targeting llama nanobody (VHH1E12). Biotin-labelled VHH1E12 bound to immobilized human and mouse PDGFRB with high affinity (EC50 human PDGFRB = 7 nM; EC50 murine PDGFRB = 0.8 nM), and to PDGFRB-expressing HEK293 cells grown in vitro. A pharmacokinetic analysis of IRDye-800CW-conjugated VHH1E12 in mice showed that the plasma half-life was 6 min. IRDye-800CW-conjugated VHH1E12 specifically accumulated in experimentally induced colorectal cancer peritoneal metastases in mice. A tissue analysis subsequently demonstrated co-localization of the nanobody with PDGFRB expression in the tumour stroma. Our results demonstrate the potential value of PDGFRB-targeted molecular imaging as a novel strategy for the non-invasive detection of CMS4 CRC, in particular, peritoneal metastases.