Near-Infrared Imaging of Colonic Adenomas In Vivo Using Orthotopic Human Organoids for Early Cancer Detection

SIMPLE SUMMARY: Colorectal cancer is a leading cause of worldwide cancer-related morbidity and mortality. Conventional white light colonoscopy is routinely used to identify and remove colonic polyps. However, premalignant lesions that are flat and subtle in appearance are often missed. Cells alter in molecular expression well in advance of morphological changes. cMet is highly overexpressed by colonic adenomas. We aim to implant patient-derived organoids in the colon of immunocompromised mice to demonstrate a model system for evaluating the specific uptake of a near-infrared labeled peptide ligand. Human adenoma and normal colonoids were imaged in vivo using a small animal endoscope. A high target-to-background ratio was observed ~1 h post-injection followed by clearance after ~24 h. The mean intensity measured was significantly greater for the adenoma versus normal colonoids. These results demonstrate promise for a targeted imaging approach using fluorescently labeled peptides to identify precancerous lesions that would otherwise be missed using white light illumination alone. ABSTRACT: Colorectal cancer is a leading cause of cancer-related morbidity and mortality worldwide. Premalignant lesions that are flat and subtle in morphology are often missed in conventional colonoscopies. Patient-derived adenoma colonoids with high and low cMet expression and normal colonoids were implanted orthotopically in the colon of immunocompromised mice to serve as a preclinical model system. A peptide specific for cMet was labeled with IRDye800, a near-infrared (NIR) fluorophore. This peptide was administered intravenously, and in vivo imaging was performed using a small animal fluorescence endoscope. Quantified intensities showed a peak target-to-background ratio at ~1 h after intravenous peptide injection, and the signal cleared by ~24 h. The peptide was stable in serum with a half-life of 3.6 h. Co-staining of adenoma and normal colonoids showed a high correlation between peptide and anti-cMet antibody. A human-specific cytokeratin stain verified the presence of human tissues implanted among surrounding normal mouse colonic mucosa. Peptide biodistribution was consistent with rapid renal clearance. No signs of acute toxicity were found on either animal necropsy or serum hematology and chemistries. Human colonoids provide a clinically relevant preclinical model to evaluate the specific uptake of a NIR peptide to detect premalignant colonic lesions in vivo.