Thy1-Targeted Microbubbles for Ultrasound Molecular Imaging of Pancreatic Ductal Adenocarcinoma

• Published in: Clinical cancer research 2018-04, Vol.24 (7), p.1574-1585
• Main Authors: Abou-Elkacem, Lotfi, Wang, Huaijun, Chowdhury, Sayan M, Kimura, Richard H, Bachawal, Sunitha V, Gambhir, Sanjiv S, Tian, Lu, Willmann, Jürgen K
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Animals; Binding; Carcinoma, Pancreatic Ductal - metabolism; Carcinoma, Pancreatic Ductal - pathology; Contrast Media - metabolism; Cytometry; Design engineering; Disease Models, Animal; Experimental design; Female; Flow cytometry; Fluid flow; Humans; Imaging; Immunofluorescence; Mice; Mice, Nude; Microbubbles; Molecular Imaging - methods; Pancreas; Pancreas - metabolism; Pancreas - pathology; Pancreatic Neoplasms; Pancreatic Neoplasms - metabolism; Pancreatic Neoplasms - pathology; Pancreatitis; Shear stress; Thy-1 Antigens - metabolism; Transgenic mice; Ultrasonic imaging; Ultrasonography - methods; Ultrasound; Xenografts; Xenotransplantation; Yeast
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-17-2057

To engineer a dual human and murine Thy1-binding single-chain-antibody ligand (Thy1-scFv) for contrast microbubble-enhanced ultrasound molecular imaging of pancreatic ductal adenocarcinoma (PDAC). Thy1-scFv were engineered using yeast-surface-display techniques. Binding to soluble human and murine Thy1 and to Thy1-expressing cells was assessed by flow cytometry. Thy1-scFv was then attached to gas-filled microbubbles to create MB Thy1 binding of MB to Thy1-expressing cells was evaluated under flow shear stress conditions in flow-chamber experiments. MB and MB were used as negative controls. All microbubble types were tested in both orthotopic human PDAC xenografts and transgenic PDAC mice Thy1-scFv had a of 3.4 ± 0.36 nmol/L for human and 9.2 ± 1.7 nmol/L for murine Thy1 and showed binding to both soluble and cellularly expressed Thy1. MB was attached to Thy1 with high affinity compared with negative control microbubbles ( < 0.01) as assessed by flow cytometry. Similarly, flow-chamber studies showed significantly ( < 0.01) higher binding of MB (3.0 ± 0.81 MB/cell) to Thy1-expressing cells than MB (0.57 ± 0.53) and MB (0.43 ± 0.53). ultrasound molecular imaging using MB demonstrated significantly higher signal ( < 0.01) in both orthotopic (5.32 ± 1.59 a.u.) and transgenic PDAC (5.68 ± 2.5 a.u.) mice compared with chronic pancreatitis (0.84 ± 0.6 a.u.) and normal pancreas (0.67 ± 0.71 a.u.). immunofluorescence confirmed significantly ( < 0.01) increased Thy1 expression in PDAC compared with chronic pancreatitis and normal pancreas tissue. A dual human and murine Thy1-binding scFv was designed to generate contrast microbubbles to allow PDAC detection with ultrasound. .