Toward the Clinical Development and Validation of a Thy1-Targeted Ultrasound Contrast Agent for the Early Detection of Pancreatic Ductal Adenocarcinoma
| Autor: | Iman Daryaei, Emmanuelle J. Meuillet, Rakesh Bam, Sanjiv S. Gambhir, Evan C. Unger, Jose G. Vilches-Moure, Amelie M. Lutz, Edmund R. Marinelli, Lotfi Abou-Elkacem, Ramasamy Paulmurugan |
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| Rok vydání: | 2020 |
| Předmět: |
Stromal cell
Contrast Media Adenocarcinoma urologic and male genital diseases Article 030218 nuclear medicine & medical imaging Flow cytometry law.invention 03 medical and health sciences Mice 0302 clinical medicine In vivo Confocal microscopy law medicine Animals Humans Radiology Nuclear Medicine and imaging Cell Lineage Early Detection of Cancer Alexa Fluor Ultrasonography Microbubbles medicine.diagnostic_test Chemistry Reproducibility of Results General Medicine In vitro Mice Inbred C57BL Pancreatic Neoplasms Cancer research Thy-1 Antigens Molecular imaging 030217 neurology & neurosurgery Ex vivo |
| Zdroj: | Invest Radiol |
| ISSN: | 1536-0210 |
| Popis: | OBJECTIVES: Early detection of Pancreatic Ductal Adenocarcinoma (PDAC) represents the most significant step towards the treatment of this aggressive lethal disease. Previously, we engineered a pre-clinical Thy1-targeted microbubble (MB(Thy1)) contrast agent that specifically recognizes Thy1 antigen overexpressed in the vasculature of murine PDAC tissues by ultrasound (US) imaging. In this study, we adopted a single-chain variable fragment (scFv) site-specific bioconjugation approach to construct clinically translatable MB(Thy1-scFv) and test for its efficacy in vivo in murine PDAC imaging and functionally evaluated the binding specificity of scFv ligand to human Thy1 in patient PDAC tissues ex vivo. MATERIALS AND METHODS: We recombinantly expressed the Thy1-scFv with a carboxy-terminus cysteine residue to facilitate its thioether conjugation to the PEGylated MBs presenting with maleimide functional groups. After the scFv-MB conjugations, we tested binding activity of the MB(Thy1-scFv) to MS1 cells over-expressing human Thy1 (MS1(Thy1)) under liquid shear stress conditions in vitro using a flow chamber set up at 0.6 mL/minute flow rate, corresponding to a wall shear stress rate of 100 seconds(−1), similar to that in tumor capillaries. For in vivo Thy1 US molecular imaging, MB(Thy1-scFv) were tested in the transgenic mouse model (C57BL/6J - Pdx1-Cre(tg/+); KRas(LSL-G12D/+;) Ink4a/Arf(−/−)) of PDAC, and in control mice (C57BL/6J) with L-arginine-induced pancreatitis or normal pancreas. To facilitate its clinical feasibility, we further produced Thy1-scFv without the bacterial fusion tags and confirmed its recognition of human Thy1 in cell lines by flow cytometry and patient PDAC frozen tissue sections of different clinical grades by immunofluorescence staining. RESULTS: Under shear stress flow conditions in vitro, MB(Thy1-scFv) bound to MS1(Thy1) cells at significantly higher numbers (3.0 ± 0.8 MB/cell; P |
| Databáze: | OpenAIRE |
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