Antibody-based PET of uPA/uPAR signaling with broad applicability for cancer imaging

• Published in: Oncotarget 2016-11, Vol.7 (45), p.73912-73924
• Main Authors: Yang, Dongzhi, Severin, Gregory W, Dougherty, Casey A, Lombardi, Rachel, Chen, Daiqin, Van Dort, Marcian E, Barnhart, Todd E, Ross, Brian D, Mazar, Andrew P, Hong, Hao
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal; Biomarkers; Cell Line, Tumor; Disease Models, Animal; Fluorescent Antibody Technique; Heterografts; Humans; Mice; Molecular Imaging - methods; Neoplasms - diagnostic imaging; Neoplasms - metabolism; Neoplasms - pathology; Positron-Emission Tomography - methods; Protein Binding; Radiopharmaceuticals; Receptors, Urokinase Plasminogen Activator - metabolism; Research Paper; Tissue Distribution; Urokinase-Type Plasminogen Activator - metabolism
• ISSN: 1949-2553; 1949-2553
• DOI: 10.18632/oncotarget.12528

Mounting evidence suggests that the urokinase plasminogen activator (uPA) and its receptor (uPAR) play a central role in tumor progression. The goal of this study was to develop an 89Zr-labeled, antibody-based positron emission tomography (PET) tracer for quantitative imaging of the uPA/uPAR system. An anti-uPA monoclonal antibody (ATN-291) was conjugated with a deferoxamine (Df) derivative and subsequently labeled with 89Zr. Flow cytometry, microscopy studies, and competitive binding assays were conducted to validate the binding specificity of Df-ATN-291 against uPA. PET imaging with 89Zr-Df-ATN-291 was carried out in different tumors with distinct expression levels of uPA. Biodistribution, histology examination, and Western blotting were performed to correlate tumor uptake with uPA or uPAR expression. ATN-291 retained uPA binding affinity and specificity after Df conjugation. 89Zr-labeling of ATN-291 was achieved in good radiochemical yield and high specific activity. Serial PET imaging demonstrated that, in most tumors studied (except uPA- LNCaP), the uptake of 89Zr-Df-ATN-291 was higher compared to major organs at 120 h post-injection, providing excellent tumor contrast. The tumor-to-muscle ratio of 89Zr-Df-ATN-291 in U87MG was as high as 45.2 ± 9.0 at 120 h p.i. In vivo uPA specificity of 89Zr-Df-ATN-291 was confirmed by successful pharmacological blocking of tumor uptake with ATN-291 in U87MG tumors. Although the detailed mechanisms behind in vivo 89Zr-Df-ATN-291 tumor uptake remained to be further elucidated, quantitative PET imaging with 89Zr-Df-ATN-291 in tumors can facilitate oncologists to adopt more relevant cancer treatment planning.