Anti-αvβ3 antibody guided three-step pretargeting approach using magnetoliposomes for molecular magnetic resonance imaging of breast cancer angiogenesis

Abstract

Chenggong Yan,1 Yuankui Wu,1 Jie Feng,1 Wufan Chen,2 Xiang Liu,1 Peng Hao,1 Ruimeng Yang,3 Juan Zhang,2 Bingquan Lin,1 Yikai Xu,1 Ruiyuan Liu41Department of Medical Imaging Center, Nanfang Hospital, Southern Medical University, Guangzhou, People's Republic of China; 2School of Biomedical Engineering, Southern Medical University, Guangzhou, People's Republic of China; 3Department of Radiology, the First Municipal People's Hospital of Guangzhou, Guangzhou, People's Republic of China; 4School of Pharmaceutical Science, Southern Medical University, People's Republic of ChinaPurpose: Pretargeting of biomarkers with nanoparticles in molecular imaging is promising to improve diagnostic specificity and realize signal amplification, but data regarding its targeting potential in magnetic resonance (MR) imaging are limited. The purpose of this study was to evaluate the tumor angiogenesis targeting efficacy of the anti-αvβ3 antibody guided three-step pretargeting approach with magnetoliposomes.Methods: Polyethylene glycol-modified and superparamagnetic iron oxide-encapsulated magnetoliposomes with and without biotin were synthesized and characterized. The cytotoxicity of both probes was evaluated using the methyl thiazdyl tetrazolium assay, and their cellular uptake by mouse macrophage was visualized using Prussian blue staining. Three-step pretargeting MR imaging was performed on MDA-MB-435S breast cancer-bearing mice by intravenous administration of biotinylated anti-αvβ3 monoclonal antibodies (first step), followed by avidin and streptavidin (second step), and by biotinylated magnetoliposomes or magnetoliposomes in the targeted or nontargeted group, respectively (third step). The specificity of αvβ3 targeting was assessed by histologic examinations.Results: The developed magnetoliposomes were superparamagnetic and biocompatible as confirmed by cell toxicity assay. The liposomal bilayer and polyethylene glycol modification protected Fe3O4 cores from uptake by macrophage cells. MR imaging by three-step pretargeting resulted in a greater signal enhancement along the tumor periphery, occupying 7.0% of the tumor area, compared with 2.0% enhancement of the nontargeted group (P < 0.05). Histologic analysis demonstrated the targeted magnetoliposomes colocalized with neovasculature, which was responsible for the MR signal decrease.Conclusion: These results indicate that our strategy for MR imaging of αvβ3-integrin is an effective means for sensitive detection of tumor angiogenesis, and may provide a targetable nanodelivery system for anticancer drugs.Keywords: pretargeting, contrast agents, superparamagnetic iron oxide nanoparticles, avidin-biotin