Targeting Diagnosis of High-Risk Papillary Thyroid Carcinoma Using Ultrasound Contrast Agent With the BRAF V600E Mutation: An Experimental Study

• Published in: Journal of ultrasound in medicine Vol. 41; no. 11; pp. 2789 - 2802
• Main Authors: Xie, Fang, Yan, Lin, Li, Yi-Ming, Lan, Yu, Xiao, Jing, Zhang, Ming-Bo, Jin, Zhuang, Zhang, Ying, Tian, Xiao-Qi, Zhu, Ya-Qiong, Li, Zhi-Ping, Luo, Yu-Kun
• Format: Journal Article
• Language: English
• Published: England 01.11.2022
• Subjects: contrast enhanced ultrasound; BRAFV600E mutation; papillary thyroid carcinoma; targeted nanobubbles
• ISSN: 0278-4297; 1550-9613
• DOI: 10.1002/jum.15967

High-risk papillary thyroid carcinoma (PTC) patients with BRAF mutation have lymph node and distant metastasis and poor prognosis. Therefore, this study aims to develop a targeted ultrasound contrast agent for the BRAF mutation to screen high-risk PTC at early stage. The targeted lipid nanobubbles carrying BRAF antibody were prepared using thin film hydration-sonication and avidin-biotin binding methods. The physicochemical properties and stability of the targeted nanobubbles were detected by transmission electron microscopy, atomic force microscopy, and confocal laser scanning microscopy. The target binding abilities of the targeted nanobubbles in the PTC cells (B-CPAP) overexpressed mutant BRAF were evaluated by immunofluorescence staining, quantitative real-time polymerase chain reaction, western blot, and fluorescence microscopy. After PTC tumor models overexpressed mutant BRAF were established, the enhanced images of targeted lipid nanobubbles and untargeted lipid nanobubbles on PTC tumors in nude mice were observed using contrast-enhanced ultrasound imaging. The targeted lipid nanobubbles revealed uniform, round morphology, and good stability with a nanoscale size. Besides, BRAF monoclonal antibody was observed to be combined on the surface of lipid nanobubbles. Furthermore, the targeted nanobubbles had a good targeting diagnosis ability in PTC cells with BRAF overexpression. Moreover, the targeted nanobubbles had better ultrasound enhancement and peak intensity of the time-intensity curve (P < .001) in PTC tumors with BRAF overexpression as compared to the untargeted lipid nanobubbles. The targeted lipid nanobubbles carrying BRAF antibody could be regarded as a potential targeted ultrasound contrast agent for the diagnosis of high-risk PTC.