CD146-targeted immunoPET and NIRF Imaging of Hepatocellular Carcinoma with a Dual-Labeled Monoclonal Antibody.
CD146-targeted immunoPET and NIRF Imaging of Hepatocellular Carcinoma with a Dual-Labeled Monoclonal Antibody.
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作者：
Hernandez R;Sun H;England CG;Valdovinos HF;Ehlerding EB;Barnhart TE;Yang Y;Cai W
CD146 的过度表达与肝细胞癌 (HCC) 患者的侵袭性、复发率和较差的总体生存率相关。在这项研究中，我们着手开发一种 CD146 靶向探针，用于 HCC 的高对比度无创体内正电子发射断层扫描 (PET) 和近红外荧光 (NIRF) 成像。 YY146 是一种抗 CD146 单克隆抗体，被用作靶向分子，我们将两性离子近红外荧光 (NIRF) 染料 ZW800-1 和螯合剂去铁胺 (Df) 与其结合。这使得能够用 89Zr 标记 Df-YY146-ZW800 并随后使用 PET 和 NIRF 成像进行检测，所有这些都不会影响抗体结合特性。采用表达高水平 (HepG2) 和低水平 (Huh7) CD146 的两种 HCC 细胞系在无胸腺裸鼠中产生皮下 (s.c) 和原位异种移植物。将 89Zr-Df-YY146-ZW800 静脉注射到荷瘤小鼠体内后进行的连续 PET 和 NIRF 成像显示，HepG2 肿瘤中示踪剂的显着且持续的摄取，在注射后 72 小时达到每克注射剂量百分比 (%ID/g; n=4) 的 31.65 ± 7.15 百分比峰值。由于这种明显的积累，PET 和 NIRF 都能成功勾画肿瘤，这有助于荧光图像引导原位 HepG2 肿瘤的切除，尽管肝脏背景相对较高。 CD146 阴性 Huh7 和 CD146 阻断的 HepG2 肿瘤表现出显着较低的 89Zr-Df-YY146-ZW800 积聚（注射后 72 小时分别为 6.1 ± 0.5 和 8.1 ± 1.0 %ID/g；n=4），证明了示踪剂在体内的 CD146 特异性。离体生物分布和免疫荧光染色证实了成像数据的准确性以及示踪剂摄取与原位 CD146 表达的相关性。总体而言，89Zr-Df-YY146-ZW800 作为 PET/NIRF 成像剂表现出优异的特性，包括对表达 CD146 的 HCC 具有高体内亲和力和特异性。使用双标记 YY146 的 CD146 靶向分子成像在肝脏恶性肿瘤的早期检测、预测和图像引导手术切除方面具有巨大潜力。
Overexpression of CD146 has been correlated with aggressiveness, recurrence rate, and poor overall survival in hepatocellular carcinoma (HCC) patients. In this study, we set out to develop a CD146-targeting probe for high-contrast noninvasive in vivo positron emission tomography (PET) and near-infrared fluorescence (NIRF) imaging of HCCs. YY146, an anti-CD146 monoclonal antibody, was employed as a targeting molecule to which we conjugated the zwitterionic near-infrared fluorescence (NIRF) dye ZW800-1 and the chelator deferoxamine (Df). This enabled labeling of Df-YY146-ZW800 with 89Zr and its subsequent detection using PET and NIRF imaging, all without compromising antibody binding properties. Two HCC cell lines expressing high (HepG2) and low (Huh7) levels of CD146 were employed to generate subcutaneous (s.c.) and orthotopic xenografts in athymic nude mice. Sequential PET and NIRF imaging performed after intravenous injection of 89Zr-Df-YY146-ZW800 into tumor-bearing mice unveiled prominent and persistent uptake of the tracer in HepG2 tumors that peaked at 31.65 ± 7.15 percentage of injected dose per gram (%ID/g; n=4) 72 h post-injection. Owing to such marked accumulation, tumor delineation was successful by both PET and NIRF, which facilitated the fluorescence image-guided resection of orthotopic HepG2 tumors, despite the relatively high liver background. CD146-negative Huh7 and CD146-blocked HepG2 tumors exhibited significantly lower 89Zr-Df-YY146-ZW800 accretion (6.1 ± 0.5 and 8.1 ± 1.0 %ID/g at 72 h p.i., respectively; n=4), demonstrating the CD146-specificity of the tracer in vivo. Ex vivo biodistribution and immunofluorescent staining corroborated the accuracy of the imaging data and correlated tracer uptake with in situ CD146 expression. Overall, 89Zr-Df-YY146-ZW800 showed excellent properties as a PET/NIRF imaging agent, including high in vivo affinity and specificity for CD146-expressing HCC. CD146-targeted molecular imaging using dual-labeled YY146 has great potential for early detection, prognostication, and image-guided surgical resection of liver malignancies.