Nanobodies Targeting Mouse/Human VCAM1 for the Nuclear Imaging of Atherosclerotic Lesions
Nanobodies Targeting Mouse/Human VCAM1 for the Nuclear Imaging of Atherosclerotic Lesions
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DOI：
10.1161/circresaha.112.265140
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发表时间：
2012-03-30
影响因子：
20.1
通讯作者：
Devoogdt, Nick
中科院分区：
文献类型：
--
作者：
Broisat, Alexis;Hernot, Sophie;Devoogdt, Nick
理由：一种能够检测易损动脉粥样硬化斑块的无创工具是非常需要的。通过结合纳摩尔亲和力和快速血液清除，纳米体代表心血管分子成像的潜在放射性示踪剂。血管细胞粘附分子-1 （VCAM1）是动脉粥样硬化病变分子成像的相关靶点。目的：我们旨在生成、放射性标记和评估抗vcam1纳米体用于动脉粥样硬化病变的无创检测。方法与结果：制备10个抗VCAM1纳米体，用锝-99m放射性标记，体外筛选小鼠和人重组VCAM1蛋白和内皮细胞，体内筛选载脂蛋白e缺陷（ApoE(-/-)）小鼠。在所有实验中都使用了非靶向控制纳米体来证明特异性。所有纳米体对小鼠VCAM1均表现出纳摩尔亲和力。利用人脐静脉内皮细胞进行的流式细胞术分析显示，10个纳米体中有6个具有小鼠和人的VCAM1交叉反应性。先导化合物cAbVCAM1-5对人VCAM1具有交叉反应性，具有较高的病变控制比（4.95+/-0.85）、病变心脏比（8.30+/-1.11）和病变血液比（4.32+/-0.48）
Rationale: A noninvasive tool allowing the detection of vulnerable atherosclerotic plaques is highly needed. By combining nanomolar affinities and fast blood clearance, nanobodies represent potential radiotracers for cardiovascular molecular imaging. Vascular cell adhesion molecule-1 (VCAM1) constitutes a relevant target for molecular imaging of atherosclerotic lesions.Objective: We aimed to generate, radiolabel, and evaluate anti-VCAM1 nanobodies for noninvasive detection of atherosclerotic lesions.Methods and Results: Ten anti-VCAM1 nanobodies were generated, radiolabeled with technetium-99m, and screened in vitro on mouse and human recombinant VCAM1 proteins and endothelial cells and in vivo in apolipoprotein E-deficient (ApoE(-/-)) mice. Anontargeting control nanobody was used in all experiments to demonstrate specificity. All nanobodies displayed nanomolar affinities for murine VCAM1. Flow cytometry analyses using human human umbilical vein endothelial cells indicated murine and human VCAM1 cross-reactivity for 6 of 10 nanobodies. The lead compound cAbVCAM1-5 was cross-reactive for human VCAM1 and exhibited high lesion-to-control (4.95+/-0.85), lesion-to-heart (8.30+/-1.11), and lesion-to-blood ratios (4.32+/-0.48) (P