Targeting the tumor vasculature with engineered cystine-knot miniproteins
Targeting the tumor vasculature with engineered cystine-knot miniproteins
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DOI：
10.1038/s41467-019-13948-y
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发表时间：
2020-01-15
影响因子：
16.6
通讯作者：
Sahin, Ugur
中科院分区：
文献类型：
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作者：
Lui, Bonny Gaby;Salomon, Nadja;Sahin, Ugur
人纤连蛋白的额外结构域B剪接变体（EDB）在肿瘤血管系统中选择性表达，是癌症成像和治疗的有吸引力的靶点。在这里，我们描述了EDB特异性光学成像探针的产生和表征。通过噬菌体展示技术筛选组合胱氨酸结微蛋白文库，我们发现了具有独特基序的精美的EDB特异性配体。具有皮摩尔范围内的结合常数的探针通过所选配体的化学寡聚化和荧光团缀合产生。我们通过荧光成像显示，探针对源自小鼠中的人U-87 MG胶质母细胞瘤异种移植物的组织切片中的EDB染色。此外，我们通过体内和离体荧光成像证明了静脉给药的探针在具有U-87 MG胶质母细胞瘤异种移植物的小鼠的肿瘤组织中的选择性积累和保留。这些数据保证了进一步追求所选择的胱氨酸结微蛋白用于体内成像应用。
The extra domain B splice variant (EDB) of human fibronectin selectively expressed in the tumor vasculature is an attractive target for cancer imaging and therapy. Here, we describe the generation and characterization of EDB-specific optical imaging probes. By screening combinatorial cystine-knot miniprotein libraries with phage display technology we discover exquisitely EDB-specific ligands that share a distinctive motif. Probes with a binding constant in the picomolar range are generated by chemical oligomerization of selected ligands and fluorophore conjugation. We show by fluorescence imaging that the probes stain EDB in tissue sections derived from human U-87 MG glioblastoma xenografts in mice. Moreover, we demonstrate selective accumulation and retention of intravenously administered probes in the tumor tissue of mice with U-87 MG glioblastoma xenografts by in vivo and ex vivo fluorescence imaging. These data warrants further pursuit of the selected cystine-knot miniproteins for in vivo imaging applications.