Construction and characterization of a truncated tissue factor-coagulation-based composite system for selective thrombosis in tumor blood vessels
Construction and characterization of a truncated tissue factor-coagulation-based composite system for selective thrombosis in tumor blood vessels
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用于肿瘤血管选择性血栓形成的截短组织因子-凝血复合系统的构建和表征
DOI：
10.3892/ijo.2019.4855
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发表时间：
2019-08
影响因子：
5.2
通讯作者：
Yan Jianghua
中科院分区：
文献类型：
--
作者：
Xu Peilan;Zou Mingyuan;Wang Shengyu;Li Tingting;Liu Cong;Wang Li;Wang Lanlan;Luo Fanghong;Wu Ting;Yan Jianghua
通过靶配体传递截断的组织因子(TTF)选择性地诱导肿瘤血管血栓形成是一种很有前途的新的抗肿瘤策略。本研究建立了抗神经粘连蛋白-1(NRP-1)-链霉亲和素(SA)-生物素(B)复合体系。在该系统中，抗NRP-1单抗将TTF定位于肿瘤血管内皮细胞表面，诱导血管栓塞。由于SA和B具有很高的结合亲和力，因此它们被用来增强血栓形成活性。以水溶性1-乙基-3-(3-二甲氨基丙基)碳二亚胺和N-羟基磺基琥珀酰亚胺为偶联剂，将单抗与SA偶联。生物素化TTF(TTF-B)是在融合蛋白TTF产生和纯化后用B标记试剂盒制备的。共聚焦显微镜和流式细胞仪检测表明，抗NRP-1-mAb-SA结合物具有mAb靶向活性。用竞争酶联免疫吸附试验证实了B-偶联结合能力的保留，因子X激活分析表明TTF-B保留了TTF的促凝血活性。活体成像评估mAb-SA的分布和肿瘤靶向能力，这产生了令人振奋的结果。在小鼠皮下移植的体内研究结果表明，该复合系统显著诱导肿瘤血管血栓形成并抑制肿瘤生长，而在正常器官中未观察到这些组织学变化。
The selective induction of tumor vascular thrombosis using truncated tissue factor (tTF) delivered via a target ligand is a promising novel antitumor strategy. In the present study, an anti-neuropilin-1 (NRP-1) monoclonal antibody (mAb)-streptavidin (SA):tTF-biotin (B) composite system was established. In this system, anti-NRP-1-mAb located tTF to the tumor vascular endothelial cell surface and induced vascular embolization. Due to their high binding affinity, SA and B were used to enhance thrombogenic activity. mAb was conjugated with SA using a coupling method with water-soluble 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide. Biotinylated tTF (tTF-B) was prepared using a B-labeling kit subsequent to the generation and purification of fusion protein tTF. Confocal microscopy and flow cytometry indicated that the anti-NRP-1-mAb-SA conjugate retained mAb targeting activity. The preservation of B-conjugate binding capacity was confirmed using a competitive ELISA, and factor X-activation analysis revealed that tTF-B retained the procoagulant activity exhibited by tTF. Live imaging was performed to assess mAb-SA distribution and tumor-targeting capability, and this yielded promising results. The results of in vivo studies in mice with subcutaneous xenografts demonstrated that this composite system significantly induced tumor vascular thrombosis and inhibited tumor growth, whereas these histological changes were not observed in normal organs.
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影响因子：
3.1
作者：
Meili Zhang;H. Sakahara;Z. Yao;T. Saga;Y. Nakamoto;N. Sato;H. Nakada;I. Yamashina;J. Konishi
通讯作者：
Meili Zhang;H. Sakahara;Z. Yao;T. Saga;Y. Nakamoto;N. Sato;H. Nakada;I. Yamashina;J. Konishi
影响因子：
4.8
作者：
Wang, L;Zeng, HY;Mukhopadhyay, D
通讯作者：
Mukhopadhyay, D
影响因子：
20.3
作者：
Bieker, Ralf;Kessler, Torsten;Berdel, Wolfgang E.
通讯作者：
Berdel, Wolfgang E.
影响因子：
25.7
作者：
Berge, Mathieu;Allanic, David;Merkulova-Rainon, Tatyana
通讯作者：
Merkulova-Rainon, Tatyana
靶向 Neuropilin-1 的单克隆抗体通过抑制 FAK/p130cas 信号通路来抑制 MCF7 乳腺癌细胞与纤连蛋白的粘附
影响因子：
2.3
作者：
Zhang, Yafei;Lv, Haiyan;Yan, Jianghua;Chen, Yuqiang
通讯作者：
Chen, Yuqiang