Chemotrap-1: an engineered soluble receptor that blocks chemokine-induced migration of metastatic cancer cells in vivo.
Chemotrap-1: an engineered soluble receptor that blocks chemokine-induced migration of metastatic cancer cells in vivo.
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DOI：
10.1158/0008-5472.can-10-0175
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发表时间：
2010-10-15
期刊：
影响因子：
11.2
通讯作者：
Bates DO
中科院分区：
文献类型：
--
作者：
Lanati S;Dunn DB;Roussigné M;Emmett MS;Carriere V;Jullien D;Budge J;Fryer J;Erard M;Cailler F;Girard JP;Bates DO
癌症和树突状细胞识别并向淋巴分泌的趋化因子迁移，并利用这一机制侵入淋巴系统，癌细胞通过淋巴系统转移。淋巴分泌的趋化因子配体CCL21已被确定为黑色素瘤和乳腺癌细胞转移表型转换的关键调控分子。然而，目前尚不清楚CCL21抑制是否是抑制转移的潜在治疗策略。在这里，我们描述了一种工程CCL21可溶性抑制剂，Chemotrap-1，它可以抑制转移性黑色素瘤细胞在体内的迁移。双杂交、拉下和共免疫沉淀试验使我们能够鉴定出具有CCL21趋化因子结合特性的自然存在的人锌指蛋白。进一步分析发现了一个短肽（约70个氨基酸），具有预测的卷曲卷曲结构，足以与CCL21关联。然后将这种CCL21趋化因子结合肽融合到人IgG1的Fc区域，生成Chemotrap-1，这是一种人趋化因子结合Fc融合蛋白。表面等离子体共振（SPR）和趋化实验表明，Chemotrap-1与CCL21结合，并以亚nM的亲和力抑制CCL21诱导的黑色素瘤细胞的体外迁移。此外，在体外和体内实验中，Chemotrap-1阻断黑色素瘤细胞向淋巴内皮细胞的迁移。最后，Chemotrap-1在体内显著减少表达CCR7的黑色素瘤细胞的淋巴侵袭、追踪和转移。综上所述，这些结果表明，通过Chemotrap-1抑制CCL21趋化因子是一种潜在的转移治疗策略，并进一步支持淋巴介导的转移是一个趋化因子依赖过程的假设。
Cancer and dendritic cells recognise and migrate towards chemokines secreted from lymphatics, and use this mechanism to invade the lymphatic system, and cancer cells, metastasise through it. The lymphatic secreted chemokine ligand CCL21 has been identified as a key regulatory molecule in the switch to a metastatic phenotype in melanoma and breast cancer cells. However, it is not known whether CCL21 inhibition is a potential therapeutic strategy for inhibition of metastasis. Here, we describe an engineered CCL21 soluble inhibitor, Chemotrap-1, which inhibits migration of metastatic melanoma cells in vivo. Two-hybrid, pull down and co-immunoprecipitation assays allowed us to identify a naturally occurring human zinc finger protein with CCL21 chemokine-binding properties. Further analyses revealed a short peptide (~ 70 amino-acids), with a predicted coiled-coil structure, which is sufficient for association with CCL21. This CCL21 chemokine-binding peptide was then fused to the Fc region of human IgG1 to generate Chemotrap-1, a human chemokine-binding Fc-fusion protein. Surface plasmon resonance (SPR) and chemotaxis assays showed that Chemotrap-1 binds CCL21 and inhibits CCL21-induced migration of melanoma cells in vitro with sub nM affinity. In addition, Chemotrap-1 blocked migration of melanoma cells towards lymphatic endothelial cells in vitro and in vivo. Finally, Chemotrap-1 strongly reduced lymphatic invasion, tracking and metastasis of CCR7 expressing melanoma cells in vivo. Together, these results show that CCL21 chemokine inhibition by Chemotrap-1 is a potential therapeutic strategy for metastasis, and provide further support for the hypothesis that lymphatic mediated metastasis is a chemokine-dependent process.