Selective function-blocking monoclonal human antibody highlights the important role of membrane type-1 matrix metalloproteinase (MT1-MMP) in metastasis.
Selective function-blocking monoclonal human antibody highlights the important role of membrane type-1 matrix metalloproteinase (MT1-MMP) in metastasis.
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DOI：
10.18632/oncotarget.13157
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发表时间：
2017-01-10
期刊：
影响因子：
--
通讯作者：
Strongin AY
中科院分区：
文献类型：
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作者：
Remacle AG;Cieplak P;Nam DH;Shiryaev SA;Ge X;Strongin AY
促进侵袭的 MT1-MMP 是一种细胞表面相关的胶原酶，具有多种关键的细胞功能。人们一致认为，MT1-MMP 是迁移癌细胞中异常细胞周蛋白水解的关键蛋白酶，因此是一个有前途的药物靶点。由于 MMP 家族的高度同源性以及选择性小分子抑制剂设计的成功有限，很明显，选择性和成功的 MT1-MMP 疗法需要抑制剂特异性。使用表现出延伸的 23-27 个残基长的 VH CDR-H3 片段的人 Fab 抗体库（超过 1.25×109 个个体变体），我们分离了一组抑制性抗体片段，其中 3A2 Fab 作为一种特异性、有效、低纳摩尔范围的 MT1-MMP 抑制剂，优于其他抗体片段。在这里，我们报告了 3A2 抗体的深入表征。我们的多项体外和基于细胞的测试和测定，以及抗体/蛋白酶相互作用的广泛结构建模表明，抗体表位涉及靠近蛋白酶催化位点的残基，并且与 MMP 的组织抑制剂 2 (TIMP-2) 相比，3A2 Fab 通过与活性位点腔外的催化结构域结合来灭活蛋白酶功能。与其他人对转移的研究一致，我们对急性肺黑色素瘤转移的动物研究支持 MT1-MMP 在转移过程中的关键作用。相反，选择性抗 MT1-MMP 单一疗法显着减轻了黑色素瘤的转移负担。 3A2 Fab 的进一步亲和力成熟可能会产生先导抑制剂，并证明 MT1-MMP 靶向转移性癌症的概念。
The invasion-promoting MT1-MMP is a cell surface-associated collagenase with a plethora of critical cellular functions. There is a consensus that MT1-MMP is a key protease in aberrant pericellular proteolysis in migrating cancer cells and, accordingly, a promising drug target. Because of high homology in the MMP family and a limited success in the design of selective small-molecule inhibitors, it became evident that the inhibitor specificity is required for selective and successful MT1-MMP therapies. Using the human Fab antibody library (over 1.25×109 individual variants) that exhibited the extended, 23-27 residue long, VH CDR-H3 segments, we isolated a panel of the inhibitory antibody fragments, from which the 3A2 Fab outperformed others as a specific and potent, low nanomolar range, inhibitor of MT1-MMP. Here, we report the in-depth characterization of the 3A2 antibody. Our multiple in vitro and cell-based tests and assays, and extensive structural modeling of the antibody/protease interactions suggest that the antibody epitope involves the residues proximal to the protease catalytic site and that, in contrast with tissue inhibitor-2 of MMPs (TIMP-2), the 3A2 Fab inactivates the protease functionality by binding to the catalytic domain outside the active site cavity. In agreement with the studies in metastasis by others, our animal studies in acute pulmonary melanoma metastasis support a key role of MT1-MMP in metastatic process. Conversely, the selective anti-MT1-MMP monotherapy significantly alleviated melanoma metastatic burden. It is likely that further affinity maturation of the 3A2 Fab will result in the lead inhibitor and a proof-of-concept for MT1-MMP targeting in metastatic cancers.