Development and characterization of a potent immunoconjugate targeting the Fn14 receptor on solid tumor cells.
Development and characterization of a potent immunoconjugate targeting the Fn14 receptor on solid tumor cells.
复制标题
DOI：
10.1158/1535-7163.mct-11-0161
复制
发表时间：
2011-07
影响因子：
5.7
通讯作者：
Winkles JA
中科院分区：
文献类型：
--
作者：
Zhou H;Marks JW;Hittelman WN;Yagita H;Cheung LH;Rosenblum MG;Winkles JA
TNF样弱凋亡诱导因子（TWEAK）和成纤维细胞生长因子（FGF）诱导因子14（Fn 14）是TNF超家族配体-受体对，参与许多细胞过程，包括增殖、迁移、分化、炎症和血管生成。Fn 14受体在正常组织中以相对低的水平表达，但已知其在许多肿瘤类型中显著升高，包括脑肿瘤和乳腺肿瘤。因此，它似乎是一个很好的候选治疗干预。我们首先分析了Fn 14在人肿瘤细胞系中的表达。Fn 14在多种细胞系中表达，包括乳腺癌、脑癌、膀胱癌、皮肤癌、肺癌、卵巢癌、胰腺癌、结肠癌、前列腺癌和宫颈癌细胞系。然后，我们开发了一种免疫偶联物，其中含有与重组白树蛋白（rGel）偶联的高亲和力抗Fn 14单克隆抗体（项目4），重组白树蛋白是一种高细胞毒性的核糖体失活N-糖苷酶。发现项目-4和缀合物均以等同程度结合细胞。共聚焦显微镜分析表明，ITEM 4-rGel特异性和快速（2小时内）内化到Fn 14阳性T-24膀胱癌细胞，但不进入Fn 14缺陷小鼠胚胎成纤维细胞。针对22种不同肿瘤细胞系的细胞毒性研究表明，ITEM 4-rGel对Fn 14表达细胞具有高度细胞毒性，其效力是游离rGel的8- 8 × 104倍。发现ITEM 4-rGel通过高迁移率族蛋白1蛋白释放诱导细胞凋亡来杀死细胞。最后，ITEM 4-rGel免疫缀合物施用促进了携带T-24人膀胱癌细胞异种移植物的裸鼠中的长期肿瘤生长抑制。我们的数据支持使用抗体-药物缀合物方法来选择性靶向和抑制表达Fn 14的肿瘤的生长。
TNF-like weak inducer of apoptosis (TWEAK) and fibroblast growth factor (FGF)-inducible 14 (Fn14) are a TNF superfamily ligand–receptor pair involved in many cellular processes including proliferation, migration, differentiation, inflammation, and angiogenesis. The Fn14 receptor is expressed at relatively low levels in normal tissues, but it is known to be dramatically elevated in a number of tumor types, including brain and breast tumors. Thus, it seems to be an excellent candidate for therapeutic intervention. We first analyzed Fn14 expression in human tumor cell lines. Fn14 was expressed in a variety of lines including breast, brain, bladder, skin, lung, ovarian, pancreatic, colon, prostate, and cervical cancer cell lines. We then developed an immunoconjugate containing a high-affinity anti-Fn14 monoclonal antibody (ITEM-4) conjugated to recombinant gelonin (rGel), a highly cytotoxic ribosome-inactivating N-glycosidase. Both ITEM-4 and the conjugate were found to bind to cells to an equivalent extent. Confocal microscopic analysis showed that ITEM4-rGel specifically and rapidly (within 2 hours) internalized into Fn14-positive T-24 bladder cancer cells but not into Fn14-deficient mouse embryonic fibroblasts. Cytotoxicity studies against 22 different tumor cell lines showed that ITEM4-rGel was highly cytotoxic to Fn14-expressing cells and was 8- to 8 × 104-fold more potent than free rGel. ITEM4-rGel was found to kill cells by inducing apoptosis with high-mobility group box 1 protein release. Finally, ITEM4-rGel immunoconjugate administration promoted long-term tumor growth suppression in nude mice bearing T-24 human bladder cancer cell xenografts. Our data support the use of an antibody–drug conjugate approach to selectively target and inhibit the growth of Fn14-expressing tumors.