Target cell-restricted and -enhanced apoptosis induction by a scFv:sTRAIL fusion protein with specificity for the pancarcinoma-associated antigen EGP2
Target cell-restricted and -enhanced apoptosis induction by a scFv:sTRAIL fusion protein with specificity for the pancarcinoma-associated antigen EGP2
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DOI：
10.1002/ijc.11702
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发表时间：
2004-03-20
影响因子：
6.4
通讯作者：
Helfrich, W
中科院分区：
文献类型：
--
作者：
Bremer, E;Kuulen, J;Helfrich, W
重组可溶性肿瘤坏死因子相关凋亡诱导配体（TRAIL）具有明显的肿瘤选择性凋亡诱导活性，已引起临床应用的极大兴趣。然而，为了充分利用其治疗潜力，应考虑TRAIL受体系统和可溶性TRAIL（sTRAIL）两者的特征：首先，各种TRAIL受体在整个人体中的广泛表达;第二，激动性受体TRAIL-R1和TRAIL-R2的不同结合亲和力和交联要求;以及第三，特定sTRAIL制剂的溶液行为。因此，我们构建了一种新的TRAIL融合蛋白，命名为scFvC 54：sTRAIL，其包含与人sTRAIL的N-末端遗传连接的人scFv抗体片段C54。scFvCS 4：sTRAIL融合蛋白被设计为仅在scFvCS 4：sTRAIL与大量表达的癌相关细胞表面抗原EGP 2（别名EpCAM）特异性结合后通过激动性TRAIL受体的交联来诱导细胞凋亡。靶抗原限制性细胞凋亡诱导被证明为各种EGP 2阳性肿瘤细胞，并可被EGP 2竞争抗体抑制。靶抗原结合将可溶性scFvCS 4：sTRAIL转化为膜结合形式的TRAIL，其不仅能够通过TRAIL-R1而且还能够通过TRAIL-R2发出凋亡信号。尺寸排阻快速蛋白液相色谱法（FPLC）表明，scFvCS 4：sTRAIL作为稳定和均匀的三聚体产生的可检测的TRAIL聚集体的情况下。scFvCS 4：sTRAIL融合蛋白的有利特性潜在地减少了抗肿瘤活性所需的sTRAIL的量，并且可能对于治疗各种人类癌症具有价值。(C)2003 Wiley-Liss，Inc.
The apparent tumor selective apoptosis-inducing activity of recombinant soluble TNF-related apoptosis-inducing ligand (TRAIL) has aroused much interest for use in clinical application. However, to exploit fully its therapeutic potential, the characteristics of both the TRAIL receptor system and soluble TRAIL (sTRAIL) should be taken into account: first, the widespread expression of the various TRAIL receptors throughout the human body; second, the differential binding affinities and crosslinking requirements of the agonistic receptors TRAIL-RI and TRAIL-R2; and third, the solution behavior of particular sTRAIL preparations. Therefore, we constructed a novel TRAIL fusion protein, designated scFvC54:sTRAIL, comprising the human scFv antibody fragment C54 genetically linked to the N-terminus of human sTRAIL. The scFvCS4:sTRAIL fusion protein was designed to induce apoptosis by crosslinking of agonistic TRAIL receptors only after specific binding of scFvCS4:sTRAIL to the abundantly expressed carcinoma-associated cell surface antigen EGP2 (alias EpCAM). Target antigen-restricted apoptosis induction was demonstrated for various EGP2-positive tumor cells and could be inhibited by an EGP2 competing antibody. Target antigen binding converted soluble scFvCS4: sTRAIL into a membrane-bound form of TRAIL that was capable of signaling apoptosis not only through TRAIL-R1 but also through TRAIL-R2. Size-exclusion fast protein liquid chromatography (FPLC) indicated that scFvCS4:sTRAIL was produced as stable and homogeneous trimers in the absence of detectable TRAIL aggregates. The favorable characteristics of the scFvCS4:sTRAIL fusion protein potentially reduce the amount of sTRAIL required for antitumor activity and may be of value for the treatment of various human carcinomas. (C) 2003 Wiley-Liss, Inc.