Potent killing of HBV-related hepatocellular carcinoma by a chimeric protein of anti-HBsAg single-chain antibody and truncated Bid
Potent killing of HBV-related hepatocellular carcinoma by a chimeric protein of anti-HBsAg single-chain antibody and truncated Bid
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抗HBsAg单链抗体与截短Bid的嵌合蛋白有效杀灭HBV相关肝细胞癌
DOI：
10.1016/j.biomaterials.2013.03.046
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发表时间：
2013-07-01
期刊：
影响因子：
14
通讯作者：
Yang, Angang
中科院分区：
文献类型：
--
作者：
Yan, Bo;Ouyang, Qing;Yang, Angang
B型肝炎病毒（HBV）介导的肝细胞癌（HCC）表现出HBV表面抗原（HBsAg）的过度表达，需要靶向治疗。我们先前开发了抗HBsAg的人单链抗体scFv 15。在这里，我们测试了scFv 15介导的凋亡效应物递送用于以HBsAg为靶向的HCC治疗的战略可行性，以及流感血凝素的HA 2基序的应用以增强内体逃逸和抗肿瘤作用。通过将来自白喉毒素（Fdt）的弗林蛋白酶可切割基序（furin-cleavable motif）、HA 2和截短的凋亡蛋白Bid（tBid）依次融合，产生了一类以HBsAg为靶向的免疫促凋亡分子。结果表明，在原核细胞中表达的scFv 15-Fdt-HA 2-tBid具有较好的抗肿瘤活性，并与scFv 15-Fdt-tBid进行了比较。scFv 15-Fdt-HA 2-tBid和scFv 15-Fdt-tBid都保留了对HBsAg的亲和力和特异性，并通过凋亡结合和选择性杀死HBsAg阳性的HCC细胞。值得注意的是，scFv 15-Fdt-HA 2-tBid在HBsAg阳性PLC/PRF/5细胞中的IC 50比scFv 15-Fdt-tBid低10倍。抗肿瘤活性的体内成像显示scFv 15-Fdt-HA 2-tBid对原位HCC的生长抑制率为95%，而scFv 15-Fdt-tBid的抑制率为75%。这项研究代表了免疫促凋亡策略在治疗HBeAg阳性HCC中的扩展应用，并显示了HA 2作为内体包封的抗体缀合物的功能增强剂的显著潜力。(C)2013爱思唯尔有限公司保留所有权利。
Targeted therapy is needed for hepatitis B virus (HBV)-mediated hepatocellular carcinoma (HCC) which shows overexpression of HBV surface antigen (HBsAg). We previously developed scFv15, a human single-chain antibody against HBsAg. Here we tested the strategic feasibility of scFv15-mediated delivery of apoptotic effectors for HBsAg-targeted HCC therapy and application of HA2 motif of influenza hemagglutinin to enhance endosome escape and antitumor effect. A class of HBsAg-targeted immunoproapoptotic molecule was generated by sequentially fusing scFv15, the furin-cleavable motif from diphtheria toxin (Fdt), HA2 and a truncated apoptotic protein Bid (tBid). The resulting scFv15-Fdt-HA2-tBid was prokaryotically expressed and functionally characterized for HBsAg-binding capacity, endosome escape activity and antitumor effect as compared with scFv15-Fdt-tBid. Both scFv15-Fdt-HA2-tBid and scFv15-Fdt-tBid retained affinity and specificity for HBsAg, and bound and selectively killed HBsAg-positive HCC cells via apoptosis. Notably, the IC50 of scFv15-Fdt-HA2-tBid in HBsAg-positive PLC/PRF/5 cells was 10 times lower than that of scFv15-Fdt-tBid. In vivo imaging of antitumor activity demonstrated 95% growth inhibition of orthotopic HCC by scFv15-Fdt-HA2-tBid compared with 75% suppression by scFv15-Fdt-tBid. This study represents an extended application of the immunoproapoptotic strategy in the treatment of HBsAg-positive HCC and shows significant potential of HA2 as a functional enhancer for endosome-encapsulated antibody-conjugates. (C) 2013 Elsevier Ltd. All rights reserved.