EpCAM- and EGFR-targeted selective gene therapy for biliary cancers using Z33-fiber-modified adenovirus
EpCAM- and EGFR-targeted selective gene therapy for biliary cancers using Z33-fiber-modified adenovirus
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DOI：
10.1002/ijc.25758
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发表时间：
2011-09-01
影响因子：
6.4
通讯作者：
Yokoyama, Kazunari K.
中科院分区：
文献类型：
--
作者：
Kawashima, Rei;Abei, Masato;Yokoyama, Kazunari K.
基于腺病毒（Ad）的癌症基因治疗的一个关键问题是提高向癌细胞传递基因的特异性，以获得更好的疗效和安全性。我们探索了重新定位 Ad 载体的方法，用于人类胆道癌的选择性基因治疗，使用包含葡萄球菌蛋白 A (Ad-FZ33) 的 IgG Fc 结合基序 (Z33) 的 Ad 与肿瘤特异性抗体相结合。流式细胞术分析显示人胆管癌细胞上皮细胞粘附分子（EpCAM）和表皮生长因子受体（EGFR）的高表达水平。与使用对照抗体或不使用抗体的治疗相比，表达 LacZ 的 Ad-FZ33 与针对 EpCAM 或 EGFR 的抗体结合，然后进行 β-gal 测定，在这些胆管癌细胞中证明了高效的基因转导。 Ad-FZ33表达尿嘧啶磷酸核糖转移酶（UPRT），这种酶大大增强了5-氟尿嘧啶（FU）的毒性，与EpCAM或EGFR抗体结合，显着增强胆道癌细胞对5-FU的敏感性。相比之下，治疗不影响不表达EpCAM或EGFR的细胞（包括正常肝细胞）的5-FU敏感性。最后，用抗 EpCAM 或 EGFR 抗体治疗表达 UPRT 的 Ad-FZ33，然后施用 5-FU，显着抑制了裸鼠体内胆管癌异种移植物的生长。这些结果表明，由 Z33 纤维修饰的 Ad 与抗 EpCAM 或抗 EGFR 抗体介导的基因治疗提供了针对胆管癌的潜在有效治疗方式。
A critical issue in adenovirus (Ad)-based cancer gene therapy is to improve the specificity of gene delivery to cancer cells for better efficacy and safety. We explored methods of retargeting Ad vectors for selective gene therapy of human biliary cancers using the Ad incorporating an IgG Fc-binding motif (Z33) from the Staphylococcus protein A (Ad-FZ33) combined with tumor-specific antibodies. Flow cytometry analysis revealed high-expression levels of epithelial cell adhesion molecule (EpCAM) and epidermal growth factor receptor (EGFR) on human biliary cancer cells. Ad-FZ33 expressing LacZ combined with antibodies against EpCAM or EGFR, followed by beta-gal assay, demonstrated highly efficient gene transduction in these biliary cancer cells, compared to the treatment with control antibody or without antibody. Ad-FZ33 expressing uracil phosphoribosyl transferase (UPRT), an enzyme which greatly enhances the toxicity of 5-fluorouracil (FU), combined with antibodies against EpCAM or EGFR, remarkably enhanced the sensitivity of biliary cancer cells to 5-FU. By contrast, the treatment did not affect the 5-FU sensitivity of the cells not expressing EpCAM or EGFR including normal hepatocytes. Finally, treatments with the UPRT-expressing Ad-FZ33 with antibodies against EpCAM or EGFR, followed by 5-FU administration, significantly suppressed the growth of biliary cancer xenografts in nude mice. These results indicate that the gene therapy mediated by the Z33 fiber modified Ad with anti-EpCAM or anti-EGFR antibodies offers a potentially effective therapeutic modality against biliary cancers.