E1A, E1B Double-Restricted Adenovirus with RGD-Fiber Modification Exhibits Enhanced Oncolysis for CAR–Deficient Biliary Cancers
E1A, E1B Double-Restricted Adenovirus with RGD-Fiber Modification Exhibits Enhanced Oncolysis for CAR–Deficient Biliary Cancers
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DOI：
10.1158/1078-0432.ccr-06-2103
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发表时间：
2007-05
影响因子：
11.5
通讯作者：
Mariko Wakayama;M. Abei;Rei Kawashima;Emiko Seo;Kuniaki Fukuda;H. Ugai;T. Murata;N. Tanaka;I. Hyodo;H. Hamada;K. Yokoyama
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文献类型：
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作者：
Mariko Wakayama;M. Abei;Rei Kawashima;Emiko Seo;Kuniaki Fukuda;H. Ugai;T. Murata;N. Tanaka;I. Hyodo;H. Hamada;K. Yokoyama
目的：胆道系统恶性肿瘤是高度恶性的疾病，预后极差。我们之前已经引入了AxdAdB 3，一种E1 A，E1 B双限制性溶瘤腺病毒，其对大约一半的胆管癌细胞系显示出优异的溶瘤功效，并且对正常细胞具有增强的安全性。本研究的目的是评估能够实现整合素依赖性感染的RGD-纤维修饰（AxdAdB 3-F/RGD）是否可以改善AxdAdB 3对胆管癌的感染性和疗效。实验设计：在体外一组胆管癌细胞系中，将腺病毒受体、柯萨奇病毒腺病毒受体（CAR）和整合素（αvβ3和αvβ5）的表达与表达LacZ的复制缺陷型腺病毒与野生型纤维或RGD修饰的纤维的感染性水平进行比较。本文比较了一种新型的E1 A、E1 B双限制性复制选择性腺病毒AxdAdB 3-F/RGD及其亲本AxdAdB 3在胆管癌细胞和正常细胞中的病毒复制和体外细胞毒活性。在异种移植肿瘤模型中比较这些溶瘤病毒的体内抗肿瘤作用。结果如下：CAR的表达与腺病毒的感染性显著相关，而整合素αvβ5在几乎所有胆管癌细胞中大量表达。AxdAdB 3仅有效复制和裂解保留CAR表达的胆管癌细胞，而AxdAdB 3-F/RGD在CAR阳性和CAR阴性胆管癌细胞中均表现出有效的复制和强效的溶瘤作用。AxdAdB 3-F/RGD在人正常细胞中显示减弱的复制和很少的细胞病变（即，肝细胞，WI-38细胞）以及AxdAdB 3。此外，在具有s.c. CAR缺陷型人胆管癌的异种移植物，i.t. AxdAdB 3-F/RGD疗法引起肿瘤生长的显著抑制。结论：RGD纤维修饰策略增强了E1 A，E1 B双限制性溶瘤腺病毒对CAR缺陷型胆管癌的感染性，复制和溶瘤作用。此外，它保留了双限制性病毒对正常细胞的优良安全性的优点。这些结果表明，这种药物用于治疗胆道癌的潜在用途。
Purpose: Cancers of biliary system represent highly malignant diseases of dismal prognosis. We have previously introduced AxdAdB3, an E1A, E1B double-restricted oncolytic adenovirus, which showed excellent oncolytic efficacy for approximately half of the biliary cancer lines with an enhanced safety to normal cells. The purpose of this study was to evaluate whether RGD-fiber modification (AxdAdB3-F/RGD), which enables integrin-dependent infection, can improve the infectivity and efficacy of AxdAdB3 for biliary cancers. Experimental Design: Expressions of adenoviral receptors, coxsackievirus adenovirus receptor (CAR) and integrins (αvβ3 and αvβ5), were compared with the level of infectivity of LacZ-expressing replication-defective adenoviruses with wild-type fibers or RGD-modified fibers in a panel of biliary cancer cell lines in vitro. Viral replication and cytotoxicity in vitro of AxdAdB3-F/RGD, a novel E1A, E1B double-restricted replication-selective adenovirus with RGD-modified fibers, were compared with those of its parent virus, AxdAdB3, in various biliary cancer cells and in normal cells. In vivo antitumor effects of these oncolytic viruses were compared in a xenograft tumor model. Results: Expression of CAR significantly correlated with the adenovirus infectivity, whereas integrin αvβ5 was abundantly expressed in almost all biliary cancer cells. Whereas AxdAdB3 effectively replicated and lysed only the biliary cancer cells with a preserved expression of CAR, AxdAdB3-F/RGD exhibited efficient replication and potent oncolysis in both CAR-positive and CAR-negative biliary cancer cells. AxdAdB3-F/RGD showed attenuated replication and little cytopathy in human normal cells (i.e., hepatocytes, WI-38 cells) as well as AxdAdB3. Furthermore, in nude mice with s.c. xenografts of CAR-deficient human biliary cancer, i.t. AxdAdB3-F/RGD therapy caused a marked inhibition of tumor growth. Conclusions: The RGD-fiber modification strategy enhanced the infectivity, replication, and oncolytic effects of the E1A, E1B double-restricted oncolytic adenovirus for CAR-deficient biliary cancers. In addition, it preserved the merit of excellent safety of the double-restricted virus for normal cells. These results suggest a potential use of this agent for the treatment of biliary cancers.