A Hypoxia- and α-Fetoprotein-Dependent Oncolytic Adenovirus Exhibits Specific Killing of Hepatocellular Carcinomas
A Hypoxia- and α-Fetoprotein-Dependent Oncolytic Adenovirus Exhibits Specific Killing of Hepatocellular Carcinomas
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DOI：
10.1158/1078-0432.ccr-10-0664
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发表时间：
2010-12-15
影响因子：
11.5
通讯作者：
Yun, Chae-Ok
中科院分区：
文献类型：
--
作者：
Kwon, Oh-Joon;Kim, Pyung-Hwan;Yun, Chae-Ok
目的：溶瘤腺病毒（Oncolytic adenoviruses，Ad）是一种新型的肿瘤基因治疗方法，其治疗效果优于非复制型Ad。我们以前已经表明，E1 B 19 kDa缺失的溶瘤腺病毒表现出强烈的细胞杀伤作用，但缺乏肿瘤选择性。为了实现肝癌限制性细胞毒性和增强Ad在肿瘤微环境中的复制，我们使用修饰的人甲胎蛋白（hAFP）启动子来控制Ad的复制和缺氧反应元件（HRE）。实验设计：我们构建了Ad-HRE 6/hAFP Delta 19和Ad-HRE 12/hAFP Delta 19，其在启动子上游掺入6或12个拷贝的HRE。荧光素酶法和荧光激活细胞分选法检测启动子活性和肝癌特异性。采用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四氮唑溴化法和细胞病变法检测Ad-HRE 6/hAFP Delta 19和Ad-HRE 12/hAFP Delta 19的AFP表达依赖性和缺氧依赖性体外细胞毒性。结果：Ad-HRE 12/hAFP Delta 19与Ad-hAFP Delta 19相比，具有更强的肿瘤选择性和细胞杀伤活性。Ad-HRE 12/hAFP Delta 19的杀肿瘤活性导致在皮下和原位模型中肿瘤生长的显著抑制。治疗后原发肿瘤的组织学检查证实了缺氧区域附近的病毒颗粒积聚。此外，Ad-HRE 12/hAFP Delta 19没有引起严重的炎症免疫反应和毒性后全身injection.Conclusions：这里提出的结果显示了将HRE纳入hAFP启动子驱动的溶瘤病毒的优势。该系统的独特之处在于它以组织特异性和肿瘤环境选择性的方式起作用。Ad-HRE 12/hAFP Delta 19的选择性和杀瘤活性大大增强，使其成为一种有希望的肝癌治疗剂。临床癌症研究; 16（24）; 6071-82。(C)2010年AACR。
Purpose: Oncolytic adenoviruses (Ad) constitute a new promising modality of cancer gene therapy that displays improved efficacy over nonreplicating Ads. We have previously shown that an E1B 19-kDa-deleted oncolytic Ad exhibits a strong cell-killing effect but lacks tumor selectivity. To achieve hepatoma-restricted cytotoxicity and enhance replication of Ad within the context of tumor microenvironment, we used a modified human alpha-fetoprotein (hAFP) promoter to control the replication of Ad with a hypoxia response element (HRE).Experimental Design: We constructed Ad-HRE6/hAFP Delta 19 and Ad-HRE12/hAFP Delta 19 that incorporated either 6 or 12 copies of HRE upstream of promoter. The promoter activity and specificity to hepatoma were examined by luciferase assay and fluorescence-activated cell sorting analysis. In addition, the AFP expression-and hypoxia-dependent in vitro cytotoxicity of Ad-HRE6/hAFP Delta 19 and Ad-HRE12/hAFP Delta 19 was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cytopathic effect assay. In vivo tumoricidal activity on subcutaneous and liver orthotopic model was monitored by noninvasive molecular imaging.Results: Ad-HRE12/hAFP Delta 19 exhibited enhanced tumor selectivity and cell-killing activity when compared with Ad-hAFP Delta 19. The tumoricidal activity of Ad-HRE12/hAFP Delta 19 resulted in significant inhibition of tumor growth in both subcutaneous and orthotopic models. Histologic examination of the primary tumor after treatment confirmed accumulation of viral particles near hypoxic areas. Furthermore, Ad-HRE12/hAFP Delta 19 did not cause severe inflammatory immune response and toxicity after systemic injection.Conclusions: The results presented here show the advantages of incorporating HREs into a hAFP promoter-driven oncolytic virus. This system is unique in that it acts in both a tissue-specific and tumor environment-selective manner. The greatly enhanced selectivity and tumoricidal activity of Ad-HRE12/hAFP Delta 19 make it a promising therapeutic agent in the treatment of liver cancers. Clin Cancer Res; 16(24); 6071-82. (C) 2010 AACR.