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Development of the PKR-dependent replicating adenovirus

PKR依赖性复制腺病毒的开发

基本信息

批准号：
18591433
负责人：
SASAKI Katsunori
金额：
$ 2.49万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591433/
关键词：
Adenovirus Vector Gene Therapy Pancreatic Cancer PKR ras VAI Interferon IFN

项目摘要

The design of double-stranded RNA-activated protein kinase (PKR)-depended replicating adenovirus is described as a new approach to cancer gene therapy. The virus-associated type I (VAI) RNA cannot activate PKR, although it binds to the protein and thereby prevents its activation by authentic dsRNA. In this study, we engineered the conditionally replicative adenovirus (CRAD) with the modification of VAI gene and investigated the utility of CRAD targeted to pancreatic cancer. We constructed a modificated VAI gene (mVAI) which the central domain was deleted. This mVAI was replaced its own endogenous VAI gene harboring in adenovirus type 5 genome (Adv-mVAI). When the cytolytic activity of Adv-mVAI was evaluated, it failed to inhibit the growth of the pancreatic cancer cell; BxPC-3 with wild K-ras. Interestingly, we demonstrated that the transduction of active-formed ras into BxPC-3 cells by the infection of retroviruses expressing H-ras/V12 was sufficient to induce the cytolytic activity of Adv-mVAI. In addition, the cytolytic activity of Adv-mVAI was observed in the pancreatic cancer cell; AsPC-1 with mutant K-ras. However, this cytolytic activity of Adv-mVAI was reduced after interferon treatment.These data indicate that the activation of ras and interferon will be useful for controlling the anti-tumor effect of adenovirus harboring the variant VAI gene.

双链RNA激活蛋白激酶（PKR）依赖的复制型腺病毒的设计为肿瘤基因治疗开辟了新的途径。病毒相关的I型（VAI）RNA不能激活PKR，尽管它与蛋白质结合，从而阻止其被真正的dsRNA激活。在本研究中，我们设计了VAI基因修饰的条件复制型腺病毒（CRAD），并研究了CRAD对胰腺癌的靶向作用。我们构建了一个中心结构域缺失的VAI基因（mVAI）。该mVAI被5型腺病毒基因组中的内源性VAI基因所取代（Adv-mVAI）。当评价Adv-mVAI的细胞溶解活性时，其不能抑制胰腺癌细胞的生长;具有野生K-ras的BxPC-3。有趣的是，我们证明了通过感染表达H-ras/V12的逆转录病毒将活性形成的ras转导到BxPC-3细胞中足以诱导Adv-mVAI的细胞溶解活性。此外，在胰腺癌细胞中观察到Adv-mVAI的细胞溶解活性;具有突变K-ras的AsPC-1。这些数据表明，ras和干扰素的激活将有助于控制携带变异VAI基因的腺病毒的抗肿瘤作用。