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激活的蛋白激酶（PKR） - 抑制复制腺病毒的设计被描述为一种癌症基因治疗的新方法。与病毒相关的I型RNA（VAI）RNA无法激活PKR，尽管它与蛋白质结合，从而防止其通过正宗DSRNA的激活。在这项研究中，我们对有条件复制的腺病毒（CRAD）进行了设计，并通过VAI基因的修饰进行了研究，并研究了针对胰腺癌的CRAD的实用性。我们构建了一个经过修改的VAI基因（MVAI），该基因被删除了。该MVAI被替换为在腺病毒5型基因组（ADV-MVAI）中携带的内源性VAI基因。当评估ADV-MVAI的细胞溶解活性时，它未能抑制胰腺癌细胞的生长。 BXPC-3与野生K-Ras。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了ADV-MVAI的细胞溶解活性。 ASPC-1与突变k-ras。然而，干扰素治疗后降低了ADV-MVAI的细胞溶解活性。这些数据表明，Ras和Interferon的激活将有助于控制具有变体VAI基因的腺病毒的抗肿瘤效应。