Translational control by an oncolytic adenovirus

溶瘤腺病毒的翻译控制

基本信息

批准号：
7111710
负责人：
MICHAEL A THOMAS
金额：
$ 2.54万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-15 至 2007-08-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7111710
关键词：
Adenoviridae DNA replication HeLa cells RNase protection assay SDS polyacrylamide gel electrophoresis autoradiography cell cycle cell growth regulation gene deletion mutation genetic translation immunoprecipitation polymerase chain reaction predoctoral investigator protein biosynthesis virus protein virus replication

项目摘要

DESCRIPTION (provided by applicant): Of the primary modalities used to treat cancer, few are selective and even less exhibit minimal toxicity. This underscores the need for new and effective therapeutic agents that meets both requirements. Oncolytic viruses appear to be such a treatment. The 55K-mutant adenovirus is in phase III clinical trials for use in cancer therapy. This lab has shown that the 55K-mutant virus selectively replicates in and kill S-phase infected cells. However, the basis for this selectivity is not known. Understanding the basis for the S-phase selectivity of the 55K-mutant virus may point to how the virus can be manipulated to be more effective at killing tumor cells. The first aim of this project is to determine the mechanism in S-phase that leads to an increase in late viral gene translation of the 55K-mutant virus. Late viral messages with appropriate leader sequences will be made and the rate of protein production measured to test the hypothesis that the decrease in late viral gene translation of the 55K-mutant virus in G1 infected cells is due to a decrease in ribosomal shunting. Remarkably, deletion of the E4orf1 and E4orf2 genes from the 55K-mutant virus restores viral protein production to near wild-type virus levels and abrogates the G1-restriction. Using recombinant viruses, the second aim of this project will determine if the G1-restriction is due to the ability of E4orf1 or E4orf2 to attenuate late viral gene translation. This work tests the hypothesis that the S-phase selectivity of the 55K-mutant virus is due to an increase in translation of late viral messages which is restricted in G1 by E4orf1.

描述（由申请人提供）：在用于治疗癌症的主要方式中，很少有选择性，甚至更少表现出最小的毒性。这强调了满足这两个要求的新的有效治疗剂的需求。溶瘤病毒似乎是一种治疗方法。 55K突变的腺病毒正在III期临床试验中用于癌症治疗。该实验室表明，55K突变病毒在中有选择地复制并杀死S期感染的细胞。但是，这种选择性的基础尚不清楚。了解55K突变病毒的S相选择性的基础可能指出如何操纵该病毒在杀死肿瘤细胞方面更有效。该项目的第一个目的是确定S期的机制，从而导致55K突变病毒的晚期病毒基因翻译增加。将制定具有适当铅序列的晚期病毒消息，并测量的蛋白质产生速率是检验以下假设：G1感染细胞中55K突变病毒的晚期病毒基因翻译的减少是由于核糖体浮肿的降低所致。值得注意的是，从55K突变病毒中的E4ORF1和E4ORF2基因的缺失将病毒蛋白的产生恢复为近乎野生型病毒水平并消除G1限制。使用重组病毒，该项目的第二个目标将确定G1限制是否是由于E4ORF1或E4ORF2减弱晚期病毒基因翻译的能力。这项工作检验了以下假设：55K突变病毒的S相选择性是由于晚期病毒消息的翻译增加所致，而病毒式消息的翻译增加了E4ORF1在G1中受到限制。