Translational control by an oncolytic adenovirus

溶瘤腺病毒的翻译控制

基本信息

批准号：
6938568
负责人：
MICHAEL A THOMAS
金额：
$ 4.1万
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/6938568
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，很少有选择性的，甚至很少表现出最小的毒性。这强调了对满足这两个要求的新型有效治疗剂的需求。溶瘤病毒似乎就是这样的一种治疗方法。 55K 突变腺病毒正处于癌症治疗的 III 期临床试验中。该实验室表明，55K 突变病毒可以选择性地在 S 期感染细胞中复制并杀死它们。然而，这种选择性的基础尚不清楚。了解 55K 突变病毒 S 期选择性的基础可能会指出如何操纵该病毒以更有效地杀死肿瘤细胞。该项目的首要目标是确定 S 期导致 55K 突变病毒晚期病毒基因翻译增加的机制。将产生具有适当前导序列的晚期病毒信息，并测量蛋白质产生率，以检验 G1 感染细胞中 55K 突变病毒的晚期病毒基因翻译减少是由于核糖体分流减少所致的假设。值得注意的是，从 55K 突变病毒中删除 E4orf1 和 E4orf2 基因可将病毒蛋白产量恢复到接近野生型病毒水平，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所致。这项工作测试了以下假设：55K 突变病毒的 S 期选择性是由于晚期病毒信息的翻译增加所致，而晚期病毒信息的翻译在 G1 期受到 E4orf1 的限制。