Truncating the i-leader open reading frame enhances release of human adenovirus type 5 in glioma cells
Truncating the i-leader open reading frame enhances release of human adenovirus type 5 in glioma cells
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DOI：
10.1186/1743-422x-8-162
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发表时间：
2011-04-11
期刊：
影响因子：
4.8
通讯作者：
Hoeben, Rob C.
中科院分区：
文献类型：
--
作者：
van den Hengel, Sanne K.;de Vrij, Jeroen;Hoeben, Rob C.
背景：目前治疗方法下胶质瘤患者的生存率很低。早期的临床试验与复制腺病毒证明了使用腺病毒作为溶瘤剂的可行性和安全性。由于病毒复制和传播效率低，抗肿瘤疗效中等。先前的研究表明，腺病毒i-前导区开放阅读框的截短增强了HAdV-5在几种肿瘤细胞系中的致细胞病变活性。在这里，我们报告的影响，在胶质瘤细胞系和原代高级别胶质瘤细胞culture.Results的致细胞病变活性的i-领导人突变截断的i-领导人开放阅读框架创建在一个分子克隆的复制能力野生型HAdV-5的定点诱变。我们分析了这种RL-07突变病毒的致细胞病变活性。细胞活力测定显示RL-07突变病毒对U251和SNB 19神经胶质瘤细胞系的致细胞病变活性增加。RL-07在U251单层上的空斑大小是同基因对照病毒的7倍。类似地，RL-07病毒的致细胞病变活性在六种原代高级胶质瘤细胞培养物中强烈增加。在神经胶质瘤细胞系中，发现RL-07病毒更早地释放到培养基中。这是不是由于增强病毒蛋白质的合成，是显而易见的等效E1 A，纤维和腺病毒死亡蛋白的量，也不是更高的病毒yield.Conclusion：在胶质母细胞瘤细胞中的复制腺病毒的致细胞病变活性的增加，通过截断的i-领导人开放阅读框架。这种突变可能有助于增强溶瘤腺病毒在胶质母细胞瘤治疗中的抗肿瘤细胞病变功效。
Background: The survival of glioma patients with the current treatments is poor. Early clinical trails with replicating adenoviruses demonstrated the feasibility and safety of the use of adenoviruses as oncolytic agents. Antitumor efficacy has been moderate due to inefficient virus replication and spread. Previous studies have shown that truncation of the adenovirus i-leader open reading frame enhanced cytopathic activity of HAdV-5 in several tumor cell lines. Here we report the effect of an i-leader mutation on the cytopathic activity in glioma cell lines and in primary high-grade glioma cell cultures.Results: A mutation truncating the i-leader open reading frame was created in a molecular clone of replication-competent wild-type HAdV-5 by site-directed mutagenesis. We analyzed the cytopathic activity of this RL-07 mutant virus. A cell-viability assay showed increased cytopathic activity of the RL-07 mutant virus on U251 and SNB19 glioma cell lines. The plaque sizes of RL-07 on U251 monolayers were seven times larger than those of isogenic control viruses. Similarly, the cytopathic activity of the RL-07 viruses was strongly increased in six primary high-grade glioma cell cultures. In glioma cell lines the RL-07 virus was found to be released earlier into the culture medium. This was not due to enhanced viral protein synthesis, as was evident from equivalent E1A, Fiber and Adenovirus Death Protein amounts, nor to higher virus yields.Conclusion: The cytopathic activity of replicating adenovirus in glioblastoma cells is increased by truncating the i-leader open reading frame. Such mutations may help enhancing the antitumor cytopathic efficacy of oncolytic adenoviruses in the treatment of glioblastoma.