Development of therapeutic approach for progressive multifocal leukoencephalopathy using siRNA against JCV Agnogene.

在这个项目中，我们尝试使用小干扰rna （sirna）来抑制JCV agnoprotein （Agno）的感染作为治疗策略。为了抑制感染细胞中的JCV感染，我们合成了针对JCV编码蛋白Agno的双链sirna，并将其引入感染的人胶质源性细胞中。我们观察到，在感染后用靶向JCV的sirna治疗中，Agno抑制JCV感染细胞中的病毒产生。在此基础上，我们申请了两项专利，并发表了论文（中国生物医学杂志78:7270,2004）。此外，我们用小鼠脑内接种JCV感染细胞建立了JCV感染模拟模型，并在体内将抗JCV Agno的siRNA应用于模型。siRNA处理后，脑内jcvv感染细胞数量明显减少。此外，我们发现临床药物通过修饰体外一种蛋白的磷酸化，可以有效地抑制JCV感染。在未来，该药可能适用于JCV感染的进行性多灶性白质脑病病例。我们还以JCV Agno为诱饵，采用酵母双杂交实验研究了JCV病毒粒子在细胞内易位的机制。我们鉴定了两种细胞蛋白为agno结合蛋白。这些蛋白在JCV病毒粒子的核输出中起关键作用。这些结果被建立（EMBO Rep 6: 452, 2005和J Biol Chem 280,24948, 2005）。我们还建立了一些关于病毒感染与宿主反应相互作用的文章。关键字

In this project, we have attempted to inhibit JC virus (JCV) infection using small interfering RNAs (siRNAs) against JCV agnoprotein (Agno) as therapeutic strategy. In order to inhibit JCV infection in infected cells, we synthesized double-stranded siRNAs which are specific for JCV coding protein, Agno and introduced these into infected human glial-derived cells. We have observed that in post-infection treatment with siRNAs targeting JCV Agno suppresses virus production in JCV infected cells. Based on the results, we applied two patents and established the paper (J Virol 78: 7270, 2004). In addition, we have established JCV infection mimicking model using mice which were intracerebrally inoculated JCVinfected cells, and applied siRNA against JCV Agno to the model in vivo. After treatment with the siRNA, the number of intracerebral JCVinfected cells was significantly decreased. In addition, we have found that clinical drug was effectively suppressed JCV infection by modification of phosphorylation of a protein in vitro. In the future, the drug might be applicable for the JCV infected progressive multifocal leukoencephalopathy cases. We also examined the mechanism of intracellular translocation of JCV virion using the yeast two-hybrid assay using JCV Agno as a bait. We identified two cellular proteins as Agno-binding proteins. These proteins played a pivotal role in nuclear egress of JCV virion. These results were established (EMBO Rep 6: 452, 2005 and J Biol Chem 280, 24948, 2005). We also established some articles concerning about interaction between viral infection and host response.10. KEYWORDS