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

在本项目中，我们尝试使用针对JC病毒（JCV）未知蛋白（阿尼奥）的小干扰RNA（siRNA）作为治疗策略来抑制JC病毒（JCV）感染。为了抑制感染细胞中的JCV感染，我们合成了对JCV编码蛋白阿尼奥特异性的双链siRNA，并将其引入感染的人神经胶质衍生细胞中。我们已经观察到，在感染后用靶向JCV阿尼奥的siRNA治疗抑制了JCV感染细胞中的病毒产生。基于此结果，我们申请了两项专利，并建立了论文（J Virol 78：7270，2004）。此外，我们还建立了脑内接种JCV感染细胞的小鼠JCV感染模拟模型，并将针对JCV阿尼奥的siRNA应用于体内模型。用siRNA处理后，脑内JCV感染细胞的数量显著减少。此外，我们还发现临床药物在体外通过修饰蛋白质的磷酸化而有效地抑制JCV感染。将来，该药可能适用于JCV感染的进行性多灶性白质脑病病例。我们还使用酵母双杂交试验，以JCV阿尼奥为诱饵，研究了JCV病毒粒子的细胞内易位机制。我们确定了两个细胞蛋白作为Agno结合蛋白。这些蛋白质在JCV病毒粒子的核出口中起关键作用。这些结果已经确立（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和宿主反应之间相互作用的文章。关键词

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