Nukuzuma Souichi, Nakamichi Kazuo, Kameoka Masanori, Sugiura Shigeki, Nukuzuma Chiyoko, Tasaki Takafumi, Takegami Tsutomu
Department of Infectious Diseases, Kobe Institute of Health, 4-6-5, Minatojima-Nakamachi, Chuo-ku, Kobe 650-0046.
Department of Virology 1, National Institute of Infectious Diseases, Toyama, Shinjuku, Tokyo 162-8640.
Microbiol Immunol. 2016 Apr;60(4):253-60. doi: 10.1111/1348-0421.12372.
JC polyomavirus (JCPyV) causes progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease of the central nervous system, in immunocompromised patients. Because no drugs have been approved for treating PML, many antiviral agents are currently being investigated for this purpose. The inhibitory effects of the topoisomerase I inhibitors topotecan and β-lapachone were assessed by investigating viral replication, propagation and viral protein 1 (VP1) production in cultured cells. JCPyV replication was assayed using the human neuroblastoma cell line IMR-32 transfected with the JCPyV plasmid and RT- PCR combined with Dpn I treatment. Dpn I digests the input plasmid DNA containing methylated adenosine, but not newly replicated JCPyV DNA, in IMR-32 cells. It was found that JCPyV replicates less in IMR-32 cells treated with topotecan or β-lapachone than in untreated cells. Moreover, drug treatment of JCI cells, which are IMR-32 cells persistently infected with JCPyV, led to a reduction in the amount of JCPyV DNA and population of VP1-positive cells. These results demonstrate that topotecan and β-lapachone affects JCPyV propagation in human neuroblastoma cell lines, suggesting that topotecan and β-lapachone could potentially be used to treat PML.
JC多瘤病毒（JCPyV）可在免疫功能低下的患者中引发进行性多灶性白质脑病（PML），这是一种致命的中枢神经系统脱髓鞘疾病。由于尚无获批用于治疗PML的药物，目前许多抗病毒药物正为此接受研究。通过研究培养细胞中的病毒复制、传播及病毒蛋白1（VP1）生成情况，评估了拓扑异构酶I抑制剂拓扑替康和β-拉帕醌的抑制作用。使用转染了JCPyV质粒的人神经母细胞瘤细胞系IMR-32，并结合Dpn I处理的逆转录聚合酶链反应（RT-PCR）来检测JCPyV复制情况。Dpn I可消化IMR-32细胞中含有甲基化腺苷的输入质粒DNA，但不会消化新复制的JCPyV DNA。结果发现，与未处理的细胞相比，用拓扑替康或β-拉帕醌处理的IMR-32细胞中JCPyV的复制较少。此外，对持续感染JCPyV的IMR-32细胞系JCI细胞进行药物处理，可导致JCPyV DNA量及VP1阳性细胞数量减少。这些结果表明，拓扑替康和β-拉帕醌会影响JCPyV在人神经母细胞瘤细胞系中的传播，提示拓扑替康和β-拉帕醌可能有潜力用于治疗PML。
