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Gene therapy of Hepatitis C using monkey by siRNA

使用猴子进行 siRNA 丙型肝炎基因治疗

基本信息

批准号：
16390204
负责人：
YOKOTA Takanori
金额：
$ 9.15万
依托单位：
Department of Neurology and Neurological Science, Tokyo medicak and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390204/
关键词：
RNA interference siRNA cationic vector Hepatitis C tamarin marmoset GBV-B interferon インターフェロン HCV

项目摘要

We synthesized the siRNA oligonucleotides targeting to the of GBV-B genome which is different from that of the corresponding sequence of HCV genome by only two nucleotides. This site was selected from 5' untranslated region, the most conservative region in the GBV-B/HCV genome, making the siRNA resistant to the mutation of the virus. For the delivery of the siRNA to the liver, we have developed a cationic liposome, LIC-101, which consists of new novel cationic lipid analogue.The five marmosets were used in this study. The 1.0, 2.5 and 5.0mg/kg of siRNA to GBV-B formulated by LIC-101 were slowly administered from the saphenous vein of the marmosets for three days. On the second day, GBV-B virus particles were directly injected to the liver. We could observe the dramatic and dose-dependent effect of the GBV-B siRNA/LIC101 to inhibit the replication GBV-B virus ; The 5.0mg/kg siRNA/LIC-101 completely suppressed the replication of GBV-B for more than 4 months after inoculation of the virus, even though the siRNA was injected for only three days. Unexpectedly, the control siRNA/LIC-101also can delay the virus replication. Since the control siRNA/LIC-101 induced the serum interferons α and γ, this delay of virus replication by the control siRNA/LIC-101 on the viral replication should be caused by an anti-viral effect of the induced interferons. In contrast, GBV-B siRNA/LIC-101 induced much less interferons than the control siRNA/LIC-101. Therefore, inhibitory effect of GBV-B siRNA/LIC-101 was considered to include those by both siRNA and interferons. Since synthetic siRNA formulated with lipid vectors is recognized through toll-like receptors located in the endosomal membrane and activates immune system in a sequence dependent manner.In conclusion, we succeeded to inhibit replication of GBV-B in liver of monkey with siRNA. We are trying the new method to suppress interferon response to escape the side effect.

我们合成了针对GBV-B基因组的siRNA寡核苷酸，其与HCV基因组的相应序列仅相差两个核苷酸。该位点选自GBV-B/HCV基因组中最保守的5'非翻译区，使siRNA能够抵抗病毒的突变。为了将siRNA递送到肝脏，我们开发了一种由新型阳离子脂质类似物组成的阳离子脂质体LIC-101。将LIC-101配制的1.0、2.5和5.0mg/kg抗GBV-B的SiRNA从绒猴隐静脉缓慢给药三天。第二天，将GBV-B病毒颗粒直接注射到肝脏中。结果表明，GBV-B siRNA/LIC-101对GBV-B病毒的复制有明显的抑制作用，且呈剂量依赖性; 5.0mg/kg的siRNA/LIC-101对GBV-B病毒的复制有明显的抑制作用，即使只注射3天，也能持续4个月以上。出乎意料的是，对照siRNA/LIC-101也可以延迟病毒的复制。由于对照siRNA/LIC-101诱导血清干扰素α和γ，因此对照siRNA/LIC-101对病毒复制的延迟应该是由诱导的干扰素的抗病毒作用引起的。相比之下，GBV-B siRNA/LIC-101比对照siRNA/LIC-101诱导少得多的干扰素。因此，认为GBV-B siRNA/LIC-101的抑制作用包括siRNA和干扰素的抑制作用。由于合成的siRNA与脂质载体一起配制，通过位于内体膜上的toll样受体识别，并以序列依赖的方式激活免疫系统。总之，我们成功地用siRNA抑制了GBV-B在猴肝脏中的复制。我们正在尝试新的方法来抑制干扰素的反应，以逃避副作用。