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.

我们合成了靶向GBV-B基因组的siRNA寡核苷酸，该siRNA寡核苷酸与仅通过两个核苷酸的相应HCV基因组序列不同。该位点是从5'非翻译区域中选择的，该区域是GBV-B/HCV基因组中最保守的区域，使siRNA具有耐药性的病毒突变。为了将siRNA递送到肝脏，我们开发了一个阳离子脂质体LIC-101，该脂质体由新的新型阳离子脂质类似物组成。这项研究中使用了五根摩尔果。 LIC-101制定的1.0、2.5和5.0mg/kg的siRNA至GBV-B从果marmoset的大静脉缓慢施用了三天。第二天，将GBV-B病毒颗粒直接注入肝脏。我们可以观察GBV-B SIRNA/LIC101的戏剧性和剂量依赖性作用，以抑制复制GBV-B病毒； 5.0mg/kg siRNA/LIC-101在接种病毒后4个月以上，完全抑制了GBV-B的复制，即使仅注射siRNA三天。出乎意料的是，控制siRNA/LIC-101会延迟病毒复制。由于对照siRNA/LIC-101诱导了血清干扰素α和γ，因此对控制siRNA/LIC-101对病毒复制的这种病毒复制的延迟应是由诱导中断的抗病毒作用引起的。相比之下，GBV-B SIRNA/LIC-101的干扰少于对照siRNA/LIC-101。因此，GBV-B siRNA/LIC-101的抑制作用被认为包括siRNA和干扰的抑制作用。由于用脂质载体制成的合成siRNA通过位于内体膜中的Toll样受体识别，并以序列依赖性的方式激活免疫系统。总之，我们成功地抑制了用Sirne的猴子肝脏中GBV-B的复制。我们正在尝试采用新方法来抑制干扰响应以逃避副作用。