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Development of basic techniques for the liver-directed gene therapy

肝脏定向基因治疗基础技术的开发

基本信息

批准号：
07557071
负责人：
ISHIBASHI Shun
金额：
$ 9.47万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

Liver-directed gene therapy may be an ultimate therapy for many genetic diseases such as familial hypercholesterolemia (FH). Direct introduction of low density lipoprotein receptor (LDLR) expression into null type of FH may potentially cause antibody formation against the introduced protein, and mitigate the therapeutic effects. To circumbent this problem, we have tested the feasibility of using non- LDLR proteins such as lipoprotein lipase (LPL) and apoliporptein E (apoE). Both proteins may function as ligands for the non-LDLR pathway for hepatic lipoprotein catabolism. We have generated two types of mice : i) LDLR knockout mice overexpressiong LPL under the control of CAG promoter (LPLTg ; LDLRKO), ii) LDLR knockout mice overexpressiong rat apoE under the control of methanotionein promoter (ETg ; LDLRKO). In both animals, cholesterol-lowering effects were observed. More importantly, diet-induced atherosclerosis was significantly suppressed in these animals. These results suggest that LPL or apoE are promising candidate genes as a surrogate for LDLR.The other approach is ex vivo gene therapy. However, its limitation is that the hepatocytes transplanted to the recipient liver do not regenerate to the level which is enough to rescue the metabolic defects. As an experimental tool to investigate the recipient hepatocyte-specific cell ablation, we have generated mice lacking asialoglycoprotein receptor (ASGPR). In the current study, HL1, a major component of ASGPR, has been disrupted. HSL-/- mice lack both HL1 and HL2 in the liver. Plasma clearance of ASGP was almost completely blocked.

肝靶向基因治疗可能是家族性高胆固醇血症（FH）等许多遗传性疾病的最终治疗方法。将低密度脂蛋白受体（LDLR）直接导入空型FH中可能会导致针对所导入蛋白的抗体形成，并降低治疗效果。为了解决这个问题，我们已经测试了使用非LDLR蛋白质如脂蛋白脂酶（LPL）和载脂蛋白E（apoE）的可行性。这两种蛋白质都可以作为肝脂蛋白催化剂的非LDLR途径的配体。我们已经产生了两种类型的小鼠：i）在CAG启动子控制下过表达LPL的LDLR敲除小鼠（LPLTg ; LDLRKO），ii）在蛋氨酸启动子控制下过表达大鼠apoE的LDLR敲除小鼠（ETg ; LDLRKO）。在两只动物中均观察到了降胆固醇作用。更重要的是，饮食诱导的动脉粥样硬化在这些动物中被显著抑制。这些结果表明，LPL或apoE是有希望的候选基因作为替代LDLR。另一种方法是离体基因治疗。然而，其局限性在于移植到受体肝脏的肝细胞不能再生到足以挽救代谢缺陷的水平。作为研究受体肝细胞特异性细胞消融的实验工具，我们产生了缺乏去唾液酸糖蛋白受体（ASGPR）的小鼠。在目前的研究中，HL 1，ASGPR的主要组成部分，已被破坏。HSL-/-小鼠在肝脏中缺乏HL 1和HL 2。ASGP的血浆清除几乎完全被阻断。