权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Targetting of the brain by cultured microglia cell for neurofrgenerative disorders.

通过培养的小胶质细胞靶向大脑治疗神经再生障碍。

基本信息

批准号：
11557060
负责人：
TANAKA Akemi
金额：
$ 5.63万
依托单位：
Osaka City University(Graduate School of Medicine)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

Gene delivery into the brain via blood vessels is quite difficult because of the blood-brain-barrier (BBB). In the literature, a number of experiments of gene therapy for the brain have been done, but none of them has been successful. As it is speculated that microglia cells would go through BBB, we established a strain of cultured microglia cell from newborn mice brain for the vehicle of gene into the brain. The cells were labeled with a reporter gene of GFP and injected into the left ventricle of the heart of Sly mice. Sly mouse, the deficiency of β-glucuronidase, is a mouse model for human disease of mucopolysaccharidosis type VII, which is a systemic disorder including the brain caused by the accumulation of glycosaminoglycans. It is suggested that delivery of the deficient enzyme or the gene into each organ of this model mouse would improve the disease.Our results showed that the exogenous microglia cells were seen only in the brain of the mice with the advanced stage of the disease, or in the ischemic brain. Thus, the cultured microglia cells could enter the brain though BBB only when BBB was injured. They could not found in the normal brain. However, when adenovirus vector with β-galactosidase gene was injected into the temporal surface vein of newborn mice with β-galactosidase deficiency, virus vectors could enter the brain and show β-galactosidase activity. Moreover, the accumulation of GM2 ganglioside was suppressed.

由于血脑屏障（BBB）的存在，基因通过血管进入大脑是相当困难的。在文献中，已经进行了许多大脑基因治疗的实验，但没有一个是成功的。由于推测小胶质细胞会通过血脑屏障，我们从新生小鼠的大脑中培养了一株小胶质细胞作为基因进入大脑的载体。这些细胞被GFP报告基因标记，并注射到Sly小鼠的左心室。β-葡萄糖醛酸酶缺乏的Sly小鼠是人类粘多糖病VII型的小鼠模型，粘多糖病是一种由糖胺聚糖积累引起的包括大脑在内的全身性疾病。这表明，将缺陷酶或基因输送到模型小鼠的每个器官将改善疾病。我们的结果表明，外源性小胶质细胞仅在疾病晚期小鼠的大脑中或在缺血性大脑中可见。因此，培养的小胶质细胞只有在血脑屏障损伤时才能通过血脑屏障进入大脑。它们在正常大脑中是找不到的。然而，将含有β-半乳糖苷酶基因的腺病毒载体注入β-半乳糖苷酶缺乏的新生小鼠颞面静脉后，病毒载体可以进入大脑并显示出β-半乳糖苷酶活性。此外，GM2神经节苷脂的积累受到抑制。