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treatment using adenovirus vector

使用腺病毒载体进行治疗

基本信息

批准号：
09357009
负责人：
OKA Yoshitomo
金额：
$ 17.09万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09357009/
关键词：
diabetes mellitus adenovirus vector insulin glucose transport PI3-kinase sulfonylurea receptor Wolfram syndrome persistent hyperinsulinemic hypoglycemia of infancy インスリン分泌膵β細胞遺伝子導入 hepatocyte nuclear factor 遺伝子治療 GKラットグリセロリン

项目摘要

We have studied insulin signaling toward stimulation of glucose transport in 3T3-L1 adipocytes using recombinant adenovirus. Expression of dominant negative p85 submit of PI3-kinase inhibited insulin-stimulated glucose transeport activity and GLUT4 translocation, indicating an important role of PI3-kinase activation in insulin-stimulated glucose transport. However, a discrepancy was observed between PI3-kinase activity and glucose transport activity, suggesting a possibility that a different pathway(s) is involved in insulin-stimulated intrinsic activity of glucose transport. In contrast to the involvement of PI3-kinase, ras activation is not involeved in insulin-stimulated glucose transeport, since overexpression of dominant negative Ras did not affect insulin-stimulated glucose transport.We have also studied insulin secretion mechanism using recombinant adenovirus. Mitochondrial FAD-linked glycerol-3-phosphate dehydrogenase (mGPDH) was decreased in pancreatic islets of GK rats which show defects in insulin serection. Since defects in insulin secretion had been ascribed to decreased mGPDH activity, we corrected the decreased mGPDH activity by overexpressing mGPDH using adenovirus vector. However, defects in insulin secretion was not corrected, indicating decreased mGPDH activity is not responsible for insulin secretion defects in GK rats. Employing positional cloning method, we have cloned a novel gene responsible for Wolfram syndrome characterized by non-immune insulin dependent diabetes mellitus and optic atrophy. This gene, designated WFS1, must have an important role in survival of pancreatic beta cells and neuronal cells. We also found mutations in sulfonylurea receptor gene in three Japanese patients with persistent hyperinsulinemic hypoglycemia of infancy and elucidated the molecular mechanism for defests in insulin secretion, especially a possible binding site of ADP and ATP for sulfonylurea receptor.

我们已经研究了胰岛素信号刺激葡萄糖转运在3 T3-L1脂肪细胞使用重组腺病毒。PI 3-激酶显性负性p85亚单位的表达抑制胰岛素刺激的葡萄糖转运活性和GLUT 4转运，表明PI 3-激酶激活在胰岛素刺激的葡萄糖转运中起重要作用。然而，在PI 3-激酶活性和葡萄糖转运活性之间观察到差异，表明不同途径参与胰岛素刺激的葡萄糖转运的内在活性的可能性。与PI 3-激酶的激活不同，显性负性Ras的过表达并不影响胰岛素刺激的葡萄糖转运，因此ras的激活并不参与胰岛素刺激的葡萄糖转运。GK大鼠胰岛线粒体FAD-连接的甘油-3-磷酸脱氢酶（mGPDH）减少，表现出胰岛素分泌缺陷。由于胰岛素分泌缺陷归因于mGPDH活性降低，我们通过使用腺病毒载体过表达mGPDH来校正mGPDH活性降低。然而，胰岛素分泌缺陷没有得到纠正，表明mGPDH活性降低不是GK大鼠胰岛素分泌缺陷的原因。我们采用定位克隆的方法，克隆了一个新的基因，该基因与以非免疫性胰岛素依赖型糖尿病和视神经萎缩为特征的Wolfram综合征有关。这个基因，命名为WFS 1，必须在胰腺β细胞和神经元细胞的存活中发挥重要作用。我们还在3例日本婴儿期持续性高胰岛素血症性低血糖患者中发现磺脲类药物受体基因突变，并阐明了胰岛素分泌障碍的分子机制，特别是磺脲类药物受体可能的ADP和ATP结合位点。