Adenovirus-mediated transfer of a modified human proinsulin gene reverses hyperglycemia in diabetic mice.
Adenovirus-mediated transfer of a modified human proinsulin gene reverses hyperglycemia in diabetic mice.
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腺病毒介导的修饰人胰岛素原基因的转移可逆转糖尿病小鼠的高血糖。
DOI：
10.1152/ajpendo.1998.275.5.e748
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Pessin,JE
中科院分区：
文献类型：
--
作者：
Short,DK;Okada,S;Yamauchi,K;Pessin,JE
将人胰岛素原cDNA引入复制缺陷型腺病毒中，发现其在感染后可使肝细胞（H4-II-E）表达胰岛素原。构建第二种病毒，其中二元激素原转化酶识别序列突变为四元弗林蛋白酶切割位点。感染该病毒的细胞合成胰岛素原和成熟胰岛素。凝胶过滤色谱法、胰岛素结合的竞争和胰岛素受体激酶活性的活化证明，该成熟胰岛素与真实加工胰岛素在功能上相同。将这些病毒构建体注射到小鼠的颈外静脉中导致胰岛素基因在肝脏中表达。从突变的胰岛素原病毒的表达显着改善糖尿病小鼠的血糖状态。然而，病毒感染的影响是短暂的，在15 -7天达到最大值，并在14-21天恢复到稳态水平。这些数据表明，通过使用重组腺病毒进行体细胞胰岛素基因递送可用于短暂逆转小鼠的糖尿病状态。
The human proinsulin cDNA was introduced into a replication-defective adenovirus and was found to confer proinsulin expression to a hepatocyte (H4-II-E) cell line upon infection. A second virus was constructed in which the dibasic prohormone convertase recognition sequence was mutated to a tetrabasic furin cleavage site. Cells infected with this virus synthesized both proinsulin and mature insulin. Gel filtration chromatography, competition of insulin binding, and activation of the insulin receptor kinase activity demonstrated that this mature insulin was functionally identical to that of authentic processed insulin. Injection of these viral constructs into the external jugular vein of mice resulted in insulin gene expression in the liver. Expression from the mutated proinsulin virus dramatically improved the glycemic state of diabetic mice. However, the effects of the viral infection were transient, being maximal at ∼5–7 days and returning to steady-state levels by 14–21 days. These data demonstrate that somatic cell insulin gene delivery by the use of recombinant adenovirus can be used to transiently reverse the diabetic state in mice.