Using the Adeno-associated virus (AAV) as a gene delivery vehicle, we have constructed a recombinant vector containing the full length rat preproinsulin gene (vLP-1). Utilizing the well described non-obese diabetic (NOD) mouse model, an experimental group (n=10) of animals were intramuscularly (IM) injected with107rAAV virions containing the insulin gene and compared to a mock-injected control group (n=10). Blood glucose (glc) was then measured weekly for 16 weeks. Data showed that the experimental group contained 70% euglycemic animals (defined as glc <200mg/dL)versus10% of the control animals (P<.05) at 14 weeks. Mean weight in the treated group was greater than the untreated group. Insulin mRNA was detected at the injection site of all of the treated animals, but not controls. Complete destruction of islets was confirmed by histology ruling out the possibility of spontaneous reversal of insulinitis. We conclude that IM delivery of the insulin gene in the NOD mouse was able to prevent clinical DM up to 14 weeks in a majority of treated animals. Our experimental data suggests that gene therapy may be an alternative treatment for IDDM in the future.
我们利用腺相关病毒(AAV)作为基因递送载体,构建了一种含有全长大鼠前胰岛素基因(vLP-1)的重组载体。利用已被充分描述的非肥胖糖尿病(NOD)小鼠模型,向实验组(n=10)动物肌肉注射含有胰岛素基因的 107rAAV 病毒,并与模拟注射对照组(n=10)进行比较。然后每周测量血糖(glc),持续 16 周。数据显示,14 周时,实验组有 70% 的动物出现优生(定义为 glc <200mg/dL),而对照组只有 10% 的动物出现优生(P<.05)。治疗组的平均体重高于未治疗组。在所有治疗组动物的注射部位都检测到了胰岛素 mRNA,而对照组没有检测到。组织学检查证实了胰岛的完全破坏,排除了胰岛素炎自发逆转的可能性。我们得出的结论是,在 NOD 小鼠体内注射胰岛素基因能够预防大多数治疗动物的临床 DM,最长可达 14 周。我们的实验数据表明,基因疗法可能是未来治疗 IDDM 的一种替代疗法。