Center of Animal Biotechnology and Gene Therapy, Universitat Autònoma Barcelona, Bellaterra, Spain.
Diabetes. 2013 May;62(5):1718-29. doi: 10.2337/db12-1113. Epub 2013 Feb 1.
Diabetes is associated with severe secondary complications, largely caused by poor glycemic control. Treatment with exogenous insulin fails to prevent these complications completely, leading to significant morbidity and mortality. We previously demonstrated that it is possible to generate a "glucose sensor" in skeletal muscle through coexpression of glucokinase and insulin, increasing glucose uptake and correcting hyperglycemia in diabetic mice. Here, we demonstrate long-term efficacy of this approach in a large animal model of diabetes. A one-time intramuscular administration of adeno-associated viral vectors of serotype 1 encoding for glucokinase and insulin in diabetic dogs resulted in normalization of fasting glycemia, accelerated disposal of glucose after oral challenge, and no episodes of hypoglycemia during exercise for >4 years after gene transfer. This was associated with recovery of body weight, reduced glycosylated plasma proteins levels, and long-term survival without secondary complications. Conversely, exogenous insulin or gene transfer for insulin or glucokinase alone failed to achieve complete correction of diabetes, indicating that the synergistic action of insulin and glucokinase is needed for full therapeutic effect. This study provides the first proof-of-concept in a large animal model for a gene transfer approach to treat diabetes.
糖尿病与严重的继发性并发症有关，这些并发症主要是由血糖控制不佳引起的。外源性胰岛素治疗并不能完全预防这些并发症，导致发病率和死亡率显著增加。我们之前的研究表明，通过共表达葡萄糖激酶和胰岛素，在骨骼肌中产生“葡萄糖传感器”是可能的，这可以增加葡萄糖摄取并纠正糖尿病小鼠的高血糖。在这里，我们在糖尿病的大型动物模型中证明了这种方法的长期疗效。在糖尿病犬中一次性肌内给予血清型 1 的腺相关病毒载体，可使空腹血糖正常化，口服葡萄糖负荷后葡萄糖清除加速，且在基因转移后 >4 年的运动过程中无低血糖发作。这与体重恢复、糖化血浆蛋白水平降低以及无继发性并发症的长期存活有关。相反，外源性胰岛素或单独转胰岛素或葡萄糖激酶的基因治疗未能完全纠正糖尿病，表明胰岛素和葡萄糖激酶的协同作用对于达到完全的治疗效果是必需的。这项研究在大型动物模型中首次提供了基因治疗方法治疗糖尿病的概念验证。
