ADENOVIRAL MEDIATED ISLET GENE TRANSFER

糖尿病是一种常见疾病，估计有5%的人会患上糖尿病 占美国总人口的比例。糖尿病是糖尿病的一个主要原因 血管疾病过早死亡是致盲的主要原因 在工作年龄的成年人中，非创伤性的下肢截肢，以及 肾功能衰竭需要透析和移植。约10-15 美国医疗保健总预算的10%花在糖尿病上。 因此，糖尿病对个人和 社会。糖尿病(I型和II型)发病机制的研究 表明贝塔细胞功能受损占大多数 这些缺陷。虽然这两种疾病的发病机制是 不同的是，这两种糖尿病的潜在代谢原因是 胰岛β细胞不能提供一定数量的胰岛素 足以满足身体的需要，导致高血糖。 控制不当的高血糖直接导致了 特异型和非特异型并发症的研究进展 糖尿病状态。所有糖尿病患者都存在β细胞功能障碍 病人。同种异体胰岛移植物也可能出现β细胞功能障碍。在……里面 葡萄糖调节的β细胞功能、葡萄糖摄取和代谢 在发送消息时必须采取的步骤，从而导致 细胞内钙离子，导致胰岛素的生物合成和分泌。这些 过程受两种蛋白质的直接调节：葡萄糖转运体 和葡萄糖激活剂。有人提出，GLUT2和葡萄糖激酶可能 作为葡萄糖传感仪器的主要元件而串联运行。 通过在胰岛和胰岛β细胞中过表达这些基因 LINES，已经证实胰岛素释放显著增加 在葡萄糖诱导的反应中。然而，这一现象并没有 用人的胰岛进行了研究。在这项提案中，我们将 通过以下途径将这些候选基因(S)导入人胰岛组织 血管递送腺病毒载体并检测其是否具有特异性 基因转移可改善胰岛β细胞功能 活着。具体地说，我们将进一步定义适当的条件来 实现高效(90%以上)的基因转移 经血管输送病毒的人胰岛组织 (动脉灌注)胰岛前的全胰腺灌注 与世隔绝。然后我们将评估病毒感染对新陈代谢的影响 和基因转移(第一、第二、第三代腺病毒载体编码 LacZ报告基因或其他特异性治疗基因(S) 胰岛组织的功能完整性。最后，我们将研究 腺病毒介导的胰岛基因转移的实验研究 I型胰岛β细胞功能障碍的矫治 和II糖尿病通过将GLUT2和葡萄糖激酶基因转移到 人的胰岛组织。

Diabetes mellitus is a common disorder, estimated to occur in 5 percent of the population of the United States. Diabetes is a major cause of premature death from vascular disease and the leading cause of blindness in working-age adults, of nontraumatic lower extremity amputations, and of renal failure requiring dialysis and transplantation. About 10-15 percent of the total health care budget in the US is spent on diabetes. Thus, diabetes represents a serious burden to the individual and to society. Studies on the pathogenesis of diabetes (both type I and II) indicate that impaired Beta-cell function accounts for the majority of the defects. Although the pathogenesis of the two diseases is different, the underlying metabolic cause of both forms of diabetes is a failure of the pancreatic Beta cells to provide insulin in an amount sufficient to meet the body's needs, leading to hyperglycemia. Improperly controlled hyperglycemia is directly responsible for the development of the specific and non-specific complications of the diabetic state. Beta-cell dysfunction is present in all diabetic patients. Islet allografts may also exhibit Beta-cell dysfunction. In glucose-regulated Beta-cell function, glucose uptake and metabolism are obligatory steps in sending the message which results in an increase in intracellular Ca2+, leading to insulin biosynthesis and secretion. These processes are directly regulated to two proteins: glucose transporters and glucokinase. It was proposed that GLUT2 and glucokinase may function in tandem as primary elements of the glucose sensing apparatus. By overexpression of these genes in pancreatic islets and Beta-cell lines, a significant increase of insulin release has been demonstrated in glucose-induced response. However, this phenomenon has not been studied using human pancreatic islets. In this proposal, we will transfer these candidate gene(s) into human pancreatic islet tissue by vascular delivery of adenoviral vectors and examine whether specific gene transfer will improve islet Beta-cell function in vitro and in vivo. Specifically, we will further define appropriate conditions to achieve highly efficient (greater than 90 percent) gene transfer to human pancreatic islet tissue by delivering viruses through vascular (arterial infusion) perfusion of the whole pancreas prior to islet isolation. We will then assess the metabolic effects of viral infection and gene transfer (1st, 2nd, 3rd generation adenoviral vectors encoding LacZ reporter gene or other specific therapeutic gene(s) on the functional integrity of islet tissue. Finally we will examine the effectiveness of adenoviral mediated gene transfer to pancreatic islets on the correction of Beta-cell dysfunction for the treatment of Type I and II diabetes by transferring the genes for GLUT2 and glucokinase to human islet tissue.