NeuroD/betacellulin gene therapy induces islet neogenesis in the liver and reverses diabetes in mice

据报道，第一代腺病毒（FGAd）介导的同源异型盒基因Pdx-1的转移可诱导链脲佐菌素（STZ）治疗的糖尿病小鼠的肝脏胰岛素产生和短暂的低血糖。与其效果是短暂的FG-广告，辅助依赖性（HD）广告产生延长的转基因表达。我们使用HDAd将Pdx-1递送至STZ小鼠，并观察到由胰蛋白酶表达引起的高血糖症的部分改善，但严重的、致命的肝炎，这是由Pdx-1的外分泌分化作用引起的。HDAd介导的NeuroD（Pdx-1下游的一种发育因子）转移部分逆转了STZ小鼠的高血糖，而不诱导肝炎。在NeuroD中添加胰岛生长因子β细胞素完全纠正了糖尿病。治疗的小鼠在实验期间（>120天）是健康的和血糖正常的。我们在肝脏中检测到胰岛素和其他胰岛特异性转录物，包括胰岛素原加工酶、β细胞特异性葡萄糖激酶和磺酰脲受体。免疫组化显示存在胰岛素，胰高血糖素，胰多肽（PP）和生长抑素产生细胞组织成胰岛簇，免疫电镜显示典型的胰岛素特异性颗粒。我们的数据表明，NeuroD/betacellulin是一个有前途的基因治疗方案，用于治疗胰岛素依赖型糖尿病。这些结果发表在Nat.Med9，596，2003和许多评论文章中，如出版记录所示。

First-generation adenovirus(FGAd)-mediated transfer of the homeobox gene Pdx-1 was reported to induce hepatic insulin production and transient hypoglycemia in streptozotocin(STZ)-treated diabetic mice. Unlike FG-Ads whose effects are short-lived, helper-dependent(HD) Ads produce prolonged transgene expression. We used HDAd to deliver Pdx-1 to STZ mice and observed partial amelioration of hyperglycemia, but severe, fatal, hepatitis, caused by expression of trypsin, resulting from the exocrine-differentiating action of Pdx-1. HDAd-mediated transfer of NeuroD, a developmental factor downstream of Pdx-1,partially reversed the hyperglycemia of STZ mice without inducing hepatitis. The addition of an islet growth factor betacellulin to NeuroD completely corrected the diabetes. Treated mice were healthy and normoglycemic for the duration of the experiment (>120 days). We detected in the liver insulin and other islet-specific transcripts, including proinsulin-processing enzymes, beta cell-specific glucokinase, and sulfonylurea receptor. Immunohistochemistry revealed the presence of insulin, glucagon, pancreatic polypeptide(PP) and somatostatin-producing cells organized into islet clusters, and immuno-electron microscopy showed typical insulin-specific granules. Our data suggest that NeuroD/betacellulin is a promising gene therapy regimen for the treatment of insulin-dependent diabetes. These results were published in Nat. Med 9,596,2003 and many review articles as shown in the publication records.