Lee So Hyun, Rhee Marie, Kim Ji Won, Yoon Kun Ho
Division of Endocrinology and Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea.
Diabetes Metab J. 2017 Oct;41(5):405-416. doi: 10.4093/dmj.2017.41.5.405.
To develop surrogate insulin-producing cells for diabetes therapy, adult stem cells have been identified in various tissues and studied for their conversion into β-cells. Pancreatic progenitor cells are derived from the endodermal epithelium and formed in a manner similar to gut progenitor cells. Here, we generated insulin-producing cells from the intestinal epithelial cells that induced many of the specific pancreatic transcription factors using adenoviral vectors carrying three genes: PMB (pancreatic and duodenal homeobox 1 [Pdx1], V-maf musculoaponeurotic fibrosarcoma oncogene homolog A [MafA], and BETA2/NeuroD).
By direct injection into the intestine through the cranial mesenteric artery, adenoviruses (Ad) were successfully delivered to the entire intestine. After virus injection, we could confirm that the small intestine of the mouse was appropriately infected with the Ad-Pdx1 and triple Ad-PMB.
Four weeks after the injection, insulin mRNA was expressed in the small intestine, and the insulin gene expression was induced in Ad-Pdx1 and Ad-PMB compared to control Ad-green fluorescent protein. In addition, the conversion of intestinal cells into insulin-expressing cells was detected in parts of the crypts and villi located in the small intestine.
These data indicated that PMB facilitate the differentiation of mouse intestinal cells into insulin-expressing cells. In conclusion, the small intestine is an accessible and abundant source of surrogate insulin-producing cells.
为了开发用于糖尿病治疗的替代胰岛素产生细胞,已在各种组织中鉴定出成体干细胞,并对其向β细胞的转化进行了研究。胰腺祖细胞来源于内胚层上皮,其形成方式与肠道祖细胞相似。在此,我们利用携带三个基因(PMB,即胰腺和十二指肠同源盒1 [Pdx1]、V-maf 肌腱膜纤维肉瘤癌基因同源物A [MafA]和BETA2/NeuroD)的腺病毒载体,从诱导出许多特定胰腺转录因子的肠上皮细胞中生成了胰岛素产生细胞。
通过肠系膜上动脉直接将腺病毒注射到肠道中,成功地将腺病毒递送至整个肠道。病毒注射后,我们能够确认小鼠的小肠被Ad-Pdx1和三联Ad-PMB适当感染。
注射四周后,胰岛素mRNA在小肠中表达,与对照Ad-绿色荧光蛋白相比,Ad-Pdx1和Ad-PMB诱导了胰岛素基因表达。此外,在小肠隐窝和绒毛的部分区域检测到肠细胞向表达胰岛素细胞的转化。
这些数据表明PMB促进小鼠肠细胞向表达胰岛素细胞的分化。总之,小肠是替代胰岛素产生细胞的一个易于获取且丰富的来源。