Establishment of induction method for Insulin producing cells by overexpression of large Maf transcription factors

我们检查了体内生物发光成像(BLI)是否可以监测胰腺外组织，特别是肝脏中胰岛素启动子活性的动态变化。为此，在小鼠胰岛素基因启动子的控制下，将荧光素酶报告基因表达的MIP-Luc小鼠的肝细胞，用表达胰岛素转录相关转录因子Pdx1、NeuD和MafA或MafB的重组腺病毒进行体内转导。在感染了所有这三种基因的小鼠身上，直到感染大约一周后，才检测到来自肝脏的生物发光信号。Pdx1、NeuroD和MafA重组腺病毒联合感染肝脏的免疫组织化学分析显示，一些胰岛素阳性细胞的存在与增强的BLI信号强度一致。这些结果表明，MIP-Luc小鼠肝区发出的BLI信号可用于监测胰岛素启动子的活性，Pdx1、Neurod和MafA具有诱导胰岛素转录的活性。

We examined whether in vivo bioluminescence imaging (BLI) could monitor the dynamics of insulin promoter activity in extrapancreatic tissues, especially in the liver. To do this, hepatocytes of MIP-Luc mice, in which luciferase reporter gene expresses under the control of mouse insulin gene promoter, were transduced in vivo with recombinant adenovirus expressing defined transcription factors related to insulin transcription, Pdx1, NeuroD, and MafA or MafB. The bioluminescent signal from the liver was detected in mice infected with all of three genes until about a week after the infection. Immunohistochemical analysis of the liver infected with combination of Pdx1, NeuroD, and MafA recombinant adenoviruses revealed the presence of some insulin positive cells consistent with the enhanced BLI signal intensity. These results suggest that the BLI signal emitted from the liver region of the MIP-Luc mice is useful for the monitoring the insulin promoter activity and Pdx1, NeuroD, and MafA have activity to induce insulin transcription.