人源化小鼠模型中，原代人肝细胞的部分胰腺转分化。

Partial pancreatic transdifferentiation of primary human hepatocytes in the livers of a humanised mouse model.

机构信息

School of Life Sciences, University of Technology Sydney, Sydney, Australia.

The Centre for Health Technologies, University of Technology Sydney, Sydney, Australia.

出版信息

J Gene Med. 2018 May;20(5):e3017. doi: 10.1002/jgm.3017. Epub 2018 Apr 16.

DOI:10.1002/jgm.3017

PMID:29578255

Abstract

BACKGROUND

Gene therapy is one treatment that may ultimately cure type 1 diabetes. We have previously shown that the introduction of furin-cleavable human insulin (INS-FUR) to the livers in several animal models of diabetes resulted in the reversal of diabetes and partial pancreatic transdifferentiation of liver cells. The present study investigated whether streptozotocin-diabetes could be reversed in FRG mice in which chimeric mouse-human livers can readily be established and, in addition, whether pancreatic transdifferentiation occurred in the engrafted human hepatocytes.

METHODS

Engraftment of human hepatocytes was confirmed by measuring human albumin levels. Following delivery of the empty vector or the INS-FUR vector to diabetic FRG mice, mice were monitored for weight and blood glucose levels. Intraperitoneal glucose tolerance tests (IPGTTs) were performed. Expression levels of pancreatic hormones and transcription factors were determined by a reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry.

RESULTS

Diabetes was reversed for a period of 60 days (experimental endpoint) after transduction with INS-FUR. IPGTTs of the insulin-transduced animals were not significantly different from nondiabetic animals. Immunofluorescence microscopy revealed the expression of human albumin and insulin in transduced liver samples. Quantitative RT-PCR showed expression of human and mouse endocrine hormones and β-cell transcription factors, indicating partial pancreatic transdifferentiation of mouse and human hepatocytes. Nonfasting human C-peptide levels were significantly higher than mouse levels, suggesting that transdifferentiated human hepatocytes made a significant contribution to the reversal of diabetes.

CONCLUSIONS

These data show that human hepatocytes can be induced to undergo partial pancreatic transdifferentiation in vivo, indicating that the technology holds promise for the treatment of type 1 diabetes.

摘要

背景

基因治疗是一种可能最终治愈 1 型糖尿病的治疗方法。我们之前已经表明，在几种糖尿病动物模型中，将可被弗林蛋白酶切割的人胰岛素（INS-FUR）引入肝脏，可以逆转糖尿病并使肝细胞发生部分胰腺转分化。本研究调查了嵌合鼠人肝脏容易建立的 FRG 小鼠中是否可以逆转链脲佐菌素诱导的糖尿病，并且，在移植的人肝细胞中是否发生胰腺转分化。

方法

通过测量人白蛋白水平来确认人肝细胞的移植。在将空载体或 INS-FUR 载体递送至糖尿病 FRG 小鼠后，监测小鼠的体重和血糖水平。进行腹腔内葡萄糖耐量试验（IPGTTs）。通过逆转录聚合酶链反应（RT-PCR）和免疫组织化学法来确定胰腺激素和转录因子的表达水平。

结果

在用 INS-FUR 转导后，糖尿病在 60 天（实验终点）内得到逆转。经胰岛素转导的动物的 IPGTT 与非糖尿病动物没有明显差异。免疫荧光显微镜显示转导的肝样本中表达了人白蛋白和胰岛素。定量 RT-PCR 显示了人源和鼠源内分泌激素和β细胞转录因子的表达，表明鼠源和人源肝细胞发生了部分胰腺转分化。非禁食状态下人 C 肽水平明显高于鼠源水平，表明转分化的人肝细胞对糖尿病的逆转有重要贡献。