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

Binhai Ren; Que T La; Bronwyn A O'Brien; Najah T Nassif; Yi Tan; Dario Gerace; Rosetta Martiniello-Wilks; Fraser Torpy; Allison P Dane; Ian E Alexander; Ann M Simpson

doi:10.1002/jgm.3017

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

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

Authors

Binhai Ren^{1

2}, Que T La^{1

2}, Bronwyn A O'Brien^{1

2}, Najah T Nassif^{1

2}, Yi Tan¹, Dario Gerace^{1

2}, Rosetta Martiniello-Wilks^{1

2}, Fraser Torpy¹, Allison P Dane³, Ian E Alexander^{3

4}, Ann M Simpson^{1

2}

Affiliations

¹ School of Life Sciences, University of Technology Sydney, Sydney, Australia.
² The Centre for Health Technologies, University of Technology Sydney, Sydney, Australia.
³ The Children's Hospital at Westmead and Children's Medical Research Institute, Sydney, Australia.
⁴ Discipline of Child and Adolescent Health, University of Sydney, Sydney, Australia.

PMID: 29578255
DOI: 10.1002/jgm.3017

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.

Keywords: diabetes; gene expression; gene therapy; immunohistochemistry; liver; viral vector.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Transdifferentiation / genetics*
Cell Transplantation / methods
Cells, Cultured
Diabetes Mellitus, Experimental / genetics
Diabetes Mellitus, Experimental / therapy*
Diabetes Mellitus, Type 1 / genetics
Diabetes Mellitus, Type 1 / therapy*
Disease Models, Animal
Hepatocytes / cytology
Hepatocytes / metabolism*
Humans
Insulin / genetics
Insulin / metabolism*
Insulin-Secreting Cells / cytology
Insulin-Secreting Cells / metabolism*
Lentivirus / genetics
Liver / cytology
Liver / metabolism*
Mice
Transplantation, Heterologous

Substances

Insulin