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Diabetologia
Published in: Diabetologia 9/2007

Open Access 01-09-2007 | Article

Long-term correction of diabetes in rats after lentiviral hepatic insulin gene therapy

Authors: B. Ren, B. A. O’Brien, M. A. Swan, M. E. Koina, N. Nassif, M. Q. Wei, A. M. Simpson

Published in: Diabetologia | Issue 9/2007

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Abstract

Aims/hypothesis

Type 1 diabetes results from the autoimmune destruction of pancreatic beta cells. Exogenous insulin therapy cannot achieve precise physiological control of blood glucose concentrations, and debilitating complications develop. Lentiviral vectors are promising tools for liver-directed gene therapy. However, to date, transduction rates in vivo remain low in hepatocytes, without the induction of cell cycling. We investigated long-term transgene expression in quiescent hepatocytes in vitro and determined whether the lentiviral delivery of furin-cleavable insulin to the liver could reverse diabetes in rats.

Materials and methods

To improve transduction efficiency in vitro, we optimised hepatocyte isolation and maintenance protocols and, using an improved surgical delivery method, delivered furin-cleavable insulin alone or empty vector to the livers of streptozotocin-induced diabetic rats by means of a lentiviral vector. Rats were monitored for changes in body weight and blood glucose, and intravenous glucose tolerance tests were performed. Expression of insulin was determined by RT-PCR, immunohistochemistry and electron microscopy.

Results

We achieved long-term transgene expression in quiescent hepatocytes in vitro (87 ± 1.2% transduction efficiency), with up to 60 ± 3.2% transduction in vivo. We normalised blood glucose for 500 days—a significantly longer period than previously reported—making this the first successful study using a lentiviral vector. This procedure resulted in the expression of genes encoding several beta cell transcription factors, some pancreatic endocrine transdifferentiation, hepatic insulin storage in granules, and restoration of glucose tolerance. Liver function tests remained normal. Importantly, pancreatic exocrine transdifferentiation did not occur.

Conclusions/interpretation

Our data suggest that this regimen may ultimately be employed for the treatment of type 1 diabetes.
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Metadata
Title
Long-term correction of diabetes in rats after lentiviral hepatic insulin gene therapy
Authors
B. Ren
B. A. O’Brien
M. A. Swan
M. E. Koina
N. Nassif
M. Q. Wei
A. M. Simpson
Publication date
01-09-2007
Publisher
Springer-Verlag
Published in
Diabetologia / Issue 9/2007
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-007-0722-0

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