Long-term reversal of diabetes in non-obese diabetic mice by liver-directed gene therapy

Ren, B; O'Brien, BA; Byrne, MR; Ch'ng, E; Gatt, PN; Swan, MA; Nassif, NT; Wei, MQ; Gijsbers, R; Debyser, Z; Simpson, AM

Long-term reversal of diabetes in non-obese diabetic mice by liver-directed gene therapy

Ren, B O'Brien, BA

Byrne, MR Ch'ng, E Gatt, PN Swan, MA Nassif, NT

Wei, MQ Gijsbers, R Debyser, Z Simpson, AM

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Publication Type:: Journal Article
Citation:: Journal of Gene Medicine, 2013, 15 (1), pp. 28 - 41
Issue Date:: 2013-01-01

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Field	Value	Language
dc.contributor.author	Ren, B	en_US
dc.contributor.author	O'Brien, BA https://orcid.org/0000-0001-5887-2424	en_US
dc.contributor.author	Byrne, MR	en_US
dc.contributor.author	Ch'ng, E	en_US
dc.contributor.author	Gatt, PN	en_US
dc.contributor.author	Swan, MA	en_US
dc.contributor.author	Nassif, NT https://orcid.org/0000-0003-2675-8322	en_US
dc.contributor.author	Wei, MQ	en_US
dc.contributor.author	Gijsbers, R	en_US
dc.contributor.author	Debyser, Z	en_US
dc.contributor.author	Simpson, AM https://orcid.org/0000-0002-2249-5097	en_US
dc.date.available	2012-12-20	en_US
dc.date.issued	2013-01-01	en_US
dc.identifier.citation	Journal of Gene Medicine, 2013, 15 (1), pp. 28 - 41	en_US
dc.identifier.issn	1099-498X	en_US
dc.identifier.uri	http://hdl.handle.net/10453/27654
dc.description.abstract	Background: Type 1 diabetes (T1D) results from an autoimmune attack against the insulin-producing β-cells of the pancreas. The present study aimed to reverse T1D by gene therapy. Methods: We used a novel surgical technique, which involves isolating the liver from the circulation before the delivery of a lentiviral vector carrying furin-cleavable human insulin (INS-FUR) or empty vector to the livers of diabetic non-obese diabetic mice (NOD). This was compared with the direct injection of the vector into the portal circulation. Mice were monitored for body weight and blood glucose. Intravenous glucose tolerance tests were performed. Expression of insulin and pancreatic transcription factors was determined by the reverse transcriptase-polymerase chain reaction and immunohistochemistry and immunoelectron microscopy was used to localise insulin. Results: Using the novel surgical technique, we achieved long-term transduction (42% efficiency) of hepatocytes, restored normoglycaemia for 150 days (experimental endpoint) and re-established normal glucose tolerance. We showed the expression of β-cell transcription factors, murine insulin, glucagon and somatostatin, and hepatic storage of insulin in granules. The expression of hepatic markers, C/EBP-β, G6PC, AAT and GLUI was down-regulated in INS-FUR-treated livers. Liver function tests remained normal, with no evidence of intrahepatic inflammation or autoimmune destruction of the insulin-secreting liver tissue. By comparison, direct injection of INS-FUR reduced blood glucose levels, and no pancreatic transdifferentiation or normal glucose tolerance was observed. Conclusions: This gene therapy protocol has, for the first time, permanently reversed T1D with normal glucose tolerance in NOD mice and, as such, represents a novel therapeutic strategy for the treatment of T1D. © 2013 John Wiley & Sons, Ltd.	en_US
dc.relation	http://purl.org/au-research/grants/nhmrc/352909
dc.relation.ispartof	Journal of Gene Medicine	en_US
dc.relation.isbasedon	10.1002/jgm.2692	en_US
dc.subject.classification	Biotechnology	en_US
dc.subject.mesh	Liver	en_US
dc.subject.mesh	Hepatocytes	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Mice, Inbred NOD	en_US
dc.subject.mesh	Mice	en_US
dc.subject.mesh	Lentivirus	en_US
dc.subject.mesh	Diabetes Mellitus, Type 1	en_US
dc.subject.mesh	Somatostatin	en_US
dc.subject.mesh	Glucagon	en_US
dc.subject.mesh	Insulin	en_US
dc.subject.mesh	Furin	en_US
dc.subject.mesh	Alanine Transaminase	en_US
dc.subject.mesh	Aspartate Aminotransferases	en_US
dc.subject.mesh	Blood Glucose	en_US
dc.subject.mesh	Homeodomain Proteins	en_US
dc.subject.mesh	Trans-Activators	en_US
dc.subject.mesh	Nerve Tissue Proteins	en_US
dc.subject.mesh	Transcription Factors	en_US
dc.subject.mesh	Glucose Tolerance Test	en_US
dc.subject.mesh	Oligonucleotide Array Sequence Analysis	en_US
dc.subject.mesh	Transduction, Genetic	en_US
dc.subject.mesh	Genetic Vectors	en_US
dc.subject.mesh	Female	en_US
dc.subject.mesh	Insulin-Secreting Cells	en_US
dc.subject.mesh	Paired Box Transcription Factors	en_US
dc.subject.mesh	Basic Helix-Loop-Helix Transcription Factors	en_US
dc.subject.mesh	Cell Transdifferentiation	en_US
dc.subject.mesh	Genetic Therapy	en_US
dc.subject.mesh	Zebrafish Proteins	en_US
dc.title	Long-term reversal of diabetes in non-obese diabetic mice by liver-directed gene therapy	en_US
dc.type	Journal Article
utslib.citation.volume	1	en_US
utslib.citation.volume	15	en_US
utslib.for	1103 Clinical Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
utslib.copyright.status	closed_access
pubs.issue	1	en_US
pubs.publication-status	Published	en_US
pubs.volume	15	en_US

Abstract:

Background: Type 1 diabetes (T1D) results from an autoimmune attack against the insulin-producing β-cells of the pancreas. The present study aimed to reverse T1D by gene therapy. Methods: We used a novel surgical technique, which involves isolating the liver from the circulation before the delivery of a lentiviral vector carrying furin-cleavable human insulin (INS-FUR) or empty vector to the livers of diabetic non-obese diabetic mice (NOD). This was compared with the direct injection of the vector into the portal circulation. Mice were monitored for body weight and blood glucose. Intravenous glucose tolerance tests were performed. Expression of insulin and pancreatic transcription factors was determined by the reverse transcriptase-polymerase chain reaction and immunohistochemistry and immunoelectron microscopy was used to localise insulin. Results: Using the novel surgical technique, we achieved long-term transduction (42% efficiency) of hepatocytes, restored normoglycaemia for 150 days (experimental endpoint) and re-established normal glucose tolerance. We showed the expression of β-cell transcription factors, murine insulin, glucagon and somatostatin, and hepatic storage of insulin in granules. The expression of hepatic markers, C/EBP-β, G6PC, AAT and GLUI was down-regulated in INS-FUR-treated livers. Liver function tests remained normal, with no evidence of intrahepatic inflammation or autoimmune destruction of the insulin-secreting liver tissue. By comparison, direct injection of INS-FUR reduced blood glucose levels, and no pancreatic transdifferentiation or normal glucose tolerance was observed. Conclusions: This gene therapy protocol has, for the first time, permanently reversed T1D with normal glucose tolerance in NOD mice and, as such, represents a novel therapeutic strategy for the treatment of T1D. © 2013 John Wiley & Sons, Ltd.

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http://hdl.handle.net/10453/27654