Pancreatic transdifferentiation in porcine liver following lentiviral delivery of human furin-cleavable insulin

Gerace, D; Ren, B; Hawthorne, WJ; Byrne, MR; Phillips, PM; O'Brien, BA; Nassif, N; Alexander, IE; Simpson, AM

Pancreatic transdifferentiation in porcine liver following lentiviral delivery of human furin-cleavable insulin

Gerace, D

Ren, B Hawthorne, WJ Byrne, MR Phillips, PM O'Brien, BA

Nassif, N

Alexander, IE Simpson, AM

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Publication Type:: Journal Article
Citation:: Transplantation Proceedings, 2013, 45 (5), pp. 1869 - 1874
Issue Date:: 2013-06-01

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Field	Value	Language
dc.contributor.author	Gerace, D https://orcid.org/0000-0001-9154-0624	en_US
dc.contributor.author	Ren, B	en_US
dc.contributor.author	Hawthorne, WJ	en_US
dc.contributor.author	Byrne, MR	en_US
dc.contributor.author	Phillips, PM	en_US
dc.contributor.author	O'Brien, BA https://orcid.org/0000-0001-5887-2424	en_US
dc.contributor.author	Nassif, N https://orcid.org/0000-0003-2675-8322	en_US
dc.contributor.author	Alexander, IE	en_US
dc.contributor.author	Simpson, AM https://orcid.org/0000-0002-2249-5097	en_US
dc.date.available	2013-01-03	en_US
dc.date.issued	2013-06-01	en_US
dc.identifier.citation	Transplantation Proceedings, 2013, 45 (5), pp. 1869 - 1874	en_US
dc.identifier.issn	0041-1345	en_US
dc.identifier.uri	http://hdl.handle.net/10453/26837
dc.description.abstract	Type I diabetes mellitus (TID) results from the autoimmune destruction of the insulin-producing pancreatic β-cells. Gene therapy is one strategy being actively explored to cure TID by affording non-β-cells the ability to secrete insulin in response to physiologic stimuli. In previous studies, we used a novel surgical technique to express furin-cleavable human insulin (INS-FUR) in the livers of streptozotocin (STZ)-diabetic Wistar rats and nonobese diabetic (NOD) mice with the use of the HMD lentiviral vector. Normoglycemia was observed for 500 and 150 days, respectively (experimental end points). Additionally, some endocrine transdifferentiation of the liver, with storage of insulin in granules, and expression of some β-cell transcription factors (eg, Pdx1, Neurod1, Neurog3, Nkx2-2, Pax4) and pancreatic hormones in both studies. The aim of this study was to determine if this novel approach could induce liver to pancreatic transdifferentiation to reverse diabetes in pancreatectomized Westran pigs. Nine pigs were used in the study, however only one pig maintained normal fasting blood glucose levels for the period from 10 to 44 days (experimental end point). This animal was given 2.8 × 10 9 transducing units/kg of the lentiviral vector expressing INS-FUR. A normal intravenous glucose tolerance test was achieved at 30 days. Reverse-transcription polymerase chain reaction analysis of the liver tissue revealed expression of several β-cell transcription factors, including the key factors, Pdx-1 and Neurod1, pancreatic hormones, glucagon, and somatostatin; however, endogenous pig insulin was not expressed. Triple immunofluorescence showed extensive insulin expression, as was previously observed in our studies with rodents. Additionally, a small amount of glucagon and somatostatin protein expression was seen. Collectively, these data indicate that pancreatic transdifferentiation of the liver tissue had occurred. Our data suggest that this regimen may ultimately be used clinically to cure TID, however more work is required to replicate the successful reversal of diabetes in increased numbers of pigs. © 2013 Elsevier Inc.	en_US
dc.relation	http://purl.org/au-research/grants/nhmrc/513100
dc.relation.ispartof	Transplantation Proceedings	en_US
dc.relation.isbasedon	10.1016/j.transproceed.2013.01.051	en_US
dc.subject.mesh	Liver	en_US
dc.subject.mesh	Pancreas	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Swine	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Lentivirus	en_US
dc.subject.mesh	Insulin	en_US
dc.subject.mesh	Furin	en_US
dc.subject.mesh	Polymerase Chain Reaction	en_US
dc.subject.mesh	Cell Differentiation	en_US
dc.title	Pancreatic transdifferentiation in porcine liver following lentiviral delivery of human furin-cleavable insulin	en_US
dc.type	Journal Article
utslib.citation.volume	5	en_US
utslib.citation.volume	45	en_US
utslib.for	1103 Clinical Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
dc.location.activity	Robert Koch Inst, Berlin, GERMANY
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	5	en_US
pubs.publication-status	Published	en_US
pubs.volume	45	en_US

Abstract:

Type I diabetes mellitus (TID) results from the autoimmune destruction of the insulin-producing pancreatic β-cells. Gene therapy is one strategy being actively explored to cure TID by affording non-β-cells the ability to secrete insulin in response to physiologic stimuli. In previous studies, we used a novel surgical technique to express furin-cleavable human insulin (INS-FUR) in the livers of streptozotocin (STZ)-diabetic Wistar rats and nonobese diabetic (NOD) mice with the use of the HMD lentiviral vector. Normoglycemia was observed for 500 and 150 days, respectively (experimental end points). Additionally, some endocrine transdifferentiation of the liver, with storage of insulin in granules, and expression of some β-cell transcription factors (eg, Pdx1, Neurod1, Neurog3, Nkx2-2, Pax4) and pancreatic hormones in both studies. The aim of this study was to determine if this novel approach could induce liver to pancreatic transdifferentiation to reverse diabetes in pancreatectomized Westran pigs. Nine pigs were used in the study, however only one pig maintained normal fasting blood glucose levels for the period from 10 to 44 days (experimental end point). This animal was given 2.8 × 10 9 transducing units/kg of the lentiviral vector expressing INS-FUR. A normal intravenous glucose tolerance test was achieved at 30 days. Reverse-transcription polymerase chain reaction analysis of the liver tissue revealed expression of several β-cell transcription factors, including the key factors, Pdx-1 and Neurod1, pancreatic hormones, glucagon, and somatostatin; however, endogenous pig insulin was not expressed. Triple immunofluorescence showed extensive insulin expression, as was previously observed in our studies with rodents. Additionally, a small amount of glucagon and somatostatin protein expression was seen. Collectively, these data indicate that pancreatic transdifferentiation of the liver tissue had occurred. Our data suggest that this regimen may ultimately be used clinically to cure TID, however more work is required to replicate the successful reversal of diabetes in increased numbers of pigs. © 2013 Elsevier Inc.

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