title: Pancreatic Transdifferentiation and Glucose-Regulated Production of Human Insulin in the H4IIE Rat Liver Cell Line.
abstract: Due to the limitations of current treatment regimes, gene therapy is a promising strategy being explored to correct blood glucose concentrations in diabetic patients. In the current study, we used a retroviral vector to deliver either the human insulin gene alone, the rat NeuroD1 gene alone, or the human insulin gene and rat NeuroD1 genes together, to the rat liver cell line, H4IIE, to determine if storage of insulin and pancreatic transdifferentiation occurred. Stable clones were selected and expanded into cell lines: H4IIEins (insulin gene alone), H4IIE/ND (NeuroD1 gene alone), and H4IIEins/ND (insulin and NeuroD1 genes). The H4IIEins cells did not store insulin; however, H4IIE/ND and H4IIEins/ND cells stored 65.5 ± 5.6 and 1475.4 ± 171.8 pmol/insulin/5 × 10⁶ cells, respectively. Additionally, several β cell transcription factors and pancreatic hormones were expressed in both H4IIE/ND and H4IIEins/ND cells. Electron microscopy revealed insulin storage vesicles in the H4IIE/ND and H4IIEins/ND cell lines. Regulated secretion of insulin to glucose (0-20 mmol/L) was seen in the H4IIEins/ND cell line. The H4IIEins/ND cells were transplanted into diabetic immunoincompetent mice, resulting in normalization of blood glucose. This data shows that the expression of NeuroD1 and insulin in liver cells may be a useful strategy for inducing islet neogenesis and reversing diabetes.
authors.affiliation: School of Life Sciences and Centre for Health Technologies, University of Technology Sydney, P.O. Box 123, Broadway, 2007 Sydney, NSW, Australia. Binhai.Ren@uts.edu.au.
authors.forename: Binhai
authors.initials: B
authors.lastname: Ren
authors.affiliation: School of Life Sciences and Centre for Health Technologies, University of Technology Sydney, P.O. Box 123, Broadway, 2007 Sydney, NSW, Australia. Chang.Tao@sswahs.nsw.gov.au.
authors.forename: Chang
authors.initials: C
authors.lastname: Tao
authors.affiliation: School of Medical Sciences (Anatomy & Histology) and Bosch Institute, University of Sydney, 2006 Sydney, NSW, Australia. swan@anatomy.usyd.edu.au.
authors.forename: Margaret Anne
authors.initials: MA
authors.lastname: Swan
authors.affiliation: School of Medical Sciences (Anatomy & Histology) and Bosch Institute, University of Sydney, 2006 Sydney, NSW, Australia. nichole.joachim@gmail.com.
authors.forename: Nichole
authors.initials: N
authors.lastname: Joachim
authors.affiliation: School of Life Sciences and Centre for Health Technologies, University of Technology Sydney, P.O. Box 123, Broadway, 2007 Sydney, NSW, Australia. Rosetta.Martiniello-Wilks@uts.edu.au.
authors.forename: Rosetta
authors.initials: R
authors.lastname: Martiniello-Wilks
authors.affiliation: School of Life Sciences and Centre for Health Technologies, University of Technology Sydney, P.O. Box 123, Broadway, 2007 Sydney, NSW, Australia. Najah.Nassif@uts.edu.au.
authors.forename: Najah T
authors.initials: NT
authors.lastname: Nassif
authors.affiliation: School of Life Sciences and Centre for Health Technologies, University of Technology Sydney, P.O. Box 123, Broadway, 2007 Sydney, NSW, Australia. Bronwyn.Obrien@uts.edu.au.
authors.forename: Bronwyn A
authors.initials: BA
authors.lastname: O'Brien
authors.affiliation: School of Life Sciences and Centre for Health Technologies, University of Technology Sydney, P.O. Box 123, Broadway, 2007 Sydney, NSW, Australia. Ann.Simpson@uts.edu.au.
authors.forename: Ann M
authors.initials: AM
authors.lastname: Simpson
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D060846:Cell Engineering / Q000379:methods*; D002460:Cell Line; D054338:Cell Transdifferentiation; D003920:Diabetes Mellitus / Q000235:genetics / Q000628:therapy; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000235:genetics* / Q000378:metabolism*; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D016513:Mice, SCID; D051381:Rats
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005947:Glucose; D007328:Insulin; C490555:Neurod1 protein, rat
keywords: H4IIE cells; diabetes; furin-cleavable human insulin; gene therapy; insulin storage; liver cells; regulated insulin secretion; secretory granules; β cell transcription factors