title: Reprogramming Mouse Cells With a Pancreatic Duct Phenotype to Insulin-Producing β-Like Cells.
abstract: Reprogramming technology has opened the possibility of converting one cell type into another by forced expression of transgenes. Transduction of adenoviral vectors encoding 3 pancreatic transcription factors, Pdx1, Ngn3, and MafA, into mouse pancreas results in direct reprogramming of exocrine cells to insulin-producing β-like cells. We hypothesized that cultured adult pancreatic duct cells could be reprogrammed to become insulin-producing β-cells by adenoviral-mediated expression of this same combination of factors. Exocrine were isolated from adult mouse insulin 1 promoter (MIP)-green fluorescent protein (GFP) transgenic mice to allow new insulin-expressing cells to be detected by GFP fluorescence. Cultured cells were transduced by an adenoviral vector carrying a polycistronic construct Ngn3/Pdx1/MafA/mCherry (Ad-M3C) or mCherry sequence alone as a control vector. In addition, the effects of glucagon-like peptide-1 (GLP-1) receptor agonist, exendin-4 (Ex-4) on the reprogramming process were examined. GFP(+) cells appeared 2 days after Ad-M3C transduction; the reprogramming efficiency was 8.6 ± 2.6% by day 4 after transduction. Ad-M3C also resulted in increased expression of β-cell markers insulin 1 and 2, with enhancement by Ex-4. Expression of other β-cell markers, neuroD and GLP-1 receptor, were also significantly up-regulated. The amount of insulin release into the media and insulin content of the cells were significantly higher in the Ad-M3C-transduced cells; this too was enhanced by Ex-4. The transduced cells did not secrete insulin in response to increased glucose, indicating incomplete differentiation to β-cells. Thus, cultured murine adult pancreatic cells with a duct phenotype can be directly reprogrammed to insulin-producing β-like cells by adenoviral delivery of 3 pancreatic transcription factors.
authors.affiliation: Section on Islet Cell and Regenerative Biology (T.Y., C.C.-W., F.C., P.A.L., L.G., A.S., S.B.-W., G.C.W.), Research Division, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts 02215; and Department of Stem Cell and Regenerative Biology (W.L., Q.Z.), Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts 02138.
authors.forename: Takatsugu
authors.initials: T
authors.lastname: Yamada
authors.forename: Claudia
authors.initials: C
authors.lastname: Cavelti-Weder
authors.forename: Francisco
authors.initials: F
authors.lastname: Caballero
authors.forename: Philippe A
authors.initials: PA
authors.lastname: Lysy
authors.forename: Lili
authors.initials: L
authors.lastname: Guo
authors.forename: Arun
authors.initials: A
authors.lastname: Sharma
authors.forename: Weida
authors.initials: W
authors.lastname: Li
authors.forename: Qiao
authors.initials: Q
authors.lastname: Zhou
authors.forename: Susan
authors.initials: S
authors.lastname: Bonner-Weir
authors.forename: Gordon C
authors.initials: GC
authors.lastname: Weir
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology*; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010183:Pancreatic Ducts / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D014157:Transcription Factors
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