Reprogramming Mouse Cells With a Pancreatic Duct Phenotype to Insulin-Producing β-Like Cells.

TitleReprogramming Mouse Cells With a Pancreatic Duct Phenotype to Insulin-Producing β-Like Cells.
Publication TypeJournal Article
Year of Publication2015
AuthorsYamada T, Cavelti-Weder C, Caballero F, Lysy PA, Guo L, Sharma A, Li W, Zhou Q, Bonner-Weir S, Weir GC
Date Published2015 Jun
KeywordsAnimals, Cells, Cultured, Cellular Reprogramming, Insulin-Secreting Cells, Male, Mice, Mice, Transgenic, Pancreatic Ducts, Reverse Transcriptase Polymerase Chain Reaction, Transcription Factors, Transcriptome

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.

Alternate JournalEndocrinology
PubMed ID25836667
PubMed Central IDPMC4430605
Grant ListR01 DK093909 / DK / NIDDK NIH HHS / United States
P30 DK036836 / DK / NIDDK NIH HHS / United States
DK093909 / DK / NIDDK NIH HHS / United States
R01 DK066056 / DK / NIDDK NIH HHS / United States
R01DK066056 / DK / NIDDK NIH HHS / United States