Neurogenin3 phosphorylation controls reprogramming efficiency of pancreatic ductal organoids into endocrine cells.

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Nestorowa, Sonia 
Davies, John 
Campinoti, Sara 

β-cell replacement has been proposed as an effective treatment for some forms of diabetes, and in vitro methods for β-cell generation are being extensively explored. A potential source of β-cells comes from fate conversion of exocrine pancreatic cells into the endocrine lineage, by overexpression of three regulators of pancreatic endocrine formation and β-cell identity, Ngn3, Pdx1 and MafA. Pancreatic ductal organoid cultures have recently been developed that can be expanded indefinitely, while maintaining the potential to differentiate into the endocrine lineage. Here, using mouse pancreatic ductal organoids, we see that co-expression of Ngn3, Pdx1 and MafA are required and sufficient to generate cells that express insulin and resemble β-cells transcriptome-wide. Efficiency of β-like cell generation can be significantly enhanced by preventing phosphorylation of Ngn3 protein and further augmented by conditions promoting differentiation. Taken together, our new findings underline the potential of ductal organoid cultures as a source material for generation of β-like cells and demonstrate that post-translational regulation of reprogramming factors can be exploited to enhance β-cell generation.

Animals, Basic Helix-Loop-Helix Transcription Factors, Cellular Reprogramming, HEK293 Cells, Humans, Insulin-Secreting Cells, Mice, Nerve Tissue Proteins, Organoids, Pancreatic Ducts, Phosphorylation
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Springer Science and Business Media LLC
Wellcome Trust (098357/Z/12/Z)
Leukaemia & Lymphoma Research (12029)
Wellcome Trust (097922/Z/11/Z)
Medical Research Council (MC_PC_12009)
Medical Research Council (MR/K018329/1)
Medical Research Council (MR/L021129/1)
Medical Research Council (MR/M008975/1)
This work was supported by: the MRC Research Grant MR/K018329/1 and the Rosetrees Trust and Stoneygate Trust (to A.P. and R.A.); the MRC Research Grant MR/L021129/1 and core support from the Wellcome Trust and MRC Cambridge Stem Cell Institute (to A.P., R.A., B.D.S., B.G.); the Wellcome Trust 098357/Z/12/Z (to B.D.S. and R.A.); the Wellcome Trust 097922/Z/11/Z and the Clinical Research Infrastructure Single-cell Facility MR/M008975/1 (to B.G.). S.C. is supported by a GOSH Charity studentship (V6116). P.B. is supported by the UCL Excellence Fellowship Programme, the European Research Council (ERC-Stg-2014 639429), the Rosetrees Trust (A1411 and A1179) and the NIHR BRC at Great Ormond Street Hospital for Children NHS Foundation Trust. M.H. is a Sir Henry Dale fellow and supported by an EU-H2020 grant, LSMF4LIFE, by the Wellcome Trust 104151/Z/14/A and the Royal Society