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Sci Rep. 2018 Oct 18;8(1):15374. doi: 10.1038/s41598-018-33838-5.

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

Author information

1
Department of Oncology, University of Cambridge, Hutchison/MRC Research Centre, Hills Road, Cambridge, CB2 0XZ, UK. ra461@cam.ac.uk.
2
Wellcome - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK. ra461@cam.ac.uk.
3
Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge, CB3 0HE, UK. ra461@cam.ac.uk.
4
Max Planck Institute for the Physics of Complex Systems, Nöthnitzer Str. 38, 01187, Dresden, Germany.
5
Center for Systems Biology Dresden, Pfotenhauer Str. 108, 01307, Dresden, Germany.
6
Wellcome - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK.
7
Department of Haematology, Cambridge Institute for Medical Research, Hills Road, Cambridge, CB2 0XY, UK.
8
Department of Oncology, University of Cambridge, Hutchison/MRC Research Centre, Hills Road, Cambridge, CB2 0XZ, UK.
9
The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
10
UCL Great Ormond Street Institute of Child Health, 30 Guilford Street, London, WC1N 1EH, UK.
11
The Wellcome/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK.
12
Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge, CB3 0HE, UK.
13
Department of Oncology, University of Cambridge, Hutchison/MRC Research Centre, Hills Road, Cambridge, CB2 0XZ, UK. ap113@cam.ac.uk.
14
Wellcome - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK. ap113@cam.ac.uk.

Abstract

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.

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