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Ectopic expression of neurogenin 3 in neonatal pig pancreatic precursor cells induces (trans)differentiation to functional alpha cells

Authors
  • Harb, G.1
  • Heremans, Y.2
  • Heimberg, H.2
  • Korbutt, G. S.1
  • 1 University of Alberta, Department of Surgery, Surgical-Medical Research Institute, 1074 Dentistry Pharmacy Center, Edmonton, AB, T6G 2N8, Canada , Edmonton (Canada)
  • 2 Brussels Free University-VUB, Diabetes Research Center, Brussels, Belgium , Brussels (Belgium)
Type
Published Article
Journal
Diabetologia
Publisher
Springer-Verlag
Publication Date
May 31, 2006
Volume
49
Issue
8
Pages
1855–1863
Identifiers
DOI: 10.1007/s00125-006-0299-z
Source
Springer Nature
Keywords
License
Yellow

Abstract

Aims/hypothesisNeurogenin 3 (NEUROG3), a basic helix-loop-helix transcription factor that is needed for endocrine cell development in the embryonic pancreas, has been shown to induce transdifferentiation of duct cells from adult pancreas towards a neuro-endocrine phenotype. Our study explored the endocrine transdifferentiation potential of NEUROG3 in neonatal pancreatic precursor cells.Materials and methodsA replication-deficient adenovirus expressing Neurog3 and green fluorescent protein (GFP) (Ad-NEUROG3) was used to infect neonatal pig pancreatic cell preparations enriched for endocrine islet and cytokeratin-positive precursor cells. GFP-positive cells were sorted using flow cytometry on days 3 and 8 after infection and characterised at the transcript and protein level. For in vivo experiments, the total population of Ad-NEUROG3-infected pancreatic cells was transplanted, then later removed for determination of graft hormone content and immunohistochemistry.ResultsAmong the GFP-positive cells, the fraction of precursor cells decreased by more than 85% at day 8 after infection, while the fraction of glucagon-positive cells increased 2.5-fold and the beta cell number remained the same. Transplantation of the Ad-NEUROG3-infected pancreatic cell preparation failed to reverse streptozotocin-induced hyperglycaemia, while non-infected cells and a control cell preparation infected with replication-deficient adenovirus expressing only GFP were able to do so. At day 109 after transplantation, kidneys grafted with Ad-NEUROG3-infected pancreatic cells contained significantly decreased insulin and increased glucagon levels. Abundant glucagon-immunopositive cells were seen in Ad-NEUROG3-infected grafts, which were virtually devoid of proliferating insulin-positive cells.Conclusions/interpretationIn summary, adenoviral delivery of NEUROG3 to pancreatic precursor cells from neonatal pig pancreas promotes alpha cell differentiation in vitro and in vivo.

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