In vitro reprogramming of pancreatic alpha cells towards a beta cell phenotype following ectopic HNF4 alpha, expression

Autors de IIS La Fe

• 

  Ramiro Jover Atienza

  Autor

Autors aliens a IIS La Fe

• Sangan CB
• Heimberg H
• Tosh D

Grups d'Investigació

• Hepatología experimental y Trasplante hepático

  Investigador Principal

  Laia Tolosa Pardo

Abstract

There is currently a shortage of organ donors available for pancreatic beta cell transplantation into diabetic patients. An alternative source of beta cells is pre-existing pancreatic cells. While we know that beta cells can arise directly from alpha cells during pancreatic regeneration we do not understand the molecular basis for the switch in phenotype. The aim of the present study was to investigate if hepatocyte nuclear factor 4 alpha (HNF4 alpha), a transcription factor essential for a normal beta cell phenotype, could induce the reprogramming of alpha cells towards potential beta cells. We utilised an in vitro model of pancreatic alpha cells, the murine alpha TC1-9 cell line. We initially characterised the aTC1-9 cell line before and following adenovirus-mediated ectopic expression of HNF4 alpha. We analysed the phenotype at transcript and protein level and assessed its glucose-responsiveness. Ectopic HNF4 alpha expression in the aTC1-9 cell line induced a change in morphology (17-fold increase in size), suppressed glucagon expression, induced key beta cell-specific markers (insulin, C-peptide, glucokinase, GLUT2 and Pax4) and pancreatic polypeptide (PP) and enabled the cells to secrete insulin in a glucose-regulated manner. In conclusion, HNF4 alpha reprograms alpha cells to beta-like cells. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Keywords

• HNF4 alpha; Alpha cell; Beta cell; Insulin; Glucagon; Pancreatic polypeptide