Suppression of epithelial-to-mesenchymal transitioning enhances ex vivo reprogramming of human exocrine pancreatic tissue toward functional insulin-producing β-like cells

Maria Joao Lima, Kenneth R. Muir, Hilary M. Docherty, Robert Drummond, Neil W.A. McGowan, Yves Heremans, Shareen Forbes, Isabelle Houbracken, James A. Ross, Stuart J. Forbes, Philippe Ravassard, Harry Heimberg, John Casey, Kevin Docherty

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Abstract

Because of the lack of tissue available for islet transplantation, new sources of β-cells have been sought for the treatment of type 1 diabetes. The aim of this study was to determine whether the human exocrine-enriched fraction from the islet isolation procedure could be reprogrammed to provide additional islet tissue for transplantation. The exocrine-enriched cells rapidly dedifferentiated in culture and grew as a mesenchymal monolayer. Genetic lineage tracing confirmed that these mesenchymal cells arose, in part, through a process of epithelial-to-mesenchymal transitioning (EMT). A protocol was developed whereby transduction of these mesenchymal cells with adenoviruses containing Pdx1, Ngn3, MafA, and Pax4 generated a population of cells that were enriched in glucagon-secreting α-like cells. Transdifferentiation or reprogramming toward insulin-secreting β-cells was enhanced, however, when using unpassaged cells in combination with inhibition of EMT by inclusion of Rho-associated kinase (ROCK) and transforming growth factor-β1 inhibitors. Resultant cells were able to secrete insulin in response to glucose and on transplantation were able to normalize blood glucose levels in streptozotocin diabetic NOD/SCID mice. In conclusion, reprogramming of human exocrine-enriched tissue can be best achieved using fresh material under conditions whereby EMT is inhibited, rather than allowing the culture to expand as a mesenchymal monolayer.
Original languageEnglish
Pages (from-to) 2821-2833
Number of pages13
JournalDiabetes
Volume62
Issue number8
Early online date22 Apr 2013
DOIs
Publication statusPublished - Aug 2013

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Keywords

  • islet transplantation
  • stem cells
  • pancreatic transcription factors
  • gene therapy

Cite this

Lima, M. J., Muir, K. R., Docherty, H. M., Drummond, R., McGowan, N. W. A., Heremans, Y., Forbes, S., Houbracken, I., Ross, J. A., Forbes, S. J., Ravassard, P., Heimberg, H., Casey, J., & Docherty, K. (2013). Suppression of epithelial-to-mesenchymal transitioning enhances ex vivo reprogramming of human exocrine pancreatic tissue toward functional insulin-producing β-like cells. Diabetes, 62(8), 2821-2833 . https://doi.org/10.2337/db12-1256