Generation of Functional Beta-Like Cells from Human Exocrine Pancreas

Transcription factor mediated lineage reprogramming of human pancreatic exocrine tissue could conceivably provide an unlimited supply of islets for transplantation in the treatment of diabetes. Exocrine tissue can be efficiently reprogrammed to islet-like cells using a cocktail of transcription fact...

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Main Authors:	Lima, Maria J., Muir, Kenneth R., Docherty, Hilary M., McGowan, Neil W. A., Forbes, Shareen, Heremans, Yves, Heimberg, Harry, Casey, John, Docherty, Kevin
Format:	Online
Language:	English
Published:	Public Library of Science 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4887015/

id	pubmed-4887015
recordtype	oai_dc
spelling	pubmed-48870152016-06-10 Generation of Functional Beta-Like Cells from Human Exocrine Pancreas Lima, Maria J. Muir, Kenneth R. Docherty, Hilary M. McGowan, Neil W. A. Forbes, Shareen Heremans, Yves Heimberg, Harry Casey, John Docherty, Kevin Research Article Transcription factor mediated lineage reprogramming of human pancreatic exocrine tissue could conceivably provide an unlimited supply of islets for transplantation in the treatment of diabetes. Exocrine tissue can be efficiently reprogrammed to islet-like cells using a cocktail of transcription factors: Pdx1, Ngn3, MafA and Pax4 in combination with growth factors. We show here that overexpression of exogenous Pax4 in combination with suppression of the endogenous transcription factor ARX considerably enhances the production of functional insulin-secreting β-like cells with concomitant suppression of α-cells. The efficiency was further increased by culture on laminin-coated plates in media containing low glucose concentrations. Immunocytochemistry revealed that reprogrammed cultures were composed of ~45% islet-like clusters comprising >80% monohormonal insulin+ cells. The resultant β-like cells expressed insulin protein levels at ~15–30% of that in adult human islets, efficiently processed proinsulin and packaged insulin into secretory granules, exhibited glucose responsive insulin secretion, and had an immediate and prolonged effect in normalising blood glucose levels upon transplantation into diabetic mice. We estimate that approximately 3 billion of these cells would have an immediate therapeutic effect following engraftment in type 1 diabetes patients and that one pancreas would provide sufficient tissue for numerous transplants. Public Library of Science 2016-05-31 /pmc/articles/PMC4887015/ /pubmed/27243814 http://dx.doi.org/10.1371/journal.pone.0156204 Text en © 2016 Lima et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Lima, Maria J. Muir, Kenneth R. Docherty, Hilary M. McGowan, Neil W. A. Forbes, Shareen Heremans, Yves Heimberg, Harry Casey, John Docherty, Kevin
spellingShingle	Lima, Maria J. Muir, Kenneth R. Docherty, Hilary M. McGowan, Neil W. A. Forbes, Shareen Heremans, Yves Heimberg, Harry Casey, John Docherty, Kevin Generation of Functional Beta-Like Cells from Human Exocrine Pancreas
author_facet	Lima, Maria J. Muir, Kenneth R. Docherty, Hilary M. McGowan, Neil W. A. Forbes, Shareen Heremans, Yves Heimberg, Harry Casey, John Docherty, Kevin
author_sort	Lima, Maria J.
title	Generation of Functional Beta-Like Cells from Human Exocrine Pancreas
title_short	Generation of Functional Beta-Like Cells from Human Exocrine Pancreas
title_full	Generation of Functional Beta-Like Cells from Human Exocrine Pancreas
title_fullStr	Generation of Functional Beta-Like Cells from Human Exocrine Pancreas
title_full_unstemmed	Generation of Functional Beta-Like Cells from Human Exocrine Pancreas
title_sort	generation of functional beta-like cells from human exocrine pancreas
description	Transcription factor mediated lineage reprogramming of human pancreatic exocrine tissue could conceivably provide an unlimited supply of islets for transplantation in the treatment of diabetes. Exocrine tissue can be efficiently reprogrammed to islet-like cells using a cocktail of transcription factors: Pdx1, Ngn3, MafA and Pax4 in combination with growth factors. We show here that overexpression of exogenous Pax4 in combination with suppression of the endogenous transcription factor ARX considerably enhances the production of functional insulin-secreting β-like cells with concomitant suppression of α-cells. The efficiency was further increased by culture on laminin-coated plates in media containing low glucose concentrations. Immunocytochemistry revealed that reprogrammed cultures were composed of ~45% islet-like clusters comprising >80% monohormonal insulin+ cells. The resultant β-like cells expressed insulin protein levels at ~15–30% of that in adult human islets, efficiently processed proinsulin and packaged insulin into secretory granules, exhibited glucose responsive insulin secretion, and had an immediate and prolonged effect in normalising blood glucose levels upon transplantation into diabetic mice. We estimate that approximately 3 billion of these cells would have an immediate therapeutic effect following engraftment in type 1 diabetes patients and that one pancreas would provide sufficient tissue for numerous transplants.
publisher	Public Library of Science
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4887015/
_version_	1613586605849706496

Generation of Functional Beta-Like Cells from Human Exocrine Pancreas

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