Generation of Functional Beta-Like Cells from Human Exocrine Pancreas

Maria J. Lima, Kenneth R. Muir, Hilary M. Docherty, Neil W. A. McGowan, Shareen Forbes, Yves Heremans, Harry Heimberg, John Casey, Kevin Docherty

Research output: Contribution to journalArticle

7 Citations (Scopus)
4 Downloads (Pure)

Abstract

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.

Original languageEnglish
Article numbere0156204
Pages (from-to)1-19
Number of pages19
JournalPloS ONE
Volume11
Issue number5
DOIs
Publication statusPublished - 31 May 2016

Fingerprint

Exocrine Pancreas
pancreas
Insulin
Transcription Factors
insulin
Tissue
Medical problems
transcription factors
cells
Glucose
Proinsulin
Islets of Langerhans Transplantation
Secretory Vesicles
Insulin-Secreting Cells
Laminin
Therapeutic Uses
proinsulin
Type 1 Diabetes Mellitus
Islets of Langerhans
Transplants

Cite this

Lima, M. J., Muir, K. R., Docherty, H. M., McGowan, N. W. A., Forbes, S., Heremans, Y., ... Docherty, K. (2016). Generation of Functional Beta-Like Cells from Human Exocrine Pancreas. PloS ONE, 11(5), 1-19. [e0156204]. https://doi.org/10.1371/journal.pone.0156204

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.

In: PloS ONE, Vol. 11, No. 5, e0156204, 31.05.2016, p. 1-19.

Research output: Contribution to journalArticle

Lima, MJ, Muir, KR, Docherty, HM, McGowan, NWA, Forbes, S, Heremans, Y, Heimberg, H, Casey, J & Docherty, K 2016, 'Generation of Functional Beta-Like Cells from Human Exocrine Pancreas' PloS ONE, vol. 11, no. 5, e0156204, pp. 1-19. https://doi.org/10.1371/journal.pone.0156204
Lima MJ, Muir KR, Docherty HM, McGowan NWA, Forbes S, Heremans Y et al. Generation of Functional Beta-Like Cells from Human Exocrine Pancreas. PloS ONE. 2016 May 31;11(5):1-19. e0156204. https://doi.org/10.1371/journal.pone.0156204
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. In: PloS ONE. 2016 ; Vol. 11, No. 5. pp. 1-19.
@article{0b81a76ec6e94e719c9f398fe87376ed,
title = "Generation of Functional Beta-Like Cells from Human Exocrine Pancreas",
abstract = "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.",
author = "Lima, {Maria J.} and Muir, {Kenneth R.} and Docherty, {Hilary M.} and McGowan, {Neil W. A.} and Shareen Forbes and Yves Heremans and Harry Heimberg and John Casey and Kevin Docherty",
note = "Funding: This work was supported by a grant from the Medical Research Council MR/J015277/1. The Scottish National Islet Transplant Programme is funded by the National Services Division of NHS Scotland. KRM was funded by a Fellowship from the Wellcome Trust / Scottish Translational Medicine and Therapeutics Initiative 85664. Acknowledgments This work was supported by a grant from the Medical Research Council MR/J015277/1. The Scottish National Islet Transplant Programme is funded by the National Services Division of NHS Scotland. KRM was funded by a Fellowship from the Wellcome Trust/ Scottish Translational Medicine and Therapeutics Initiative 85664. We thank Joanna Sweetman for assistance in optimisation of the immunogold staining.",
year = "2016",
month = "5",
day = "31",
doi = "10.1371/journal.pone.0156204",
language = "English",
volume = "11",
pages = "1--19",
journal = "PloS ONE",
issn = "1932-6203",
publisher = "PUBLIC LIBRARY SCIENCE",
number = "5",

}

TY - JOUR

T1 - Generation of Functional Beta-Like Cells from Human Exocrine Pancreas

AU - Lima, Maria J.

AU - Muir, Kenneth R.

AU - Docherty, Hilary M.

AU - McGowan, Neil W. A.

AU - Forbes, Shareen

AU - Heremans, Yves

AU - Heimberg, Harry

AU - Casey, John

AU - Docherty, Kevin

N1 - Funding: This work was supported by a grant from the Medical Research Council MR/J015277/1. The Scottish National Islet Transplant Programme is funded by the National Services Division of NHS Scotland. KRM was funded by a Fellowship from the Wellcome Trust / Scottish Translational Medicine and Therapeutics Initiative 85664. Acknowledgments This work was supported by a grant from the Medical Research Council MR/J015277/1. The Scottish National Islet Transplant Programme is funded by the National Services Division of NHS Scotland. KRM was funded by a Fellowship from the Wellcome Trust/ Scottish Translational Medicine and Therapeutics Initiative 85664. We thank Joanna Sweetman for assistance in optimisation of the immunogold staining.

PY - 2016/5/31

Y1 - 2016/5/31

N2 - 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.

AB - 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.

U2 - 10.1371/journal.pone.0156204

DO - 10.1371/journal.pone.0156204

M3 - Article

VL - 11

SP - 1

EP - 19

JO - PloS ONE

JF - PloS ONE

SN - 1932-6203

IS - 5

M1 - e0156204

ER -