Demethylation of induced pluripotent stem cells from type 1 diabetic patients enhances differentiation into functional pancreatic β cells

J Biol Chem. 2017 Aug 25;292(34):14066-14079. doi: 10.1074/jbc.M117.784280. Epub 2017 Mar 30.

Abstract

Type 1 diabetes (T1D) can be managed by transplanting either the whole pancreas or isolated pancreatic islets. However, cadaveric pancreas is scarcely available for clinical use, limiting this approach. As such, there is a great need to identify alternative sources of clinically usable pancreatic tissues. Here, we used induced pluripotent stem (iPS) cells derived from patients with T1D to generate glucose-responsive, insulin-producing cells (IPCs) via 3D culture. Initially, T1D iPS cells were resistant to differentiation, but transient demethylation treatment significantly enhanced IPC yield. The cells responded to high-glucose stimulation by secreting insulin in vitro The shape, size, and number of their granules, as observed by transmission electron microscopy, were identical to those found in cadaveric β cells. When the IPCs were transplanted into immunodeficient mice that had developed streptozotocin-induced diabetes, they promoted a dramatic decrease in hyperglycemia, causing the mice to become normoglycemic within 28 days. None of the mice died or developed teratomas. Because the cells are derived from "self," immunosuppression is not required, providing a much safer and reliable treatment option for T1D patients. Moreover, these cells can be used for drug screening, thereby accelerating drug discovery. In conclusion, our approach eliminates the need for cadaveric pancreatic tissue.

Keywords: 3D differentiation; beta cell; cell differentiation; diabetes; iPS cell; iPSC; induced pluripotent stem cell; insulin producing cells; insulin secretion; pancreatic islet; type 1 diabetes.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Azacitidine / analogs & derivatives
  • Azacitidine / pharmacology
  • Cadaver
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • DNA Methylation / drug effects*
  • DNA Modification Methylases / antagonists & inhibitors*
  • DNA Modification Methylases / metabolism
  • Decitabine
  • Diabetes Mellitus, Experimental / blood
  • Diabetes Mellitus, Experimental / immunology
  • Diabetes Mellitus, Experimental / surgery
  • Diabetes Mellitus, Type 1 / metabolism*
  • Diabetes Mellitus, Type 1 / pathology
  • Diabetes Mellitus, Type 1 / surgery
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Hyperglycemia / prevention & control
  • Induced Pluripotent Stem Cells / drug effects*
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / pathology
  • Induced Pluripotent Stem Cells / ultrastructure
  • Insulin / biosynthesis
  • Insulin / metabolism*
  • Insulin Secretion
  • Insulin-Secreting Cells / metabolism*
  • Insulin-Secreting Cells / transplantation
  • Insulin-Secreting Cells / ultrastructure
  • Mice, Knockout
  • Microscopy, Electron, Transmission
  • Organoids / metabolism*
  • Organoids / transplantation
  • Organoids / ultrastructure
  • Secretory Vesicles / metabolism
  • Secretory Vesicles / ultrastructure
  • Tissue Scaffolds
  • Transplantation, Heterologous / adverse effects
  • Transplantation, Heterotopic / adverse effects

Substances

  • Enzyme Inhibitors
  • Insulin
  • Decitabine
  • DNA Modification Methylases
  • Azacitidine