Engineering a glucose-responsive human insulin-secreting cell line from islets of Langerhans isolated from a patient with persistent hyperinsulinemic hypoglycemia of infancy

W M MacFarlane, J C Chapman, R M Shepherd, M N Hashmi, N Kamimura, K E Cosgrove, R E O'Brien, P D Barnes, Alan Hart, H M Docherty, K J Lindley, A Aynsley-Green, R F James, K Docherty, M J Dunne

Research output: Contribution to journalArticlepeer-review

Abstract

Persistent hyperinsulinemic hypoglycemia of infancy (PHHI) is a neonatal disease characterized by dysregulation of insulin secretion accompanied by profound hypoglycemia. We have discovered that islet cells, isolated from the pancreas of a PHHI patient, proliferate in culture while maintaining a beta cell-like phenotype. The PHHI-derived cell line (NES2Y) exhibits insulin secretory characteristics typical of islet cells derived from these patients, i.e. they have no K(ATP) channel activity and as a consequence secrete insulin at constitutively high levels in the absence of glucose. In addition, they exhibit impaired expression of the homeodomain transcription factor PDX1, which is a key component of the signaling pathway linking nutrient metabolism to the regulation of insulin gene expression. To repair these defects NES2Y cells were triple-transfected with cDNAs encoding the two components of the K(ATP) channel (SUR1 and Kir6.2) and PDX1. One selected clonal cell line (NISK9) had normal K(ATP) channel activity, and as a result of changes in intracellular Ca(2+) homeostasis ([Ca(2+)](i)) secreted insulin within the physiological range of glucose concentrations. This approach to engineering PHHI-derived islet cells may be of use in gene therapy for PHHI and in cell engineering techniques for administering insulin for the treatment of diabetes mellitus.
Original languageEnglish
Pages (from-to)34059-66
Number of pages8
JournalJournal of Biological Chemistry
Volume274
Issue number48
DOIs
Publication statusPublished - 26 Nov 1999

Keywords

  • Animals
  • Calcium
  • Humans
  • Glucose
  • Islets of Langerhans
  • Hypoglycemia
  • Insulin
  • Potassium Channels
  • Potassium Chloride
  • Infant
  • Hyperinsulinism
  • Tumor Cells, Cultured
  • Receptors, Drug
  • Potassium Channels, Inwardly Rectifying
  • Adenosine Triphosphate
  • Tolbutamide
  • Trans-Activators
  • Homeodomain Proteins
  • Genetic Engineering
  • Dose-Response Relationship, Drug
  • Mice
  • Recombinant Fusion Proteins
  • Electrophysiology
  • Adenosine Diphosphate
  • Transfection
  • ATP-Binding Cassette Transporters
  • Membrane Potentials
  • Cell Line

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