title: Engineering a glucose-responsive human insulin-secreting cell line from islets of Langerhans isolated from a patient with persistent hyperinsulinemic hypoglycemia of infancy.
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.
authors.affiliation: Department of Molecular Biology, University of Aberdeen, Institute of Medical Sciences, Aberdeen AB25 2ZD, United Kingdom.
authors.forename: W M
authors.initials: WM
authors.lastname: MacFarlane
authors.forename: J C
authors.initials: JC
authors.lastname: Chapman
authors.forename: R M
authors.initials: RM
authors.lastname: Shepherd
authors.forename: M N
authors.initials: MN
authors.lastname: Hashmi
authors.forename: N
authors.initials: N
authors.lastname: Kamimura
authors.forename: K E
authors.initials: KE
authors.lastname: Cosgrove
authors.forename: R E
authors.initials: RE
authors.lastname: O'Brien
authors.forename: P D
authors.initials: PD
authors.lastname: Barnes
authors.forename: A W
authors.initials: AW
authors.lastname: Hart
authors.forename: H M
authors.initials: HM
authors.lastname: Docherty
authors.forename: K J
authors.initials: KJ
authors.lastname: Lindley
authors.forename: A
authors.initials: A
authors.lastname: Aynsley-Green
authors.forename: R F
authors.initials: RF
authors.lastname: James
authors.forename: K
authors.initials: K
authors.lastname: Docherty
authors.forename: M J
authors.initials: MJ
authors.lastname: Dunne
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005818:Genetic Engineering; D005947:Glucose / Q000494:pharmacology*; D018398:Homeodomain Proteins; D006801:Humans; D006946:Hyperinsulinism / Q000235:genetics* / Q000473:pathology; D007003:Hypoglycemia / Q000235:genetics* / Q000473:pathology; D007223:Infant; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D015221:Potassium Channels / Q000235:genetics; D024661:Potassium Channels, Inwardly Rectifying; D011189:Potassium Chloride / Q000494:pharmacology; D011955:Receptors, Drug / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D064233:Sulfonylurea Receptors; D014044:Tolbutamide / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: C577507:ABCC8 protein, human; D018528:ATP-Binding Cassette Transporters; C577508:Abcc8 protein, mouse; D018398:Homeodomain Proteins; D007328:Insulin; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D011955:Receptors, Drug; D011993:Recombinant Fusion Proteins; D064233:Sulfonylurea Receptors; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D014044:Tolbutamide; D005947:Glucose; D002118:Calcium