1. Engineering a Glucose-responsive Human Insulin-secreting Cell Line from Islets of Langerhans Isolated from a Patient with Persistent Hyperinsulinemic Hypoglycemia of Infancy
- Author
-
Mark J. Dunne, Hilary M. Docherty, Rachel E. O'Brien, Alan W. Hart, Wendy M. Macfarlane, Joanna C. Chapman, RM Shepherd, Philippa D. Barnes, Noritaka Kamimura, Keith J. Lindley, Kevin Docherty, Karen E. Cosgrove, Albert Aynsley-Green, MN Hashmi, and R. F. L. James
- Subjects
Potassium Channels ,Receptors, Drug ,medicine.medical_treatment ,Sulfonylurea Receptors ,medicine.disease_cause ,Biochemistry ,Membrane Potentials ,Potassium Chloride ,Mice ,Adenosine Triphosphate ,Insulin Secretion ,Tumor Cells, Cultured ,Insulin ,geography.geographical_feature_category ,Islet ,Insulin oscillation ,Adenosine Diphosphate ,Electrophysiology ,PDX1 ,Genetic Engineering ,medicine.drug ,endocrine system ,medicine.medical_specialty ,Recombinant Fusion Proteins ,Tolbutamide ,Hypoglycemia ,Biology ,Transfection ,Cell Line ,Islets of Langerhans ,Hyperinsulinism ,Internal medicine ,medicine ,Animals ,Humans ,Potassium Channels, Inwardly Rectifying ,Hyperinsulinemic hypoglycemia ,Molecular Biology ,Homeodomain Proteins ,geography ,Dose-Response Relationship, Drug ,Infant ,Cell Biology ,medicine.disease ,Glucose ,Endocrinology ,Trans-Activators ,ATP-Binding Cassette Transporters ,Calcium - 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.
- Published
- 1999
- Full Text
- View/download PDF