Engineering a glucose-responsive human insulin-secreting cell line from islets of langerhans isolated from a patient with persistent hyperinsulinemichypoglycemia of infancy

Persistent hyperinsulinemic hypoglycemia of infancy (PHHI) is a neonatal di sease characterized by dysregulation of insulin secretion accompanied by pr ofound hypoglycemia, We have discovered that islet cells, isolated from the pancreas of a PHHI patient, proliferate in culture while maintaining a bet a cell-like phenotype, The PHHI-derived cell line (NES2Y) exhibits insulin secretory characteristics typical of islet cells derived from these patient s, i,e, they have no K-ATP channel activity and as a consequence secrete in sulin at constitutively high levels in the absence of glucose, In addition, they exhibit impaired expression of the homeodomain transcription factor P DX1, which is a key component of the signaling pathway linking nutrient met abolism to the regulation of insulin gene expression. To repair these defec ts NES2Y cells were triple-transfected with cDNAs encoding the two componen ts of the K-ATP channel (SUR1 and Kir6.2) and PDX1, One selected clonal cel l line (NISK9) had normal K-ATP channel activity, and as a result of change s in intracellular Ca2+ homeostasis ([Ca2+](i)) secreted insulin within the physiological range of glucose concentrations. This approach to engineerin g PHHI-derived islet cells may be of use in gene therapy for PHHI and in ce ll engineering techniques for administering insulin for the treatment of di abetes mellitus.