Impaired expression of transcription factor IUF1 in a pancreatic β‐cell line derived from a patient with persistent hyperinsulinaemic hypoglycaemia of infancy (nesidioblastosis)

Abstract

Persistent hyperinsulinaemic hypoglycaemia of infancy (PHHI), or nesidioblastosis, is a rare disorder which may be familial or sporadic, and which is characterized by unregulated secretion of insulin and profound hypoglycaemia in the neonate. The defect has been linked in some patients to mutations in the sulphonyl urea receptor gene (SUR). The present study investigated potential defects in the regulation of the insulin gene by glucose in a P-cell line (NES 2Y) derived from a patient with PHHI. The results show that the insulin promoter is unresponsive to glucose in PHHI, and that this defect can be attributed to impaired expression of the transcription factor IUF1. Because IUF1 is involved not only in linking glucose metabolism to the control of the insulin, but is also a major regulator of P-cell differentiation during embryogenesis, we propose that impaired expression of IUF1 contributes to P-cell dysfunction in PHHI by leading to abnormal P-cell differentiation.© 1997 Federation of European Biochemical Societies.

Involvement of Per–Arnt–Sim (PAS) kinase in the stimulation of preproinsulin and pancreatic duodenum homeobox 1 gene expression by glucose

“…MIN6 ␤ cells (27) (passages [19][20][21][22][23][24][25][26][27][28][29][30] were grown in DMEM containing 15% FCS, 25 mM glucose, 5.4 mM KCl, 2 mM glutamine, 100 mM 2-mercaptoethanol, 100 units͞ml Ϫ1 penicillin, and 100 g͞ml Ϫ1 streptomycin in a humidified atmosphere at 37°C with 5% CO 2 , and seeded onto poly(L)-lysinecoated coverslips for microinjection. Cells were transfected with Lipofectamine 2000 or TransIT-TKO (for siRNA).…”

Characterisation of the NES2Y cell line and its use in the production of human glucose-responsive insulin producing (hGRIP) cell lines by cell-cell fusion

“…25 The passage of the cells occurred over 15 stages and this cell line had the crucial characteristics of being able to grow in culture but contained defective potassium channel expression indicative of the original disease; thus was not glucose responsive. Therefore, we considered this to be an appropriate candidate for producing functional β-cell hybrids.…”

“…31,32 In 1997 Macfarlane et al described the derivation of a human insulin-producing cell line, which showed defective potassium channel expression and was not glucose responsive but, crucially, had characteristics of being able to grow in culture. 25 In this work, flow cytometric analysis was carried out to examine the intracellular hormone and other endocrine cell-related antigenic expression of NES2Y cells from primary culture. The results showed that the NES2Y cells exhibited an endocrine phenotype with continuous culture over 2, 4, 6 and 8 months.…”

“…NES2Y cells were derived from islets of Langerhans isolated from the pancreas of a patient with PHHI as described previously. 25 Cells were maintained under standard tissue culture conditions using RPMI-1640 medium (Sigma, Poole, UK) supplemented with 10% (vol/vol) fetal calf serum, 100 U/ml penicillin and 100 μg/ml streptomycin in a humidified atmosphere of 5% CO 2 in air at 37°C.…”

“…In order to achieve this goal we have utilized the NES2Y cell line which was originally described by MacFarlane et al 25,26 where NES2Y cells grow continuously in culture and can, therefore, be considered to be immortalized. In addition, the NES2Y cell line exhibited insulin secretion in vitro but with defective K ATP Diabetes mellitus is a debilitating disease and alternative methods of treatment are a priority if the short-term and long term sequelae are to be avoided.…”