Seven mutations in the human insulin gene linked to permanent neonatal/infancy-onset diabetes mellitus.

Permanent neonatal diabetes mellitus (PNDM) is a rare disorder usually presenting within 6 months of birth. Although several genes have been linked to this disorder, in almost half the cases documented in Italy, the genetic cause remains unknown. Because the Akita mouse bearing a mutation in the Ins2 gene exhibits PNDM associated with pancreatic beta cell apoptosis, we sequenced the human insulin gene in PNDM subjects with unidentified mutations. We discovered 7 heterozygous mutations in 10 unrelated probands. In 8 of these patients, insulin secretion was detectable at diabetes onset, but rapidly declined over time. When these mutant proinsulins were expressed in HEK293 cells, we observed defects in insulin protein folding and secretion. In these experiments, expression of the mutant proinsulins was also associated with increased Grp78 protein expression and XBP1 mRNA splicing, 2 markers of endoplasmic reticulum stress, and with increased apoptosis. Similarly transfected INS-1E insulinoma cells had diminished viability compared with those expressing WT proinsulin. In conclusion, we find that mutations in the insulin gene that promote proinsulin misfolding may cause PNDM.

Surfactant kinetics in preterm infants on mechanical ventilation who did and did not develop bronchopulmonary dysplasia.

OBJECTIVE: To characterize surfactant kinetics in vivo in two groups of premature infants on different levels of mechanical ventilation and at different risk of developing bronchopulmonary dysplasia. DESIGN: Controlled observational study in two independent groups of infants. SETTING: Neonatal intensive care unit. PATIENTS: Thirteen preterm infants (26 +/- 0.5 wks, birth weight 801 +/- 64 g) on high ventilatory setting and who finally all developed bronchopulmonary dysplasia (MechVentBPD), and eight (26 +/- 0.5 wks, birth weight 887 +/- 103 g) who had minimal or no lung disease and of whom none developed bronchopulmonary dysplasia (MechVentNoBPD). MEASUREMENTS AND MAIN RESULTS: Endotracheal 13C-labeled dipalmitoyl-phosphatidylcholine was administered and subsequent measurements of the 13C enrichment of surfactant-disaturated phosphatidylcholine (DSPC) from serial tracheal aspirates were made by gas chromatography-mass spectrometry. We calculated disaturated phosphatidylcholine pharmacokinetic variables in terms of half-life and apparent pool size from the enrichment decay curves over time. DSPC concentration from tracheal aspirates was expressed as milligrams/milliliter epithelial lining fluid (ELF-DSPC). Data are presented as mean +/- se. In MechVentBPD infants vs. MechVentNoBPD, ELF-DSPC was much reduced, 2.9 +/- 0.6 vs. 9.4 +/- 3.0 mg/mL ELF (p =.03), half-life was shorter, 19.4 +/- 2.8 vs. 42.5 +/- 6.3 hrs (p =.002), and apparent pool size larger, 136 +/- 21 vs. 65.8 +/- 16.0 mg/kg (p =.057). In MechVentBPD, apparent DSPC pool size positively correlated with mean airway pressure x Fio(2) and inversely correlated with ELF-DSPC. ELF-DSPC was inversely correlated with mean airway pressure x Fio(2). No significant correlations were found in the MechVentNoBPD group. CONCLUSIONS: MechVentBPD infants showed profound alteration of surfactant kinetics compared with preterm infants with minimal lung disease, and these alterations were correlated with severity of ventilatory support.