Functional Proteasome Complex Is Required for Turnover of Islet Amyloid Polypeptide in Pancreatic β-cells

J Biol Chem. 2018 Sep 14;293(37):14210-14223. doi: 10.1074/jbc.RA118.002414. Epub 2018 Jul 16.

Abstract

Human islet amyloid polypeptide (hIAPP) is the principal constituent of amyloid deposits and toxic oligomers in the pancreatic islets. Together with hyperglycemia, hIAPP-derived oligomers and aggregates are important culprits in type 2 diabetes mellitus (T2DM). Here, we explored the role of the cell's main proteolytic complex, the proteasome, in hIAPP turnover in normal and stressed β-cells evoked by chronic hyperglycemia. Moderate inhibition (10-35%) of proteasome activity/function in cultured human islets by the proteasome inhibitor lactacystin enhanced intracellular accumulation of hIAPP. Unexpectedly, prolonged (>1 h) and marked (>50%) impairment of proteasome activity/function had a strong inhibitory effect on hIAPP transcription and secretion from normal and stressed β-cells. This negative compensatory feedback mechanism for controlling IAPP turnover was also observed in the lactacystin-treated rat insulinoma β-cell line (INS 832/13), demonstrating the presence of an evolutionarily conserved mechanism for IAPP production. In line with these in situ studies, our current ex vivo data showed that proteasome activity and hIAPP expression are also down-regulated in islets isolated from T2DM subjects. Gene expression and promoter activity studies demonstrated that the functional proteasome complex is required for efficient activation of the hIAPP promoter and for full expression of IAPP's essential transcription factor, FOXA2. ChIP studies revealed that promoter occupancy of FoxA2 at the rat IAPP promoter region is an important and limiting factor for amylin expression in proteasome-impaired murine cells. This study suggests a novel regulatory pathway in β-cells involving proteasome, FOXA2, and IAPP, which can be possibly targeted to regulate hIAPP levels and islet amyloidosis in T2DM.

Keywords: FoxA2; Islet amyloid polypeptide; amyloid; hyperglycemia; insulin; proteasome; secretion; transcription; type 2 diabetes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetylcysteine / analogs & derivatives
  • Acetylcysteine / pharmacology
  • Animals
  • Cell Line, Tumor
  • Diabetes Mellitus, Type 2 / metabolism*
  • Diabetes Mellitus, Type 2 / pathology
  • Down-Regulation
  • Hepatocyte Nuclear Factor 3-beta / metabolism
  • Humans
  • Insulin-Secreting Cells / enzymology
  • Insulin-Secreting Cells / metabolism*
  • Insulinoma / enzymology
  • Insulinoma / metabolism
  • Islet Amyloid Polypeptide / genetics
  • Islet Amyloid Polypeptide / metabolism*
  • Leupeptins / pharmacology
  • Mice
  • Oligopeptides / pharmacology
  • Promoter Regions, Genetic
  • Proteasome Endopeptidase Complex / drug effects
  • Proteasome Endopeptidase Complex / metabolism*
  • Proteasome Inhibitors / pharmacology
  • Rats

Substances

  • FOXA2 protein, human
  • Islet Amyloid Polypeptide
  • Leupeptins
  • Oligopeptides
  • Proteasome Inhibitors
  • lactacystin
  • Hepatocyte Nuclear Factor 3-beta
  • Proteasome Endopeptidase Complex
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde
  • Acetylcysteine
  • epoxomicin