Increased hexosamine biosynthetic pathway flux alters cell-cell adhesion in INS-1E cells and murine islets

Endocrine. 2023 Sep;81(3):492-502. doi: 10.1007/s12020-023-03412-9. Epub 2023 Jun 12.

Abstract

Purpose: In type 2 Diabetes, β-cell failure is caused by loss of cell mass, mostly by apoptosis, but also by simple dysfunction (dedifferentiation, decline of glucose-stimulated insulin secretion). Apoptosis and dysfunction are caused, at least in part, by glucotoxicity, in which increased flux of glucose in the hexosamine biosynthetic pathway plays a role. In this study, we sought to clarify whether increased hexosamine biosynthetic pathway flux affects another important aspect of β-cell physiology, that is β-cell-β-cell homotypic interactions.

Methods: We used INS-1E cells and murine islets. The expression and cellular distribution of E-cadherin and β-catenin was evaluated by immunofluorescence, immunohistochemistry and western blot. Cell-cell adhesion was examined by the hanging-drop aggregation assay, islet architecture by isolation and microscopic observation.

Results: E-cadherin expression was not changed by increased hexosamine biosynthetic pathway flux, however, there was a decrease of cell surface, and an increase in intracellular E-cadherin. Moreover, intracellular E-cadherin delocalized, at least in part, from the Golgi complex to the endoplasmic reticulum. Beta-catenin was found to parallel the E-cadherin redistribution, showing a dislocation from the plasmamembrane to the cytosol. These changes had as a phenotypic consequence a decreased ability of INS-1E to aggregate. Finally, in ex vivo experiments, glucosamine was able to alter islet structure and to decrease surface abundandance of E-cadherin and β-catenin.

Conclusion: Increased hexosamine biosynthetic pathway flux alters E-cadherin cellular localization both in INS-1E cells and murine islets and affects cell-cell adhesion and islet morphology. These changes are likely caused by alterations of E-cadherin function, highlighting a new potential target to counteract the consequences of glucotoxicity on β-cells.

Keywords: E-cadherin; Murine islets; Pancreatic beta-cell.

MeSH terms

  • Animals
  • Biosynthetic Pathways
  • Cadherins / metabolism
  • Cell Adhesion
  • Diabetes Mellitus, Type 2* / metabolism
  • Glucose / metabolism
  • Hexosamines / metabolism
  • Insulin / metabolism
  • Insulin-Secreting Cells* / metabolism
  • Islets of Langerhans* / metabolism
  • Mice
  • beta Catenin / metabolism

Substances

  • Insulin
  • beta Catenin
  • Hexosamines
  • Glucose
  • Cadherins