A Targeted RNAi Screen Identifies Endocytic Trafficking Factors That Control GLP-1 Receptor Signaling in Pancreatic β-Cells

Diabetes. 2018 Mar;67(3):385-399. doi: 10.2337/db17-0639. Epub 2017 Dec 28.

Abstract

The glucagon-like peptide 1 (GLP-1) receptor (GLP-1R) is a key target for type 2 diabetes (T2D) treatment. Because endocytic trafficking of agonist-bound receptors is one of the most important routes for regulation of receptor signaling, a better understanding of this process may facilitate the development of new T2D therapeutic strategies. Here, we screened 29 proteins with known functions in G protein-coupled receptor trafficking for their role in GLP-1R potentiation of insulin secretion in pancreatic β-cells. We identify five (clathrin, dynamin1, AP2, sorting nexins [SNX] SNX27, and SNX1) that increase and four (huntingtin-interacting protein 1 [HIP1], HIP14, GASP-1, and Nedd4) that decrease insulin secretion from murine insulinoma MIN6B1 cells in response to the GLP-1 analog exendin-4. The roles of HIP1 and the endosomal SNX1 and SNX27 were further characterized in mouse and human β-cell lines and human islets. While HIP1 was required for the coupling of cell surface GLP-1R activation with clathrin-dependent endocytosis, the SNXs were found to control the balance between GLP-1R plasma membrane recycling and lysosomal degradation and, in doing so, determine the overall β-cell incretin responses. We thus identify key modulators of GLP-1R trafficking and signaling that might provide novel targets to enhance insulin secretion in T2D.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium Signaling / drug effects
  • Cell Line
  • Cyclic AMP / metabolism
  • DNA-Binding Proteins / antagonists & inhibitors
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Endocytosis* / drug effects
  • Exenatide
  • Glucagon-Like Peptide-1 Receptor / agonists*
  • Glucagon-Like Peptide-1 Receptor / genetics
  • Glucagon-Like Peptide-1 Receptor / metabolism
  • Humans
  • Incretins / pharmacology
  • Insulin / metabolism
  • Insulin Secretion
  • Insulin-Secreting Cells / drug effects
  • Insulin-Secreting Cells / metabolism*
  • Insulin-Secreting Cells / ultrastructure
  • Lysosomes / drug effects
  • Lysosomes / enzymology
  • Lysosomes / metabolism
  • Lysosomes / ultrastructure
  • Mice
  • Microscopy, Electron, Transmission
  • Peptides / pharmacology
  • RNA Interference
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Second Messenger Systems* / drug effects
  • Sorting Nexins / antagonists & inhibitors
  • Sorting Nexins / genetics
  • Sorting Nexins / metabolism*
  • Tissue Culture Techniques
  • Venoms / pharmacology

Substances

  • DNA-Binding Proteins
  • GLP1R protein, human
  • Glp1r protein, mouse
  • Glucagon-Like Peptide-1 Receptor
  • HIP1 protein, human
  • Hip1 protein, mouse
  • Incretins
  • Insulin
  • Peptides
  • Recombinant Fusion Proteins
  • SNX1 protein, human
  • SNX1 protein, mouse
  • SNX27 protein, human
  • Snx27 protein, mouse
  • Sorting Nexins
  • Venoms
  • Exenatide
  • Cyclic AMP