TCF7L2 Regulates Late Events in Insulin Secretion From Pancreatic Islet β-Cells

TCF7L2 Regulates Late Events in Insulin Secretion From Pancreatic Islet β-Cells Gabriela da Silva Xavier 1 , Merewyn K. Loder 1 , Angela McDonald 1 , Andrei I. Tarasov 1 , Raffaella Carzaniga 2 , Katrin Kronenberger 2 , Sebastian Barg 3 and Guy A. Rutter 1 1 Section of Cell Biology, Division of Medicine, Faculty of Medicine, Imperial College, London, U.K.; 2 Electron Microscopy Centre, Imperial College London, South Kensington, U.K.; 3 Medical Cell Biology, Uppsala University, Uppsala, Sweden. Corresponding author: Guy A. Rutter, g.rutter{at}imperial.ac.uk . G.d.S.X., M.K.L., and A.M. contributed equally to this work. Abstract OBJECTIVE Polymorphisms in the human TCF7L2 gene are associated with reduced insulin secretion and an increased risk of type 2 diabetes. However, the mechanisms by which TCF7L2 affect insulin secretion are still unclear. We define the effects of TCF7L2 expression level on mature β-cell function and suggest a potential mechanism for its actions. RESEARCH DESIGN AND METHODS TCF7L2 expression in rodent islets and β-cell lines was altered using RNAi or adenoviral transduction. β-Cell gene profiles were measured by quantitative real-time PCR and the effects on intracellular signaling and exocytosis by live cell imaging, electron microscopy, and patch clamp electrophysiology. RESULTS Reducing TCF7L2 expression levels by RNAi decreased glucose- but not KCl-induced insulin secretion. The glucose-induced increments in both ATP/ADP ratio and cytosolic free Ca 2+ concentration ([Ca 2+ ] i ) were increased compared with controls. Overexpression of TCF7L2 exerted minor inhibitory effects on glucose-regulated changes in [Ca 2+ ] i and insulin release. Gene expression profiling in TCF7L2-silenced cells revealed increased levels of mRNA encoding syntaxin 1A but decreased Munc18–1 and ZnT8 mRNA. Whereas the number of morphologically docked vesicles was unchanged by TCF7L2 suppression, secretory granule movement increased and capacitance changes decreased, indicative of defective vesicle fusion. RESULTS CONCLUSION—TCF7L2 is involved in maintaining expression of β-cell genes regulating secretory granule fusion. Defective insulin exocytosis may thus underlie increased diabetes incidence in carriers of the at-risk TCF7L2 alleles. Footnotes The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate t