Spatiotemporal Correlation Spectroscopy Reveals a Protective Effect of Peptide-Based GLP-1 Receptor Agonism against Lipotoxicity on Insulin Granule Dynamics in Primary Human β-Cells.

Gianmarco Ferri,Francesco Cardarelli,Luca Pesce,Mara Suleiman,Marta Tesi,Carmela De Luca,Lorella Marselli,Margherita Occhipinti,Piero Marchetti,Francesca Grano,Marco Bugliani

doi:10.3390/pharmaceutics13091403

Abstract

Glucagon-like peptide-1 receptor (GLP-1R) agonists are being used for the treatment of type 2 diabetes (T2D) and may have beneficial effects on the pancreatic β-cells. Here, we evaluated the effects of GLP-1R agonism on insulin secretory granule (ISG) dynamics in primary β-cells isolated from human islets exposed to palmitate-induced lipotoxic stress. Islets cells were exposed for 48 h to 0.5 mM palmitate (hereafter, ‘Palm’) with or without the addition of a GLP-1 agonist, namely 10 nM exendin-4 (hereafter, ‘Ex-4’). Dissociated cells were first transfected with syncollin-EGFP in order to fluorescently mark the ISGs. Then, by applying a recently established spatiotemporal correlation spectroscopy technique, the average structural (i.e., size) and dynamic (i.e., the local diffusivity and mode of motion) properties of ISGs are extracted from a calculated imaging-derived Mean Square Displacement (iMSD) trace. Besides defining the structural/dynamic fingerprint of ISGs in human cells for the first time, iMSD analysis allowed to probe fingerprint variations under selected conditions: namely, it was shown that Palm affects ISGs dynamics in response to acute glucose stimulation by abolishing the ISGs mobilization typically imparted by glucose and, concomitantly, by reducing the extent of ISGs active/directed intracellular movement. By contrast, co-treatment with Ex-4 normalizes ISG dynamics, i.e., re-establish ISG mobilization and ability to perform active transport in response to glucose stimulation. These observations were correlated with standard glucose-stimulated insulin secretion (GSIS), which resulted in being reduced in cells exposed to Palm but preserved in cells concomitantly exposed to 10 nM Ex-4. Our data support the idea that GLP-1R agonism may exert its beneficial effect on human β-cells under metabolic stress by maintaining ISGs’ proper intracellular dynamics.

Highlights

Here we propose the application of the imaging-derived Mean Square Displacement (iMSD) approach to evaluate the effect of palmitate-induced lipotoxicity (‘Palm’ treatment) and potential protection by the GLP-1 agonist Exendin-4 (‘Ex-4’ treatment) on insulin secretory granule (ISG) in primary living β-cells dissociated from human pancreatic islet
Co-treatment with Ex-4 normalizes ISG dynamics, i.e., re-establish ISG mobilization and ability to perform active transport in response to glucose. These effects are correlated with standard glucose-stimulated insulin secretion (GSIS), which resulted in being significantly reduced in cells exposed to Palm but preserved in cells concomitantly exposed to 10 nM Ex-4
Normalized distributions extracted by fitting procedures of size, Dm, and α are plotted in Figure 1b and compared to data obtained from -labeled ISGs in INS-1E cells

Summary

Introduction

Pancreatic β-cell dysfunction, determined by the interplay of genetic and acquired factors, has a major role in the development and progression of type 2 diabetes (T2D) [1,2]. In this regard, increased concentrations of certain fatty acids (palmitate in particular) have been shown to induce a lipotoxic effect and impair β-cell function, survival and even proliferation [3,4,5]. Among the several pharmacological treatments for the therapy of diabetes, GLP-1 receptor (GLP-1R) agonists are being used, with a favorable benefit/risk ratio [6,7] These compounds, have several beneficial actions, including possible

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