Inhibition of the eukaryotic initiation factor-2α kinase PERK decreases risk of autoimmune diabetes in mice
Charanya Muralidharan, … , Sarah A. Tersey, Raghavendra G. Mirmira
Charanya Muralidharan, … , Sarah A. Tersey, Raghavendra G. Mirmira
Published June 18, 2024
Citation Information: J Clin Invest. 2024;134(16):e176136. https://doi.org/10.1172/JCI176136.
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Research Article Endocrinology

Inhibition of the eukaryotic initiation factor-2α kinase PERK decreases risk of autoimmune diabetes in mice

Abstract

Preventing the onset of autoimmune type 1 diabetes (T1D) is feasible through pharmacological interventions that target molecular stress–responsive mechanisms. Cellular stresses, such as nutrient deficiency, viral infection, or unfolded proteins, trigger the integrated stress response (ISR), which curtails protein synthesis by phosphorylating eukaryotic translation initiation factor-2α (eIF2α). In T1D, maladaptive unfolded protein response (UPR) in insulin-producing β cells renders these cells susceptible to autoimmunity. We found that inhibition of the eIF2α kinase PKR-like ER kinase (PERK), a common component of the UPR and ISR, reversed the mRNA translation block in stressed human islets and delayed the onset of diabetes, reduced islet inflammation, and preserved β cell mass in T1D-susceptible mice. Single-cell RNA-Seq of islets from PERK-inhibited mice showed reductions in the UPR and PERK signaling pathways and alterations in antigen-processing and presentation pathways in β cells. Spatial proteomics of islets from these mice showed an increase in the immune checkpoint protein programmed death-ligand 1 (PD-L1) in β cells. Golgi membrane protein 1, whose levels increased following PERK inhibition in human islets and EndoC-βH1 human β cells, interacted with and stabilized PD-L1. Collectively, our studies show that PERK activity enhances β cell immunogenicity and that inhibition of PERK may offer a strategy for preventing or delaying the development of T1D.

Authors

Charanya Muralidharan, Fei Huang, Jacob R. Enriquez, Jiayi E. Wang, Jennifer B. Nelson, Titli Nargis, Sarah C. May, Advaita Chakraborty, Kayla T. Figatner, Svetlana Navitskaya, Cara M. Anderson, Veronica Calvo, David Surguladze, Mark J. Mulvihill, Xiaoyan Yi, Soumyadeep Sarkar, Scott A. Oakes, Bobbie-Jo M. Webb-Robertson, Emily K. Sims, Kirk A. Staschke, Decio L. Eizirik, Ernesto S. Nakayasu, Michael E. Stokes, Sarah A. Tersey, Raghavendra G. Mirmira

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Figure 5

PERK inhibition increases GOLM1 levels in β cells.

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PERK inhibition increases GOLM1 levels in β cells. (A) Representative im...
(A) Representative images of pancreata from 8-week-old female CD1, NSG, and NOD mice immunostained as indicated. Scale bars: 50 μm. Dotted lines indicate islets. (B) Quantification of the GOLM1 fluorescence intensity from data in A; each dot represents an islet. n = 3 mice (distinguished by color), with mean values for each mouse shown (ANOVA for means). (C) Representative images from 6-, 8-, 10-, and 12-week-old female NOD mice immunostained for GOLM1 (magenta), insulin (cyan), and nuclei (blue). Scale bars: 50 μm. Dotted lines indicate islets. (D) Quantification of GOLM1 fluorescence intensity from data in C; each dot represents an islet from n = 3–4 mice (distinguished by color), with mean values of each mouse shown (ANOVA for means). (E) Representative images of pancreata from 8-week-old female NOD mice that have been treated with vehicle or HC-5770 for 2 weeks, immunostained as indicated. Scale bars: 50 μm. Dotted lines indicate islets. (F) Quantification of the GOLM1 fluorescence intensity from data in E; each dot represents an islet from n = 4 mice (distinguished by color), with mean values of each mouse shown (t test for means). (G) Dot plot analysis of scRNA-Seq data in the HPAP of residual β cells for GOLM1 and CD274. The size of the dots indicates the percentage of cells that express the studied gene. The color scale shows the change of normalized and centered average gene expression within the different groups. No diabetes (ND): n = 15; single autoantibody positive (AAb1+): n = 8; double-autoantibody positive (AAb2+): n = 2; T1D: n = 9. Data are represented as mean ± SEM.