Research ArticleEndocrinology Open Access | 10.1172/JCI176136

Charanya Muralidharan,1 Fei Huang,1 Jacob R. Enriquez,1 Jiayi E. Wang,1 Jennifer B. Nelson,1 Titli Nargis,1 Sarah C. May,1 Advaita Chakraborty,1 Kayla T. Figatner,1 Svetlana Navitskaya,1 Cara M. Anderson,1 Veronica Calvo,2 David Surguladze,2 Mark J. Mulvihill,2 Xiaoyan Yi,3 Soumyadeep Sarkar,4 Scott A. Oakes,5 Bobbie-Jo M. Webb-Robertson,4 Emily K. Sims,6 Kirk A. Staschke,7 Decio L. Eizirik,3 Ernesto S. Nakayasu,4 Michael E. Stokes,2 Sarah A. Tersey,1 and Raghavendra G. Mirmira1

1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

Find articles by Stokes, M. in: JCI | PubMed | Google Scholar

1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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1Department of Medicine and the Kovler Diabetes Center, The University of Chicago, Chicago, Illinois, USA.

2HiberCell Inc., New York, New York, USA.

3ULB Center for Diabetes Research, Université Libre de Bruxelles, Brussels, Belgium.

4Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.

5Department of Pathology, The University of Chicago, Chicago, Illinois, USA.

6Department of Pediatrics, Center for Diabetes and Metabolic Diseases, Wells Center for Pediatric Research, and

7Department of Biochemistry and Molecular Biology and the Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Raghavendra G. Mirmira, 900 E. 57th Street, KCBD 8130, Chicago, Illinois 60637, USA. Phone: 773.702.2210; Email: mirmira@uchicago.edu.

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Published June 18, 2024 - More info

Published in Volume 134, Issue 16 on August 15, 2024
J Clin Invest. 2024;134(16):e176136. https://doi.org/10.1172/JCI176136.
© 2024 Muralidharan et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published June 18, 2024 - Version history
Received: October 11, 2023; Accepted: June 13, 2024 View PDF 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.

Graphical Abstract Introduction

Type 1 diabetes (T1D) is a disorder of glucose homeostasis that results from the autoimmune destruction of insulin-producing islet β cells. The importance of the immune system in initiating the early phases of T1D is emphasized in recent clinical studies showing that blockade of the T cell receptor reduces β cell stress and delays the development of T1D (1–3). These and related preclinical studies have collectively served as an impetus to shift therapeutic emphasis toward disease modification and prevention (4, 5). Molecular alterations in β cells, genetically predetermined and/or initiated by environmental insults, may contribute to early disease pathogenesis by transmitting signals that initiate and/or amplify the autoimmune assault (6, 7). Environmental insults that can trigger β cell dysfunction and T1D in individuals with genetic predispositions include, among others, viral infections, systemic inflammation, and dietary factors that all alter immune tolerance (8). As a response to these insults, various stress response mechanisms, such as the unfolded protein response (UPR), integrated stress response (ISR), autophagy, antioxidant response, and proteasomal degradation, are employed by β cells in an attempt to maintain cellular homeostasis (9).

The ISR is an evolutionarily conserved adaptive response used to mitigate cellular stress by reducing protein production burden, enhancing the expression of stress-response genes such as chaperones, and inducing the degradation of misfolded proteins (10). As part of the ISR, 4 kinases act as sensors of distinct stress signals: PKR (induced by viral infections), PKR-like ER kinase (PERK) (induced by UPR), GCN2 (induced by nutrient deprivation), and HRI (induced by Heme deprivation). When activated, each kinase phosphorylates eukaryotic translation initiation factor-2α (eIF2α) (11), which results in sequestration of initiation factor complex eIF2B. This sequestration suppresses the translation initiation of capped mRNAs while facilitating the alternative translation of “privileged” mRNAs that serve to combat stress and promote cell survival (12, 13). The translationally repressed mRNAs and their associated proteins aggregate to form nonmembranous bodies known as stress granules, where they reside until either disassembly (post stress) or removal by autophagy (persistent stress) (14). The adaptive nature of the ISR during embryogenesis is exemplified by Wolcott-Rallison syndrome, a human disorder in which homozygous loss-of-function mutations in the gene encoding PERK (EIF2AK3) result in neonatal diabetes (15); this phenotype is mirrored in Eif2ak3–/– mice (16). However, the ISR may also become maladaptive, particularly in the context of disease, and thereby exacerbate disease pathogenesis. For example, heterozygous deletion of Eif2ak3 in Akita mutant mice (which develop β cell loss and diabetes owing to a mutation in proinsulin that cripples its folding) significantly delays diabetes onset, a phenotype replicated by use of low-dose PERK inhibitors in these mice (17).

Whereas a maladaptive role for the β cell ISR during autoimmune T1D pathogenesis remains speculative, recent studies have shown the dysregulation of ISR genes in pancreatic tissue sections from donors with T1D and pre-T1D (18). The ISR kinase PERK is activated as 1 of 3 branches of the UPR. The roles of the other 2 UPR branches (ATF6 and IRE1α) have been studied in the context of T1D (19–21). Given the developmentally essential role of PERK in both the exocrine pancreas and β cell (16, 22, 23), it remains unknown whether and how PERK activity might contribute to the pathogenesis of T1D. We hypothesized that prolonged activation of PERK contributes to β cell dysfunction and maintenance of autoimmunity in T1D. In this study, we utilized a small molecule kinase inhibitor of PERK to define the molecular effects of the ISR and its role in mouse and human T1D pathogenesis. Our findings provide evidence that the ISR, via PERK, governs a molecular response that increases susceptibility of β cells to autoimmune attack and provides an alternative approach to intervening during the early stages of T1D to promote disease prevention and modification.

Results

The ISR is activated in β cells of prediabetic NOD mice and humans. ER stress in β cells has been implicated in promoting T1D pathogenesis (24). In response to ER stress, the β cell activates the UPR, in part to reduce protein load and recover ER homeostasis. To assess UPR activation in islets in the pre-T1D period, we first reanalyzed a publicly available single-cell RNA-Seq (scRNA-Seq) data set (25) of pancreatic islets isolated from female NOD mice during the prediabetic period (4, 8, and 15 weeks of age) (Figure 1A). Because the data set was enriched for immune cell populations, our analysis focused on the endocrine cell subset (mainly composed of β cells) without stratifying individual cell types. Gene set enrichment analysis (GSEA) revealed a gradual enrichment of genes of the UPR with advancing age in the endocrine cell population (Figure 1B). To specifically assess β cells, we reanalyzed a different publicly available scRNA-Seq data set (26) of islets from prediabetic female NOD mice (8, 14, and 16 weeks of age) (Figure 1C). Similarly to our observation with the endocrine cell population, GSEA showed an enrichment of the UPR over time in β cells (Figure 1D). Because the UPR includes a molecular arm (via PERK) that activates the ISR, we next probed for a hallmark feature of the ISR, phosphorylated (Ser51)-eIF2α (p-eIF2α) in pancreatic tissue of female NOD mice between 6 and 12 weeks of age. Although it did not reach statistical significance, relatively higher p-eIF2α immunostaining in β cells was observed in 6- and 8-week-old mice compared with 10- and 12-week-old mice (Supplemental Figure 1, A and B; supplemental material available online with this article; https://doi.org/10.1172/JCI176136DS1); further studies were therefore focused on NOD mice before 10 weeks of age. Because male NOD mice develop diabetes at a lower frequency than females, we compared the relative p-eIF2α immunostaining in males and females at 8 weeks of age, but observed no differences (Supplemental Figure 1, C and D). This finding suggests that activation of the ISR at this early age does not account for differences in diabetes incidence between the sexes.

Figure 1

The UPR and ISR are active in prediabetic NOD mice and proinflammatory cytokine–treated human islets. (A) UMAP embeddings of a reanalysis of scRNA-Seq of islets from female NOD mice. (B) GSEA of the endocrine cell population identified in A for UPR. (C) UMAP embeddings of a reanalysis of scRNA-Seq of islets from female NOD mice. (D) GSEA of the β cell population i