Author: "Helen E. Thomas" / Language: english - Searchworks@Jio Institute Digital Library Search Results

46 results on '"Helen E. Thomas"'

1. TIGIT acts as an immune checkpoint upon inhibition of PD1 signaling in autoimmune diabetes

Author: Prerak Trivedi, Gaurang Jhala, David J. De George, Chris Chiu, Claudia Selck, Tingting Ge, Tara Catterall, Lorraine Elkerbout, Louis Boon, Nicole Joller, Thomas W. Kay, Helen E. Thomas, and Balasubramanian Krishnamurthy
Subjects: type 1 diabetes, TIGIT, CD8+ T cell, immune checkpoint, NOD mouse, Immunologic diseases. Allergy, RC581-607
Abstract: IntroductionChronic activation of self-reactive T cells with beta cell antigens results in the upregulation of immune checkpoint molecules that keep self-reactive T cells under control and delay beta cell destruction in autoimmune diabetes. Inhibiting PD1/PD-L1 signaling results in autoimmune diabetes in mice and humans with pre-existing autoimmunity against beta cells. However, it is not known if other immune checkpoint molecules, such as TIGIT, can also negatively regulate self-reactive T cells. TIGIT negatively regulates the CD226 costimulatory pathway, T-cell receptor (TCR) signaling, and hence T-cell function.MethodsThe phenotype and function of TIGIT expressing islet infiltrating T cells was studied in non-obese diabetic (NOD) mice using flow cytometry and single cell RNA sequencing. To determine if TIGIT restrains self-reactive T cells, we used a TIGIT blocking antibody alone or in combination with anti-PDL1 antibody.ResultsWe show that TIGIT is highly expressed on activated islet infiltrating T cells in NOD mice. We identified a subset of stem-like memory CD8+ T cells expressing multiple immune checkpoints including TIGIT, PD1 and the transcription factor EOMES, which is linked to dysfunctional CD8+ T cells. A known ligand for TIGIT, CD155 was expressed on beta cells and islet infiltrating dendritic cells. However, despite TIGIT and its ligand being expressed, islet infiltrating PD1+TIGIT+CD8+ T cells were functional. Inhibiting TIGIT in NOD mice did not result in exacerbated autoimmune diabetes while inhibiting PD1-PDL1 resulted in rapid autoimmune diabetes, indicating that TIGIT does not restrain islet infiltrating T cells in autoimmune diabetes to the same degree as PD1. Partial inhibition of PD1-PDL1 in combination with TIGIT inhibition resulted in rapid diabetes in NOD mice. DiscussionThese results suggest that TIGIT and PD1 act in synergy as immune checkpoints when PD1 signaling is partially impaired. Beta cell specific stem-like memory T cells retain their functionality despite expressing multiple immune checkpoints and TIGIT is below PD1 in the hierarchy of immune checkpoints in autoimmune diabetes.
Published: 2024
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2. Desmoglein-2 is important for islet function and β-cell survival

Author: Kay K. Myo Min, Darling Rojas-Canales, Daniella Penko, Mark DeNichilo, Michaelia P. Cockshell, Charlie B. Ffrench, Emma J. Thompson, Olof Asplund, Christopher J. Drogemuller, Rashmi B. Prasad, Leif Groop, Shane T. Grey, Helen E. Thomas, Thomas Loudovaris, Thomas W. Kay, My G. Mahoney, Claire F. Jessup, P. Toby Coates, and Claudine S. Bonder
Subjects: Cytology, QH573-671
Abstract: Abstract Type 1 diabetes is a complex disease characterized by the lack of endogenous insulin secreted from the pancreatic β-cells. Although β-cell targeted autoimmune processes and β-cell dysfunction are known to occur in type 1 diabetes, a complete understanding of the cell-to-cell interactions that support pancreatic function is still lacking. To characterize the pancreatic endocrine compartment, we studied pancreata from healthy adult donors and investigated a single cell surface adhesion molecule, desmoglein-2 (DSG2). Genetically-modified mice lacking Dsg2 were examined for islet cell mass, insulin production, responses to glucose, susceptibility to a streptozotocin-induced mouse model of hyperglycaemia, and ability to cure diabetes in a syngeneic transplantation model. Herein, we have identified DSG2 as a previously unrecognized adhesion molecule that supports β-cells. Furthermore, we reveal that DSG2 is within the top 10 percent of all genes expressed by human pancreatic islets and is expressed by the insulin-producing β-cells but not the somatostatin-producing δ-cells. In a Dsg2 loss-of-function mice (Dsg2 lo/lo), we observed a significant reduction in the number of pancreatic islets and islet size, and consequently, there was less total insulin content per islet cluster. Dsg2 lo/lo mice also exhibited a reduction in blood vessel barrier integrity, an increased incidence of streptozotocin-induced diabetes, and islets isolated from Dsg2 lo/lo mice were more susceptible to cytokine-induced β-cell apoptosis. Following transplantation into diabetic mice, islets isolated from Dsg2 lo/lo mice were less effective than their wildtype counterparts at curing diabetes. In vitro assays using the Beta-TC-6 murine β-cell line suggest that DSG2 supports the actin cytoskeleton as well as the release of cytokines and chemokines. Taken together, our study suggests that DSG2 is an under-appreciated regulator of β-cell function in pancreatic islets and that a better understanding of this adhesion molecule may provide new opportunities to combat type 1 diabetes.
Published: 2022
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3. Monitoring immunomodulation strategies in type 1 diabetes

Author: Balasubramanian Krishnamurthy, Matthew Lacorcia, Thomas W. H. Kay, Helen E. Thomas, and Stuart I. Mannering
Subjects: type 1 diabetes, biomarkers, T cell exhaustion, antigen specific therapy, disease modifying treatment, Immunologic diseases. Allergy, RC581-607
Abstract: Type 1 diabetes (T1D) is a T-cell mediated autoimmune disease. Short-term treatment with agents targeting T cells, B cells and inflammatory cytokines to modify the disease course resulted in a short-term pause in disease activity. Lessons learnt from these trials will be discussed in this review. It is expected that effective disease-modifying agents will become available for use in earlier stages of T1D. Progress has been made to analyze antigen-specific T cells with standardization of T cell assay and discovery of antigen epitopes but there are many challenges. High-dimensional profiling of gene, protein and TCR expression at single cell level with innovative computational tools should lead to novel biomarker discovery. With this, assays to detect, quantify and characterize the phenotype and function of antigen-specific T cells will continuously evolve. An improved understanding of T cell responses will help researchers and clinicians to better predict disease onset, and progression, and the therapeutic efficacy of interventions to prevent or arrest T1D.
Published: 2023
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4. Inflammation versus regulation: how interferon-gamma contributes to type 1 diabetes pathogenesis

Author: David J. De George, Tingting Ge, Balasubramaniam Krishnamurthy, Thomas W. H. Kay, and Helen E. Thomas
Subjects: type 1 daibetes mellitus, interferon gamma (IFNγ), CD8 T cells, inflammation, autoimmunity, T cell proliferation, Biology (General), QH301-705.5
Abstract: Type 1 diabetes is an autoimmune disease with onset from early childhood. The insulin-producing pancreatic beta cells are destroyed by CD8+ cytotoxic T cells. The disease is challenging to study mechanistically in humans because it is not possible to biopsy the pancreatic islets and the disease is most active prior to the time of clinical diagnosis. The NOD mouse model, with many similarities to, but also some significant differences from human diabetes, provides an opportunity, in a single in-bred genotype, to explore pathogenic mechanisms in molecular detail. The pleiotropic cytokine IFN-γ is believed to contribute to pathogenesis of type 1 diabetes. Evidence of IFN-γ signaling in the islets, including activation of the JAK-STAT pathway and upregulation of MHC class I, are hallmarks of the disease. IFN-γ has a proinflammatory role that is important for homing of autoreactive T cells into islets and direct recognition of beta cells by CD8+ T cells. We recently showed that IFN-γ also controls proliferation of autoreactive T cells. Therefore, inhibition of IFN-γ does not prevent type 1 diabetes and is unlikely to be a good therapeutic target. In this manuscript we review the contrasting roles of IFN-γ in driving inflammation and regulating the number of antigen specific CD8+ T cells in type 1 diabetes. We also discuss the potential to use JAK inhibitors as therapy for type 1 diabetes, to inhibit both cytokine-mediated inflammation and proliferation of T cells.
Published: 2023
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5. Integrin αvβ5 heterodimer is a specific marker of human pancreatic beta cells

Author: Jacqueline V. Schiesser, Thomas Loudovaris, Helen E. Thomas, Andrew G. Elefanty, and Edouard G. Stanley
Subjects: Medicine, Science
Abstract: Abstract The identification of cell surface markers specific to pancreatic beta cells is important for both the study of islet biology and for investigating the pathophysiology of diseases in which this cell type is lost or damaged. Following analysis of publicly available RNAseq data, we identified specific integrin subunits, integrin αv and integrin β5, that were expressed in beta cells. This finding was further elaborated using immunofluorescence analysis of histological sections derived from donor human pancreas. Despite the broad expression of specific integrin subunits, we found that expression of integrin αvβ5 heterodimers was restricted to beta cells and that this complex persisted in islet remnants of some type 1 diabetic individuals from which insulin expression had been lost. This study identifies αvβ5 heterodimers as a novel cell surface marker of human pancreatic beta cells, a finding that will aid in the identification and characterisation of this important cell type.
Published: 2021
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6. Enhanced structure and function of human pluripotent stem cell‐derived beta‐cells cultured on extracellular matrix

Author: Reena Singh, Louise Cottle, Thomas Loudovaris, Di Xiao, Pengyi Yang, Helen E. Thomas, Melkam A. Kebede, and Peter Thorn
Subjects: basement membrane, diabetes, differentiation, glucose stimulated insulin secretion, human pluripotent stem cell‐derived beta cells, Medicine (General), R5-920, Cytology, QH573-671
Abstract: Abstract The differentiation of human stem cells into insulin secreting beta‐like cells holds great promise to treat diabetes. Current protocols drive stem cells through stages of directed differentiation and maturation and produce cells that secrete insulin in response to glucose. Further refinements are now needed to faithfully phenocopy the responses of normal beta cells. A critical factor in normal beta cell behavior is the islet microenvironment which plays a central role in beta cell survival, proliferation, gene expression and secretion. One important influence on native cell responses is the capillary basement membrane. In adult islets, each beta cell makes a point of contact with basement membrane protein secreted by vascular endothelial cells resulting in structural and functional polarization. Interaction with basement membrane proteins triggers local activation of focal adhesions, cell orientation, and targeting of insulin secretion. This study aims to identifying the role of basement membrane proteins on the structure and function of human embryonic stem cell and induced pluripotent stem cell‐derived beta cells. Here, we show that differentiated human stem cells‐derived spheroids do contain basement membrane proteins as a diffuse web‐like structure. However, the beta‐like cells within the spheroid do not polarize in response to this basement membrane. We demonstrate that 2D culture of the differentiated beta cells on to basement membrane proteins enforces cell polarity and favorably alters glucose dependent insulin secretion.
Published: 2021
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7. Case Report: Hypoglycemia Due to a Novel Activating Glucokinase Variant in an Adult – a Molecular Approach

Author: Anojian Koneshamoorthy, Dilan Seniveratne-Epa, Genevieve Calder, Matthew Sawyer, Thomas W. H. Kay, Stephen Farrell, Thomas Loudovaris, Lina Mariana, Davis McCarthy, Ruqian Lyu, Xin Liu, Peter Thorn, Jason Tong, Lit Kim Chin, Margaret Zacharin, Alison Trainer, Shelby Taylor, Richard J. MacIsaac, Nirupa Sachithanandan, Helen E. Thomas, and Balasubramanian Krishnamurthy
Subjects: hypoglycemia, glucokinase, congenital hyperinsulinism, hyperplastic islets, MODY, Diseases of the endocrine glands. Clinical endocrinology, RC648-665
Abstract: We present a case of an obese 22-year-old man with activating GCK variant who had neonatal hypoglycemia, re-emerging with hypoglycemia later in life. We investigated him for asymptomatic hypoglycemia with a family history of hypoglycemia. Genetic testing yielded a novel GCK missense class 3 variant that was subsequently found in his mother, sister and nephew and reclassified as a class 4 likely pathogenic variant. Glucokinase enables phosphorylation of glucose, the rate-limiting step of glycolysis in the liver and pancreatic β cells. It plays a crucial role in the regulation of insulin secretion. Inactivating variants in GCK cause hyperglycemia and activating variants cause hypoglycemia. Spleen-preserving distal pancreatectomy revealed diffuse hyperplastic islets, nuclear pleomorphism and periductular islets. Glucose stimulated insulin secretion revealed increased insulin secretion in response to glucose. Cytoplasmic calcium, which triggers exocytosis of insulin-containing granules, revealed normal basal but increased glucose-stimulated level. Unbiased gene expression analysis using 10X single cell sequencing revealed upregulated INS and CKB genes and downregulated DLK1 and NPY genes in β-cells. Further studies are required to see if alteration in expression of these genes plays a role in the metabolic and histological phenotype associated with glucokinase pathogenic variant. There were more large islets in the patient’s pancreas than in control subjects but there was no difference in the proportion of β cells in the islets. His hypoglycemia was persistent after pancreatectomy, was refractory to diazoxide and improved with pasireotide. This case highlights the variable phenotype of GCK mutations. In-depth molecular analyses in the islets have revealed possible mechanisms for hyperplastic islets and insulin hypersecretion.
Published: 2022
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8. Circulating platelet-neutrophil aggregates characterize the development of type 1 diabetes in humans and NOD mice

Author: Sarah K. Popp, Federica Vecchio, Debra J. Brown, Riho Fukuda, Yuri Suzuki, Yuma Takeda, Rikako Wakamatsu, Mahalakshmi A. Sarma, Jessica Garrett, Anna Giovenzana, Emanuele Bosi, Antony R.A. Lafferty, Karen J. Brown, Elizabeth E. Gardiner, Lucy A. Coupland, Helen E. Thomas, Beng H. Chong, Christopher R. Parish, Manuela Battaglia, Alessandra Petrelli, and Charmaine J. Simeonovic
Subjects: Autoimmunity, Medicine
Abstract: Platelet-neutrophil aggregates (PNAs) facilitate neutrophil activation and migration and could underpin the recruitment of neutrophils to the pancreas during type 1 diabetes (T1D) pathogenesis. PNAs, measured by flow cytometry, were significantly elevated in the circulation of autoantibody-positive (Aab+) children and new-onset T1D children, as well as in pre-T1D (at 4 weeks and 10–12 weeks) and T1D-onset NOD mice, compared with relevant controls, and PNAs were characterized by activated P-selectin+ platelets. PNAs were similarly increased in pre-T1D and T1D-onset NOD isolated islets/insulitis, and immunofluorescence staining revealed increased islet-associated neutrophil extracellular trap (NET) products (myeloperoxidase [MPO] and citrullinated histones [CitH3]) in NOD pancreata. In vitro, cell-free histones and NETs induced islet cell damage, which was prevented by the small polyanionic drug methyl cellobiose sulfate (mCBS) that binds to histones and neutralizes their pathological effects. Elevated circulating PNAs could, therefore, act as an innate immune and pathogenic biomarker of T1D autoimmunity. Platelet hyperreactivity within PNAs appears to represent a previously unrecognized hematological abnormality that precedes T1D onset. In summary, PNAs could contribute to the pathogenesis of T1D and potentially function as a pre-T1D diagnostic.
Published: 2022
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9. Neuropeptide Y1 receptor antagonism protects β-cells and improves glycemic control in type 2 diabetes

Author: Chieh-Hsin Yang, Danise Ann-Onda, Xuzhu Lin, Stacey Fynch, Shaktypreya Nadarajah, Evan G. Pappas, Xin Liu, John W. Scott, Jonathan S. Oakhill, Sandra Galic, Yanchuan Shi, Alba Moreno-Asso, Cassandra Smith, Thomas Loudovaris, Itamar Levinger, Decio L. Eizirik, D. Ross Laybutt, Herbert Herzog, Helen E. Thomas, and Kim Loh
Subjects: NPY, β-Cell, Insulin secretion, Y1 receptor, Type 2 diabetes, Internal medicine, RC31-1245
Abstract: Objectives: Loss of functional β-cell mass is a key factor contributing to poor glycemic control in advanced type 2 diabetes (T2D). We have previously reported that the inhibition of the neuropeptide Y1 receptor improves the islet transplantation outcome in type 1 diabetes (T1D). The aim of this study was to identify the pathophysiological role of the neuropeptide Y (NPY) system in human T2D and further evaluate the therapeutic potential of using the Y1 receptor antagonist BIBO3304 to improve β-cell function and survival in T2D. Methods: The gene expression of the NPY system in human islets from nondiabetic subjects and subjects with T2D was determined and correlated with the stimulation index. The glucose-lowering and β-cell-protective effects of BIBO3304, a selective orally bioavailable Y1 receptor antagonist, in high-fat diet (HFD)/multiple low-dose streptozotocin (STZ)-induced and genetically obese (db/db) T2D mouse models were assessed. Results: In this study, we identified a more than 2-fold increase in NPY1R and its ligand, NPY mRNA expression in human islets from subjects with T2D, which was significantly associated with reduced insulin secretion. Consistently, the pharmacological inhibition of Y1 receptors by BIBO3304 significantly protected β cells from dysfunction and death under multiple diabetogenic conditions in islets. In a preclinical study, we demonstrated that the inhibition of Y1 receptors by BIBO3304 led to reduced adiposity and enhanced insulin action in the skeletal muscle. Importantly, the Y1 receptor antagonist BIBO3304 treatment also improved β-cell function and preserved functional β-cell mass, thereby resulting in better glycemic control in both HFD/multiple low-dose STZ-induced and db/db T2D mice. Conclusions: Our results revealed a novel causal link between increased islet NPY-Y1 receptor gene expression and β-cell dysfunction and failure in human T2D, contributing to the understanding of the pathophysiology of T2D. Furthermore, our results demonstrate that the inhibition of the Y1 receptor by BIBO3304 represents a potential β-cell-protective therapy for improving functional β-cell mass and glycemic control in T2D.
Published: 2022
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10. A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial CellsSummary

Author: Mugdha V. Joglekar, Subhshri Sahu, Wilson K.M. Wong, Sarang N. Satoor, Charlotte X. Dong, Ryan J. Farr, Michael D. Williams, Prapti Pandya, Gaurang Jhala, Sundy N.Y. Yang, Yi Vee Chew, Nicola Hetherington, Dhan Thiruchevlam, Sasikala Mitnala, Guduru V. Rao, Duvvuru Nageshwar Reddy, Thomas Loudovaris, Wayne J. Hawthorne, Andrew G. Elefanty, Vinay M. Joglekar, Edouard G. Stanley, David Martin, Helen E. Thomas, David Tosh, Louise T. Dalgaard, and Anandwardhan A. Hardikar
Subjects: Islets, Insulin, Splice Variants, Gallbladder Development, Differentiation, Diseases of the digestive system. Gastroenterology, RC799-869
Abstract: Background & Aims: Pancreatic islet β-cells are factories for insulin production; however, ectopic expression of insulin also is well recognized. The gallbladder is a next-door neighbor to the developing pancreas. Here, we wanted to understand if gallbladders contain functional insulin-producing cells. Methods: We compared developing and adult mouse as well as human gallbladder epithelial cells and islets using immunohistochemistry, flow cytometry, enzyme-linked immunosorbent assays, RNA sequencing, real-time polymerase chain reaction, chromatin immunoprecipitation, and functional studies. Results: We show that the epithelial lining of developing, as well as adult, mouse and human gallbladders naturally contain interspersed cells that retain the capacity to actively transcribe, translate, package, and release insulin. We show that human gallbladders also contain functional insulin-secreting cells with the potential to naturally respond to glucose in vitro and in situ. Notably, in a non-obese diabetic (NOD) mouse model of type 1 diabetes, we observed that insulin-producing cells in the gallbladder are not targeted by autoimmune cells. Interestingly, in human gallbladders, insulin splice variants are absent, although insulin splice forms are observed in human islets. Conclusions: In summary, our biochemical, transcriptomic, and functional data in mouse and human gallbladder epithelial cells collectively show the evolutionary and developmental similarities between gallbladder and the pancreas that allow gallbladder epithelial cells to continue insulin production in adult life. Understanding the mechanisms regulating insulin transcription and translation in gallbladder epithelial cells would help guide future studies in type 1 diabetes therapy.
Published: 2022
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11. Machine learning workflows identify a microRNA signature of insulin transcription in human tissues

Author: Wilson K.M. Wong, Mugdha V. Joglekar, Vijit Saini, Guozhi Jiang, Charlotte X. Dong, Alissa Chaitarvornkit, Grzegorz J. Maciag, Dario Gerace, Ryan J. Farr, Sarang N. Satoor, Subhshri Sahu, Tejaswini Sharangdhar, Asma S. Ahmed, Yi Vee Chew, David Liuwantara, Benjamin Heng, Chai K. Lim, Julie Hunter, Andrzej S. Januszewski, Anja E. Sørensen, Ammira S.A. Akil, Jennifer R. Gamble, Thomas Loudovaris, Thomas W. Kay, Helen E. Thomas, Philip J. O'Connell, Gilles J. Guillemin, David Martin, Ann M. Simpson, Wayne J. Hawthorne, Louise T. Dalgaard, Ronald C.W. Ma, and Anandwardhan A. Hardikar
Subjects: Pathophysiology, Computational Bioinformatics, Transcriptomics, Science
Abstract: Summary: Dicer knockout mouse models demonstrated a key role for microRNAs in pancreatic β-cell function. Studies to identify specific microRNA(s) associated with human (pro-)endocrine gene expression are needed. We profiled microRNAs and key pancreatic genes in 353 human tissue samples. Machine learning workflows identified microRNAs associated with (pro-)insulin transcripts in a discovery set of islets (n = 30) and insulin-negative tissues (n = 62). This microRNA signature was validated in remaining 261 tissues that include nine islet samples from individuals with type 2 diabetes. Top eight microRNAs (miR-183-5p, -375-3p, 216b-5p, 183-3p, -7-5p, -217-5p, -7-2-3p, and -429-3p) were confirmed to be associated with and predictive of (pro-)insulin transcript levels. Use of doxycycline-inducible microRNA-overexpressing human pancreatic duct cell lines confirmed the regulatory roles of these microRNAs in (pro-)endocrine gene expression. Knockdown of these microRNAs in human islet cells reduced (pro-)insulin transcript abundance. Our data provide specific microRNAs to further study microRNA-mRNA interactions in regulating insulin transcription.
Published: 2021
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12. Tolerance to Proinsulin-1 Reduces Autoimmune Diabetes in NOD Mice

Author: Gaurang Jhala, Claudia Selck, Jonathan Chee, Chun-Ting J. Kwong, Evan G. Pappas, Helen E. Thomas, Thomas W.H. Kay, and Balasubramanian Krishnamurthy
Subjects: type 1 diabetes, proinsulin-1, CD4+ T cells, immune tolerance, NOD mice, Immunologic diseases. Allergy, RC581-607
Abstract: T-cell responses to insulin and its precursor proinsulin are central to islet autoimmunity in humans and non-obese diabetic (NOD) mice that spontaneously develop autoimmune diabetes. Mice have two proinsulin genes proinsulin -1 and 2 that are differentially expressed, with predominant proinsulin-2 expression in the thymus and proinsulin-1 in islet beta-cells. In contrast to proinsulin-2, proinsulin-1 knockout NOD mice are protected from autoimmune diabetes. This indicates that proinsulin-1 epitopes in beta-cells maybe preferentially targeted by autoreactive T cells. To study the contribution of proinsulin-1 reactive T cells in autoimmune diabetes, we generated transgenic NOD mice with tetracycline-regulated expression of proinsulin-1 in antigen presenting cells (TIP-1 mice) with an aim to induce immune tolerance. TIP-1 mice displayed a significantly reduced incidence of spontaneous diabetes, which was associated with reduced severity of insulitis and insulin autoantibody development. Antigen experienced proinsulin specific T cells were significantly reduced in in TIP-1 mice indicating immune tolerance. Moreover, T cells from TIP-1 mice expressing proinsulin-1 transferred diabetes at a significantly reduced frequency. However, proinsulin-1 expression in APCs had minimal impact on the immune responses to the downstream antigen islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) and did not prevent diabetes in NOD 8.3 mice with a pre-existing repertoire of IGRP reactive T cells. Thus, boosting immune tolerance to proinsulin-1 partially prevents islet-autoimmunity. This study further extends the previously established role of proinsulin-1 epitopes in autoimmune diabetes in NOD mice.
Published: 2021
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13. The JAK1 Selective Inhibitor ABT 317 Blocks Signaling Through Interferon-γ and Common γ Chain Cytokine Receptors to Reverse Autoimmune Diabetes in NOD Mice

Author: Tingting Ge, Gaurang Jhala, Stacey Fynch, Satoru Akazawa, Sara Litwak, Evan G. Pappas, Tara Catterall, Ishan Vakil, Andrew J. Long, Lisa M. Olson, Balasubramanian Krishnamurthy, Thomas W. Kay, and Helen E. Thomas
Subjects: JAK-STAT signaling pathway, cytokines, non-obese diabetic mouse, CD8+ T cell, type 1 diabetes mellitus, Immunologic diseases. Allergy, RC581-607
Abstract: Cytokines that signal through the JAK-STAT pathway, such as interferon-γ (IFN-γ) and common γ chain cytokines, contribute to the destruction of insulin-secreting β cells by CD8+ T cells in type 1 diabetes (T1D). We previously showed that JAK1/JAK2 inhibitors reversed autoimmune insulitis in non-obese diabetic (NOD) mice and also blocked IFN-γ mediated MHC class I upregulation on β cells. Blocking interferons on their own does not prevent diabetes in knockout NOD mice, so we tested whether JAK inhibitor action on signaling downstream of common γ chain cytokines, including IL-2, IL-7 IL-15, and IL-21, may also affect the progression of diabetes in NOD mice. Common γ chain cytokines activate JAK1 and JAK3 to regulate T cell proliferation. We used a JAK1-selective inhibitor, ABT 317, to better understand the specific role of JAK1 signaling in autoimmune diabetes. ABT 317 reduced IL-21, IL-2, IL-15 and IL-7 signaling in T cells and IFN-γ signaling in β cells, but ABT 317 did not affect GM-CSF signaling in granulocytes. When given in vivo to NOD mice, ABT 317 reduced CD8+ T cell proliferation as well as the number of KLRG+ effector and CD44hiCD62Llo effector memory CD8+ T cells in spleen. ABT 317 also prevented MHC class I upregulation on β cells. Newly diagnosed diabetes was reversed in 94% NOD mice treated twice daily with ABT 317 while still on treatment at 40 days and 44% remained normoglycemic after a further 60 days from discontinuing the drug. Our results indicate that ABT 317 blocks common γ chain cytokines in lymphocytes and interferons in lymphocytes and β cells and are thus more effective against diabetes pathogenesis than IFN-γ receptor deficiency alone. Our studies suggest use of this class of drug for the treatment of type 1 diabetes.
Published: 2020
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14. Machine Learning Algorithms, Applied to Intact Islets of Langerhans, Demonstrate Significantly Enhanced Insulin Staining at the Capillary Interface of Human Pancreatic β Cells

Author: Louise Cottle, Ian Gilroy, Kylie Deng, Thomas Loudovaris, Helen E. Thomas, Anthony J. Gill, Jaswinder S. Samra, Melkam A. Kebede, Jinman Kim, and Peter Thorn
Subjects: insulin, beta cell, human, islet, polarisation, machine learning, Microbiology, QR1-502
Abstract: Pancreatic β cells secrete the hormone insulin into the bloodstream and are critical in the control of blood glucose concentrations. β cells are clustered in the micro-organs of the islets of Langerhans, which have a rich capillary network. Recent work has highlighted the intimate spatial connections between β cells and these capillaries, which lead to the targeting of insulin secretion to the region where the β cells contact the capillary basement membrane. In addition, β cells orientate with respect to the capillary contact point and many proteins are differentially distributed at the capillary interface compared with the rest of the cell. Here, we set out to develop an automated image analysis approach to identify individual β cells within intact islets and to determine if the distribution of insulin across the cells was polarised. Our results show that a U-Net machine learning algorithm correctly identified β cells and their orientation with respect to the capillaries. Using this information, we then quantified insulin distribution across the β cells to show enrichment at the capillary interface. We conclude that machine learning is a useful analytical tool to interrogate large image datasets and analyse sub-cellular organisation.
Published: 2021
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15. Inhibition of Y1 receptor signaling improves islet transplant outcome

Author: Kim Loh, Yan-Chuan Shi, Stacey Walters, Mohammed Bensellam, Kailun Lee, Katsuya Dezaki, Masanori Nakata, Chi Kin Ip, Jeng Yie Chan, Esteban N. Gurzov, Helen E. Thomas, Michaela Waibel, James Cantley, Thomas W. Kay, Toshihiko Yada, D. Ross Laybutt, Shane T. Grey, and Herbert Herzog
Subjects: Science
Abstract: Islet transplantation is considered one of the potential treatments for T1DM but limited islet survival and their impaired function pose limitations to this approach. Here Loh et al. show that the Y1 receptor is expressed in β- cells and inhibition of its signalling, both genetic and pharmacological, improves mouse and human islet function.
Published: 2017
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16. IFNγ-Induced MHC Class II Expression on Islet Endothelial Cells Is an Early Marker of Insulitis but Is Not Required for Diabetogenic CD4+ T Cell Migration

Author: Nicholas A. Scott, Yuxing Zhao, Balasubramanian Krishnamurthy, Stuart I. Mannering, Thomas W. H. Kay, and Helen E. Thomas
Subjects: Type 1 diabetes, MHC class II, interferon-γ, endothelial cells, non-obese diabetic mice, Immunologic diseases. Allergy, RC581-607
Abstract: Diabetogenic T cells infiltrate the pancreatic islets by transmigrating across the microcapillaries residing close to, or within, the pancreatic islets. Deficiency in IFNγ signaling prevents efficient migration of T cells into the pancreatic islets, but the IFNγ-regulated molecules that mediate this are uncertain. Homing of autoreactive T cells into target tissues may require antigen specificity through presentation of cognate antigen by MHC expressed on the vascular endothelium. We investigated the hypothesis that IFNγ promotes the migration of islet antigen-specific CD4+ T cells by upregulating MHC class II on islet endothelial cells (IEC), thereby providing an antigen-specific signal for islet infiltration. Upon IFNγ stimulation, MHC class II, which is not constitutively expressed on IEC, was induced. IFNγ-dependent upregulation of MHC class II was detected in IEC isolated from prediabetic NOD mice at the earliest stages of insulitis, before other markers of inflammation were present. Using a CD4+ T cell-mediated adoptive transfer model of autoimmune diabetes we observed that even though diabetes does not develop in recipient mice lacking IFNγ receptors, mice with MHC class II-deficient IEC were not protected from disease. Thus, IFNγ-regulated molecules, but not MHC class II or antigen presentation by IECs is required for the early migration of antigen-specific CD4+ T cells into the pancreatic islets.
Published: 2018
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17. Linking Metabolic Abnormalities to Apoptotic Pathways in Beta Cells in Type 2 Diabetes

Author: Helen E. Thomas, Seth L. Masters, and Jibran A. Wali
Subjects: Type 2 diabetes, pancreatic beta cell, apoptosis, endoplasmic reticulum stress, oxidative stress, NLRP3 inflammasome, Bcl-2 pathway, Cytology, QH573-671
Abstract: Pancreatic beta-cell apoptosis is an important feature of islets in type 2 diabetes. Apoptosis can occur through two major pathways, the extrinsic or death receptor mediated pathway, and the intrinsic or Bcl-2-regulated pathway. Hyperglycaemia, hyperlipidaemia and islet amyloid poly-peptide (IAPP) represent important possible causes of increased beta-cell apoptosis. Hyperglycaemia induces islet-cell apoptosis by the intrinsic pathway involving molecules of the Bcl-2 family. High concentrations of palmitate also activate intrinsic apoptosis in islets cells. IAPP oligomers can induce apoptosis by both intrinsic and extrinsic pathways. IL-1b produced through NLRP3 inflammasome activation can also induce islet cell death. Activation of the NLRP3 inflammasome may not be important for glucose or palmitate induced apoptosis in islets but may be important for IAPP mediated cell death. Endoplasmic reticulum (ER) and oxidative stress have been observed in beta cells in type 2 diabetes, and these could be the link between upstream metabolic abnormalities and downstream apoptotic machinery.
Published: 2013
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18. Type I Interferon Signaling Is Required for Dacryoadenitis in the Nonobese Diabetic Mouse Model of Sjögren Syndrome

Author: Yury Chaly, Jennifer Y. Barr, David A. Sullivan, Helen E. Thomas, Thomas C. Brodnicki, and Scott M. Lieberman
Subjects: Sjögren syndrome, lacrimal glands, nonobese diabetic mice, regulatory T cells, type I interferon, chemokines, Biology (General), QH301-705.5, Chemistry, QD1-999
Abstract: Nonobese diabetic (NOD) mice spontaneously develop lacrimal and salivary gland autoimmunity similar to human Sjögren syndrome. In both humans and NOD mice, the early immune response that drives T-cell infiltration into lacrimal and salivary glands is poorly understood. In NOD mice, lacrimal gland autoimmunity spontaneously occurs only in males with testosterone playing a role in promoting lacrimal gland inflammation, while female lacrimal glands are protected by regulatory T cells (Tregs). The mechanisms of this male-specific lacrimal gland autoimmunity are not known. Here, we studied the effects of Treg depletion in hormone-manipulated NOD mice and lacrimal gland gene expression to determine early signals required for lacrimal gland inflammation. While Treg-depletion was not sufficient to drive dacryoadenitis in castrated male NOD mice, chemokines (Cxcl9, Ccl19) and other potentially disease-relevant genes (Epsti1, Ubd) were upregulated in male lacrimal glands. Expression of Cxcl9 and Ccl19, in particular, remained significantly upregulated in the lacrimal glands of lymphocyte-deficient NOD-severe combined immunodeficiency (SCID) mice and their expression was modulated by type I interferon signaling. Notably, Ifnar1-deficient NOD mice did not develop dacryoadenitis. Together these data identify disease-relevant genes upregulated in the context of male-specific dacryoadenitis and demonstrate a requisite role for type I interferon signaling in lacrimal gland autoimmunity in NOD mice.
Published: 2018
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19. Lipotoxic Stress Induces Pancreatic β-Cell Apoptosis through Modulation of Bcl-2 Proteins by the Ubiquitin-Proteasome System

Author: Sara A. Litwak, Jibran A. Wali, Evan G. Pappas, Hamdi Saadi, William J. Stanley, L. Chitra Varanasi, Thomas W. H. Kay, Helen E. Thomas, and Esteban N. Gurzov
Subjects: Diseases of the endocrine glands. Clinical endocrinology, RC648-665
Abstract: Pancreatic β-cell loss induced by saturated free fatty acids (FFAs) is believed to contribute to type 2 diabetes. Previous studies have shown induction of endoplasmic reticulum (ER) stress, increased ubiquitinated proteins, and deregulation of the Bcl-2 family in the pancreas of type 2 diabetic patients. However, the precise mechanism of β-cell death remains unknown. In the present study we demonstrate that the FFA palmitate blocks the ubiquitin-proteasome system (UPS) and causes apoptosis through induction of ER stress and deregulation of Bcl-2 proteins. We found that palmitate and the proteasome inhibitor MG132 induced ER stress in β-cells, resulting in decreased expression of the prosurvival proteins Bcl-2, Mcl-1, and Bcl-XL, and upregulation of the prodeath BH3-only protein PUMA. On the other hand, pharmacological activation of the UPS by sulforaphane ameliorated ER stress, upregulated prosurvival Bcl-2 proteins, and protected β-cells from FFA-induced cell death. Furthermore, transgenic overexpression of Bcl-2 protected islets from FFA-induced cell death in vitro and improved glucose-induced insulin secretion in vivo. Together our results suggest that targeting the UPS and Bcl-2 protein expression may be a valuable strategy to prevent β-cell demise in type 2 diabetes.
Published: 2015
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20. Pancreatic Alpha Cells Hold the Key to Survival

Author: Jibran A. Wali and Helen E. Thomas
Subjects: Medicine, Medicine (General), R5-920
Published: 2015
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21. Hypoxia-Inducible Factor-1α (HIF-1α) Potentiates β-Cell Survival after Islet Transplantation of Human and Mouse Islets

Author: Rebecca A. Stokes, Kim Cheng, Natasha Deters, Sue Mei Lau, Wayne J. Hawthorne, Philip J. O'connell, Jessica Stolp, Shane Grey, Thomas Loudovaris, Thomas W. Kay, Helen E. Thomas, Frank J. Gonzalez, and Jenny E. Gunton
Subjects: Medicine
Abstract: A high proportion of β-cells die within days of islet transplantation. Reports suggest that induction of hypoxia-inducible factor-1α (HIF-1α) predicts adverse transplant outcomes. We hypothesized that this was a compensatory response and that HIF-1α protects β-cells during transplantation. Transplants were performed using human islets or murine β-cell-specific HIF-1α-null (β-HIF-1α-null) islets with or without treatment with deferoxamine (DFO) to increase HIF-1α. β-HIF-1α-null transplants had poor outcomes, demonstrating that lack of HIF-1α impaired transplant efficiency. Increasing HIF-1α improved outcomes for mouse and human islets. No effect was seen in β-HIF-1α-null islets. The mechanism was decreased apoptosis, resulting in increased β-cell mass posttransplantation. These findings show that HIF-1α is a protective factor and is required for successful islet transplant outcomes. Iron chelation with DFO markedly improved transplant success in a HIF-1α-dependent manner, thus demonstrating the mechanism of action. DFO, approved for human use, may have a therapeutic role in the setting of human islet transplantation.
Published: 2013
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22. Expression of Pro- and Antiapoptotic Molecules of the Bcl-2 Family in Human Islets Postisolation

Author: Peter D. Campbell, Anita Weinberg, Jonathan Chee, Lina Mariana, Rochelle Ayala, Wayne J. Hawthorne, Philip J. O'connell, Thomas Loudovaris, Mark J. Cowley, Thomas W. Kay, Shane T. Grey, and Helen E. Thomas
Subjects: Medicine
Abstract: Human islets are subjected to a number of stresses before and during their isolation that may influence their survival and engraftment after transplantation. Apoptosis is likely to be activated in response to these stresses. Apoptosis due to intrinsic stresses is regulated by pro- and antiapoptotic members of the Bcl-2 family. While the role of the Bcl-2 family in apoptosis of rodent islets is becoming increasingly understood, little is known about which of these molecules are expressed or required for apoptosis of human islets. This study investigated the expression of the Bcl-2 family of molecules in isolated human islets. RNA and protein lysates were extracted from human islets immediately postisolation. At the same time, standard quality control assays including viability staining and β-cell content were performed on each islet preparation. Microarrays, RT-PCR, and Western blotting were performed on islet RNA and protein. The prosurvival molecules Bcl-xl and Mcl-1, but not Bcl-2, were highly expressed. The multidomain proapoptotic effector molecule Bax was expressed at higher levels than Bak. Proapoptotic BH3-only molecules were expressed at low levels, with Bid being the most abundant. The proapoptotic molecules BNIP3, BNIP3L, and Beclin-1 were all highly expressed, indicating exposure of islets to oxygen and nutrient deprivation during isolation. Our data provide a comprehensive analysis of expression levels of pro- and antiapoptotic Bcl-2 family members in isolated human islets. Knowledge of which molecules are expressed will guide future research to understand the apoptotic pathways activated during isolation or after transplantation. This is crucial for the design of methods to achieve improved transplantation outcomes.
Published: 2012
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23. Enhanced structure and function of human pluripotent stem cell‐derived beta‐cells cultured on extracellular matrix

Author: Thomas Loudovaris, Pengyi Yang, Melkam A. Kebede, Reena Singh, Helen E. Thomas, Louise Cottle, Di Xiao, and Peter Thorn
Subjects: 0301 basic medicine, Pluripotent Stem Cells, 03 medical and health sciences, 0302 clinical medicine, Directed differentiation, Tissue Engineering and Regenerative Medicine, human pluripotent stem cell‐derived beta cells, Insulin-Secreting Cells, Cell polarity, medicine, Humans, Insulin, Secretion, lcsh:QH573-671, Induced pluripotent stem cell, Basement membrane, lcsh:R5-920, diabetes, lcsh:Cytology, Chemistry, Endothelial Cells, Cell Differentiation, Cell Biology, General Medicine, differentiation, Embryonic stem cell, basement membrane, Cell biology, Extracellular Matrix, 030104 developmental biology, medicine.anatomical_structure, Glucose, glucose stimulated insulin secretion, Beta cell, Stem cell, lcsh:Medicine (General), 030217 neurology & neurosurgery, Developmental Biology
Abstract: The differentiation of human stem cells into insulin secreting beta‐like cells holds great promise to treat diabetes. Current protocols drive stem cells through stages of directed differentiation and maturation and produce cells that secrete insulin in response to glucose. Further refinements are now needed to faithfully phenocopy the responses of normal beta cells. A critical factor in normal beta cell behavior is the islet microenvironment which plays a central role in beta cell survival, proliferation, gene expression and secretion. One important influence on native cell responses is the capillary basement membrane. In adult islets, each beta cell makes a point of contact with basement membrane protein secreted by vascular endothelial cells resulting in structural and functional polarization. Interaction with basement membrane proteins triggers local activation of focal adhesions, cell orientation, and targeting of insulin secretion. This study aims to identifying the role of basement membrane proteins on the structure and function of human embryonic stem cell and induced pluripotent stem cell‐derived beta cells. Here, we show that differentiated human stem cells‐derived spheroids do contain basement membrane proteins as a diffuse web‐like structure. However, the beta‐like cells within the spheroid do not polarize in response to this basement membrane. We demonstrate that 2D culture of the differentiated beta cells on to basement membrane proteins enforces cell polarity and favorably alters glucose dependent insulin secretion., Glucose stimulated insulin secretion is compromised in stem cell‐derived beta cells. In native islets, beta cell microenvironment play critical role in glucose sensing and stimulated insulin secretion. Reconstructing beta cell niche in vitro by introducing basement membrane proteins in planar culture, decreases basal insulin secretion, and enhances stimulation index in human pluripotent stem cell‐derived beta cells.
Published: 2020

24. Machine learning workflows identify a microRNA signature of insulin transcription in human tissues

Author: Philip J. O'Connell, Thomas Loudovaris, Mugdha V. Joglekar, Anja Elaine Sørensen, Ann M. Simpson, Helen E. Thomas, Louise Torp Dalgaard, Ryan J. Farr, Ronald C.W. Ma, Tejaswini Sharangdhar, Ammira S. Akil, Benjamin Heng, Julie Hunter, David Martín, Yi Vee Chew, Charlotte X. Dong, Sarang N. Satoor, Guozhi Jiang, Alissa Chaitarvornkit, David Liuwantara, Andrzej S. Januszewski, Dario Gerace, Jennifer R. Gamble, Vijit Saini, Anandwardhan A. Hardikar, Wayne J. Hawthorne, Chai K. Lim, Asma S. Ahmed, Subhshri Sahu, Wilson K. M. Wong, Gilles J. Guillemin, Grzegorz J. Maciag, and Thomas W.H. Kay
Subjects: 0301 basic medicine, medicine.medical_treatment, Science, Computational Bioinformatics, 02 engineering and technology, Machine learning, computer.software_genre, Pathophysiology, Article, Transcriptome, 03 medical and health sciences, Gene expression, microRNA, medicine, Transcriptomics, Gene, Gene knockdown, Multidisciplinary, biology, business.industry, Insulin, 021001 nanoscience & nanotechnology, 030104 developmental biology, Knockout mouse, biology.protein, Artificial intelligence, 0210 nano-technology, business, computer, Dicer
Abstract: Summary Dicer knockout mouse models demonstrated a key role for microRNAs in pancreatic β-cell function. Studies to identify specific microRNA(s) associated with human (pro-)endocrine gene expression are needed. We profiled microRNAs and key pancreatic genes in 353 human tissue samples. Machine learning workflows identified microRNAs associated with (pro-)insulin transcripts in a discovery set of islets (n = 30) and insulin-negative tissues (n = 62). This microRNA signature was validated in remaining 261 tissues that include nine islet samples from individuals with type 2 diabetes. Top eight microRNAs (miR-183-5p, -375-3p, 216b-5p, 183-3p, -7-5p, -217-5p, -7-2-3p, and -429-3p) were confirmed to be associated with and predictive of (pro-)insulin transcript levels. Use of doxycycline-inducible microRNA-overexpressing human pancreatic duct cell lines confirmed the regulatory roles of these microRNAs in (pro-)endocrine gene expression. Knockdown of these microRNAs in human islet cells reduced (pro-)insulin transcript abundance. Our data provide specific microRNAs to further study microRNA-mRNA interactions in regulating insulin transcription., Graphical abstract, Highlights • Unbiased machine learning workflow ranks miRNAs associated with insulin transcription • Forced expression of top-ranked miRNAs drives pro-endocrine program in progenitor cells • Knockdown of top-ranked miRNAs retards insulin gene transcription in human islets • Insulin transcript-associated miRNAs are reduced in islets of donors with type 2 diabetes, Pathophysiology; Computational Bioinformatics; Transcriptomics
Published: 2021

25. ABCA12 regulates insulin secretion from β‐cells

Author: Ying Fu, Helena C. Parkington, Nigora Mukhamedova, Gloria M. A. Ursino, Natalie A. Mellett, Lynelle K. Jones, Stacey Fynch, Peter J. Meikle, Dmitri Sviridov, Hann Low, Peter Thorn, Jacquelyn M. Weir, Michael Ditiatkovski, Denny L Cottle, Wan Jun Gan, Helen E. Thomas, Partha Pratim Das, Ming Shen Tham, and Ian M. Smyth
Subjects: Ceramide, medicine.medical_treatment, Inflammation, Biochemistry, Cell membrane, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, Insulin-Secreting Cells, Insulin Secretion, Genetics, medicine, Animals, Insulin, Molecular Biology, Lipid raft, 030304 developmental biology, 0303 health sciences, biology, Chemistry, Articles, Cell biology, medicine.anatomical_structure, Glucose, Diabetes Mellitus, Type 2, ABCA1, biology.protein, lipids (amino acids, peptides, and proteins), ATP-Binding Cassette Transporters, medicine.symptom, Pancreas, 030217 neurology & neurosurgery, Homeostasis, ATP Binding Cassette Transporter 1
Abstract: Dysregulation of lipid homeostasis is intimately associated with defects in insulin secretion, a key feature of type 2 diabetes. Here, we explore the role of the putative lipid transporter ABCA12 in regulating insulin secretion from β-cells. Mice with β-cell-specific deletion of Abca12 display impaired glucose-stimulated insulin secretion and eventual islet inflammation and β-cell death. ABCA12's action in the pancreas is independent of changes in the abundance of two other cholesterol transporters, ABCA1 and ABCG1, or of changes in cellular cholesterol or ceramide content. Instead, loss of ABCA12 results in defects in the genesis and fusion of insulin secretory granules and increases in the abundance of lipid rafts at the cell membrane. These changes are associated with dysregulation of the small GTPase CDC42 and with decreased actin polymerisation. Our findings establish a new, pleiotropic role for ABCA12 in regulating pancreatic lipid homeostasis and insulin secretion.
Published: 2020

26. Inhibition of Y1 receptor signaling improves islet transplant outcome

Author: Jeng Yie Chan, Thomas W.H. Kay, Katsuya Dezaki, Kim Loh, James Cantley, Esteban Nicolas Gurzov, Stacey N. Walters, Kailun Lee, Chi Kin Ip, Shane St Grey, Ross Laybutt, Toshihiko Yada, Herbert Herzog, Yan-Chuan Shi, Helen E. Thomas, Michaela Waibel, Masanori Nakata, and Mohammed Bensellam
Subjects: 0301 basic medicine, medicine.medical_specialty, endocrine system, endocrine system diseases, medicine.medical_treatment, Science, Islets of Langerhans Transplantation, General Physics and Astronomy, Biology, Arginine, General Biochemistry, Genetics and Molecular Biology, Article, Diabetes Mellitus, Experimental, 03 medical and health sciences, Islets of Langerhans, Mice, Internal medicine, Insulin-Secreting Cells, Insulin Secretion, medicine, Cyclic AMP, Animals, Humans, Insulin, Receptor, lcsh:Science, Islet cell transplantation, geography, Type 1 diabetes, Multidisciplinary, geography.geographical_feature_category, Pancreatic islets, General Chemistry, Sciences bio-médicales et agricoles, Islet, medicine.disease, 3. Good health, Receptors, Neuropeptide Y, Transplantation, 030104 developmental biology, Endocrinology, medicine.anatomical_structure, Cancer research, lcsh:Q, Signal transduction, Signal Transduction
Abstract: Failure to secrete sufficient quantities of insulin is a pathological feature of type-1 and type-2 diabetes, and also reduces the success of islet cell transplantation. Here we demonstrate that Y1 receptor signaling inhibits insulin release in β-cells, and show that this can be pharmacologically exploited to boost insulin secretion. Transplanting islets with Y1 receptor deficiency accelerates the normalization of hyperglycemia in chemically induced diabetic recipient mice, which can also be achieved by short-term pharmacological blockade of Y1 receptors in transplanted mouse and human islets. Furthermore, treatment of non-obese diabetic mice with a Y1 receptor antagonist delays the onset of diabetes. Mechanistically, Y1 receptor signaling inhibits the production of cAMP in islets, which via CREB mediated pathways results in the down-regulation of several key enzymes in glycolysis and ATP production. Thus, manipulating Y1 receptor signaling in β-cells offers a unique therapeutic opportunity for correcting insulin deficiency as it occurs in the pathological state of type-1 diabetes as well as during islet transplantation.Islet transplantation is considered one of the potential treatments for T1DM but limited islet survival and their impaired function pose limitations to this approach. Here Loh et al. show that the Y1 receptor is expressed in β- cells and inhibition of its signalling, both genetic and pharmacological, improves mouse and human islet function., info:eu-repo/semantics/published
Published: 2017

27. Type I Interferon Signaling Is Required for Dacryoadenitis in the Nonobese Diabetic Mouse Model of Sjögren Syndrome

Author: Helen E. Thomas, Thomas C. Brodnicki, David A. Sullivan, Scott M. Lieberman, Jennifer Y. Barr, and Yury Chaly
Subjects: Male, 0301 basic medicine, chemokines, Mice, SCID, Nod, Chemokine CXCL9, T-Lymphocytes, Regulatory, regulatory T cells, lcsh:Chemistry, Mice, 0302 clinical medicine, Mice, Inbred NOD, lcsh:QH301-705.5, Cells, Cultured, Spectroscopy, NOD mice, Salivary gland, Lacrimal Apparatus, Dacryoadenitis, General Medicine, nonobese diabetic mice, Computer Science Applications, Sjogren's Syndrome, medicine.anatomical_structure, Interferon Type I, type I interferon, Female, Signal Transduction, medicine.drug, Lacrimal gland, Lacrimal apparatus, Article, Catalysis, Inorganic Chemistry, Dacryocystitis, 03 medical and health sciences, stomatognathic system, medicine, Animals, Physical and Theoretical Chemistry, Molecular Biology, Autoimmune disease, Organic Chemistry, medicine.disease, Sjögren syndrome, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, Immunology, 030221 ophthalmology & optometry, Chemokine CCL19, Interferon type I, lacrimal glands
Abstract: Nonobese diabetic (NOD) mice spontaneously develop lacrimal and salivary gland autoimmunity similar to human Sj&ouml, gren syndrome. In both humans and NOD mice, the early immune response that drives T-cell infiltration into lacrimal and salivary glands is poorly understood. In NOD mice, lacrimal gland autoimmunity spontaneously occurs only in males with testosterone playing a role in promoting lacrimal gland inflammation, while female lacrimal glands are protected by regulatory T cells (Tregs). The mechanisms of this male-specific lacrimal gland autoimmunity are not known. Here, we studied the effects of Treg depletion in hormone-manipulated NOD mice and lacrimal gland gene expression to determine early signals required for lacrimal gland inflammation. While Treg-depletion was not sufficient to drive dacryoadenitis in castrated male NOD mice, chemokines (Cxcl9, Ccl19) and other potentially disease-relevant genes (Epsti1, Ubd) were upregulated in male lacrimal glands. Expression of Cxcl9 and Ccl19, in particular, remained significantly upregulated in the lacrimal glands of lymphocyte-deficient NOD-severe combined immunodeficiency (SCID) mice and their expression was modulated by type I interferon signaling. Notably, Ifnar1-deficient NOD mice did not develop dacryoadenitis. Together these data identify disease-relevant genes upregulated in the context of male-specific dacryoadenitis and demonstrate a requisite role for type I interferon signaling in lacrimal gland autoimmunity in NOD mice.
Published: 2018
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28. Loss of BIM increases mitochondrial oxygen consumption and lipid oxidation, reduces adiposity and improves insulin sensitivity in mice

Author: David Dr Thorburn, Sean Sl McGee, Bruce E. Kemp, David A. Stroud, Timothy Connor, Thomas W.H. Kay, Chitra Lc Varanasi, Jibran A Wali, Balasubramanian Krishnamurthy, Michael T. Ryan, Evan G Pappas, Ann Ae Frazier, Sandra Galic, Helen E. Thomas, Jingjing Ge, Esteban Nicolas Gurzov, Christina Tan, and Claudia Selck
Subjects: 0301 basic medicine, medicine.medical_specialty, Bioenergetics, Mitochondrion, Carbohydrate metabolism, Biology, Electron Transport Complex IV, Mice, 03 medical and health sciences, Oxygen Consumption, Insulin resistance, Lipid oxidation, Internal medicine, Weight Loss, medicine, Animals, Molecular Biology, Adiposity, Membrane Potential, Mitochondrial, Original Paper, Bcl-2-Like Protein 11, Lipid metabolism, Cell Biology, Metabolism, Lipid Metabolism, medicine.disease, Mitochondria, Glucose, 030104 developmental biology, Endocrinology, Liver, Hepatocytes, Insulin Resistance, biological phenomena, cell phenomena, and immunity, Energy Metabolism, Oxidation-Reduction, Homeostasis
Abstract: BCL-2 proteins are known to engage each other to determine the fate of a cell after a death stimulus. However, their evolutionary conservation and the many other reported binding partners suggest an additional function not directly linked to apoptosis regulation. To identify such a function, we studied mice lacking the BH3-only protein BIM. BIM−/− cells had a higher mitochondrial oxygen consumption rate that was associated with higher mitochondrial complex IV activity. The consequences of increased oxygen consumption in BIM−/− mice were significantly lower body weights, reduced adiposity and lower hepatic lipid content. Consistent with reduced adiposity, BIM−/− mice had lower fasting blood glucose, improved insulin sensitivity and hepatic insulin signalling. Lipid oxidation was increased in BIM−/− mice, suggesting a mechanism for their metabolic phenotype. Our data suggest a role for BIM in regulating mitochondrial bioenergetics and metabolism and support the idea that regulation of metabolism and cell death are connected.
Published: 2018

29. Loss of intra-islet heparan sulfate is a highly sensitive marker of type 1 diabetes progression in humans

Author: Christopher R. Parish, J. Dennis Wilson, Helen E. Thomas, Fui Jiun Choong, Barbara Ludwig, Andrew F. Ziolkowski, Charmaine J. Simeonovic, Sarah K. Popp, Stefan R. Bornstein, Thomas W.H. Kay, Lora Starrs, Craig Freeman, Thomas Loudovaris, Debra J. Brown, Alberto Pugliese, University of Zurich, and Simeonovic, Charmaine J
Subjects: Male, 0301 basic medicine, endocrine system diseases, medicine.medical_treatment, 10265 Clinic for Endocrinology and Diabetology, lcsh:Medicine, Biochemistry, White Blood Cells, chemistry.chemical_compound, Endocrinology, Spectrum Analysis Techniques, 0302 clinical medicine, Animal Cells, Medicine and Health Sciences, Insulin, Child, lcsh:Science, Cells, Cultured, Staining, Multidisciplinary, geography.geographical_feature_category, Chemistry, Cell Staining, Drugs, Heparan sulfate, Flow Cytometry, Islet, 3. Good health, medicine.anatomical_structure, Spectrophotometry, Child, Preschool, Disease Progression, Female, Cytophotometry, Anatomy, Cellular Types, medicine.symptom, Beta cell, Biliary Disorders, Research Article, Adult, medicine.medical_specialty, endocrine system, Adolescent, Endocrine Disorders, Immune Cells, Immunology, Endocrine System, 030209 endocrinology & metabolism, Inflammation, 610 Medicine & health, Gastroenterology and Hepatology, 1100 General Agricultural and Biological Sciences, Research and Analysis Methods, Sensitivity and Specificity, Islets of Langerhans, Young Adult, 03 medical and health sciences, Exocrine Glands, 1300 General Biochemistry, Genetics and Molecular Biology, Internal medicine, Diabetes Mellitus, medicine, Humans, Heparanase, Pancreas, Pharmacology, Diabetic Endocrinology, geography, 1000 Multidisciplinary, Blood Cells, Heparin, Pancreatic islets, lcsh:R, Biology and Life Sciences, Infant, Cell Biology, medicine.disease, Hormones, Diabetes Mellitus, Type 1, 030104 developmental biology, Specimen Preparation and Treatment, Metabolic Disorders, Case-Control Studies, lcsh:Q, Heparitin Sulfate, Insulitis, Biomarkers
Abstract: Type 1 diabetes (T1D) is an autoimmune disease in which insulin-producing beta cells in pancreatic islets are progressively destroyed. Clinical trials of immunotherapies in recently diagnosed T1D patients have only transiently and partially impacted the disease course, suggesting that other approaches are required. Our previous studies have demonstrated that heparan sulfate (HS), a glycosaminoglycan conventionally expressed in extracellular matrix, is present at high levels inside normal mouse beta cells. Intracellular HS was shown to be critical for beta cell survival and protection from oxidative damage. T1D development in Non-Obese Diabetic (NOD) mice correlated with loss of islet HS and was prevented by inhibiting HS degradation by the endoglycosidase, heparanase. In this study we investigated the distribution of HS and heparan sulfate proteoglycan (HSPG) core proteins in normal human islets, a role for HS in human beta cell viability and the clinical relevance of intra-islet HS and HSPG levels, compared to insulin, in human T1D. In normal human islets, HS (identified by 10E4 mAb) co-localized with insulin but not glucagon and correlated with the HSPG core proteins for collagen type XVIII (Col18) and syndecan-1 (Sdc1). Insulin-positive islets of T1D pancreases showed significant loss of HS, Col18 and Sdc1 and heparanase was strongly expressed by islet-infiltrating leukocytes. Human beta cells cultured with HS mimetics showed significantly improved survival and protection against hydrogen peroxide-induced death, suggesting that loss of HS could contribute to beta cell death in T1D. We conclude that HS depletion in beta cells, possibly due to heparanase produced by insulitis leukocytes, may function as an important mechanism in the pathogenesis of human T1D. Our findings raise the possibility that intervention therapy with dual activity HS replacers/heparanase inhibitors could help to protect the residual beta cell mass in patients recently diagnosed with T1D.
Published: 2018

30. Non-invasive, in vitro analysis of islet insulin production enabled by an optical porous silicon biosensor

Author: Thomas W.H. Kay, Martin J. Sweetman, Rinku Chhasatia, Thomas Loudovaris, Nicolas H. Voelcker, Frances J. Harding, Michaela Waibel, Helen E. Thomas, Chhasatia, Rinku, Sweetman, Martin J, Harding, Frances J, Waibel, Michaela, Kay, Tom, Thomas, Helen, Loudovaris, Thomas, and Voelcker, Nicolas H
Subjects: Silicon, insulin, Aptamer, medicine.medical_treatment, Biomedical Engineering, Biophysics, Nanotechnology, 02 engineering and technology, Biosensing Techniques, macromolecular substances, 010402 general chemistry, Porous silicon, 01 natural sciences, Islets of Langerhans, Limit of Detection, PEG ratio, Electrochemistry, medicine, Humans, Insulin, Cells, Cultured, Detection limit, Chromatography, Base Sequence, optical biosensor, islet, Chemistry, Spectrum Analysis, technology, industry, and agriculture, Optical Devices, interferometric reflectance spectroscopy, aptamer, General Medicine, Aptamers, Nucleotide, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Transplantation, Glucose, Interferometry, porous silicon, Covalent bond, 0210 nano-technology, Biosensor, Antibodies, Immobilized, Porosity, Biotechnology
Abstract: A label-free porous silicon (pSi) based, optical biosensor, using both an antibody and aptamer bioreceptor motif has been developed for the detection of insulin. Two parallel biosensors were designed and optimised independently, based on each bioreceptor. Both bioreceptors were covalently attached to a thermally hydrosilylated pSi surface though amide coupling, with unreacted surface area rendered stable and low fouling by incorporation of PEG moieties. The insulin detection ability of each biosensor was determined using interferometric reflectance spectroscopy, using a range of different media both with and without serum. Sensing performance was compared in terms of response value, response time and limit of detection (LOD) for each platform. In order to demonstrate the capability of the best performing biosensor to detect insulin from real samples, an in vitro investigation with the aptamer-modified surface was performed. This biosensor was exposed to buffer conditioned by glucose-stimulated human islets, with the result showing a positive response and a high degree of selectivity towards insulin capture. The obtained results correlated well with the ELISA used in the clinic for assaying glucose-stimulated insulin release from donor islets. We anticipate that this type of sensor can be applied as a rapid point-of-use biosensor to assess the quality of donor islets in terms of their insulin production efficiency, prior to transplantation. Refereed/Peer-reviewed
Published: 2017

31. Linking Metabolic Abnormalities to Apoptotic Pathways in Beta Cells in Type 2 Diabetes

Author: Seth L. Masters, Helen E. Thomas, and Jibran A Wali
Subjects: Programmed cell death, endocrine system, Amyloid, endocrine system diseases, Bcl-2 pathway, 030209 endocrinology & metabolism, Review, Biology, medicine.disease_cause, 03 medical and health sciences, 0302 clinical medicine, medicine, oxidative stress, lcsh:QH301-705.5, 030304 developmental biology, 0303 health sciences, geography, geography.geographical_feature_category, Endoplasmic reticulum, Intrinsic apoptosis, apoptosis, Inflammasome, Type 2 diabetes, General Medicine, Islet, NLRP3 inflammasome, Cell biology, lcsh:Biology (General), Apoptosis, Immunology, pancreatic beta cell, endoplasmic reticulum stress, Oxidative stress, medicine.drug
Abstract: Pancreatic beta-cell apoptosis is an important feature of islets in type 2 diabetes. Apoptosis can occur through two major pathways, the extrinsic or death receptor mediated pathway, and the intrinsic or Bcl-2-regulated pathway. Hyperglycaemia, hyperlipidaemia and islet amyloid poly-peptide (IAPP) represent important possible causes of increased beta-cell apoptosis. Hyperglycaemia induces islet-cell apoptosis by the intrinsic pathway involving molecules of the Bcl-2 family. High concentrations of palmitate also activate intrinsic apoptosis in islets cells. IAPP oligomers can induce apoptosis by both intrinsic and extrinsic pathways. IL-1b produced through NLRP3 inflammasome activation can also induce islet cell death. Activation of the NLRP3 inflammasome may not be important for glucose or palmitate induced apoptosis in islets but may be important for IAPP mediated cell death. Endoplasmic reticulum (ER) and oxidative stress have been observed in beta cells in type 2 diabetes, and these could be the link between upstream metabolic abnormalities and downstream apoptotic machinery.
Published: 2013

32. Rotavirus acceleration of type 1 diabetes in non-obese diabetic mice depends on type I interferon signalling

Author: Barbara S. Coulson, Kate L. Graham, Helen E. Thomas, Fiona E. Fleming, Jessica A. Pane, and Thomas W.H. Kay
Subjects: Rotavirus, 0301 basic medicine, viruses, 030209 endocrinology & metabolism, Receptor, Interferon alpha-beta, Nod, Article, Rotavirus Infections, Diabetes Mellitus, Experimental, Mice, 03 medical and health sciences, 0302 clinical medicine, Immune system, Downregulation and upregulation, Mice, Inbred NOD, Interferon, medicine, Animals, NOD mice, Mice, Knockout, Type 1 diabetes, Multidisciplinary, business.industry, Pancreatic islets, virus diseases, Th1 Cells, medicine.disease, Virology, Diabetes Mellitus, Type 1, 030104 developmental biology, medicine.anatomical_structure, Interferon Type I, Immunology, business, Interferon type I, Signal Transduction, medicine.drug
Abstract: Rotavirus infection is associated with childhood progression to type 1 diabetes. Infection by monkey rotavirus RRV accelerates diabetes onset in non-obese diabetic (NOD) mice, which relates to regional lymph node infection and a T helper 1-specific immune response. When stimulated ex vivo with RRV, plasmacytoid dendritic cells (pDCs) from naïve NOD mice secrete type I interferon, which induces the activation of bystander lymphocytes, including islet-autoreactive T cells. This is our proposed mechanism for diabetes acceleration by rotaviruses. Here we demonstrate bystander lymphocyte activation in RRV-infected NOD mice, which showed pDC activation and strong upregulation of interferon-dependent gene expression, particularly within lymph nodes. The requirement for type I interferon signalling was analysed using NOD mice lacking a functional type I interferon receptor (NOD.IFNAR1−/− mice). Compared with NOD mice, NOD.IFNAR1−/− mice showed 8-fold higher RRV titers in lymph nodes and 3-fold higher titers of total RRV antibody in serum. However, RRV-infected NOD.IFNAR1−/− mice exhibited delayed pDC and lymphocyte activation, no T helper 1 bias in RRV-specific antibodies and unaltered diabetes onset when compared with uninfected controls. Thus, the type I interferon signalling induced by RRV infection is required for bystander lymphocyte activation and accelerated type 1 diabetes onset in genetically susceptible mice.
Published: 2016
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33. Complete Diabetes Protection Despite Delayed Thymic Tolerance in NOD8.3 TCR Transgenic Mice Due to Antigen-Induced Extrathymic Deletion of T Cells

Author: Grant Morahan, Jonathan Chee, Thomas W.H. Kay, Helen E. Thomas, David J. Izon, Pere Santamaria, Janette Allison, Kate L. Graham, Stacey Fynch, Gaurang Jhala, and Balasubramanian Krishnamurthy
Subjects: endocrine system, Endocrinology, Diabetes and Metabolism, Transgene, T-Lymphocytes, Antigen presentation, Receptors, Antigen, T-Cell, Mice, Transgenic, Thymus Gland, Biology, medicine.disease_cause, Lymphocyte Activation, Autoimmunity, 03 medical and health sciences, Mice, 0302 clinical medicine, Antigen, Mice, Inbred NOD, Internal Medicine, medicine, Cytotoxic T cell, Animals, 030304 developmental biology, NOD mice, 0303 health sciences, Peripheral Tolerance, Peripheral tolerance, Proteins, Diabetes Mellitus, Type 1, Immunology, Central Tolerance, Glucose-6-Phosphatase, Female, Central tolerance, Immunology and Transplantation, 030215 immunology
Abstract: Prevention of autoimmunity requires the elimination of self-reactive T cells during their development in the thymus and maturation in the periphery. Transgenic NOD mice that overexpress islet-specific glucose 6 phosphatase catalytic subunit–related protein (IGRP) in antigen-presenting cells (NOD-IGRP mice) have no IGRP-specific T cells. To study the relative contribution of central and peripheral tolerance mechanisms to deletion of antigen-specific T cells, we crossed NOD-IGRP mice to highly diabetogenic IGRP206–214 T-cell receptor transgenic mice (NOD8.3 mice) and studied the frequency and function of IGRP-specific T cells in the thymus and periphery. Peripheral tolerance was extremely efficient and completely protected NOD-IGRP/NOD8.3 mice from diabetes. Peripheral tolerance was characterized by activation of T cells in peripheral lymphoid tissue where IGRP was expressed followed by activation-induced cell death. Thymectomy showed that thymic output of IGRP-specific transgenic T cells compensated for peripheral deletion to maintain peripheral T-cell numbers. Central tolerance was undetectable until 10 weeks and complete by 15 weeks. These in vivo data indicate that peripheral tolerance alone can protect NOD8.3 mice from autoimmune diabetes and that profound changes in T-cell repertoire can follow subtle changes in thymic antigen presentation.
Published: 2012

34. A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy

Author: Helen E. Thomas, Helena Liang, Andrzej S. Januszewski, Alicia J. Jenkins, Alex W. Hewitt, Ryan J. Farr, Annie K. McAulley, Thomas Loudovaris, Anandwardhan A. Hardikar, Thomas W.H. Kay, and Mugdha V. Joglekar
Subjects: Multidisciplinary, Diabetic Retinopathy, business.industry, Gene Expression Profiling, Disease progression, High-Throughput Nucleotide Sequencing, Diabetic retinopathy, medicine.disease, Bioinformatics, DNA sequencing, Article, Gene expression profiling, Circulating MicroRNA, MicroRNAs, Mir-375, microRNA, Medicine, Biomarker (medicine), Cluster Analysis, Humans, Fluorescein Angiography, business, Biomarkers
Abstract: MicroRNAs are now increasingly recognized as biomarkers of disease progression. Several quantitative real-time PCR (qPCR) platforms have been developed to determine the relative levels of microRNAs in biological fluids. We systematically compared the detection of cellular and circulating microRNA using a standard 96-well platform, a high-content microfluidics platform and two ultra-high content platforms. We used extensive analytical tools to compute inter- and intra-run variability and concordance measured using fidelity scoring, coefficient of variation and cluster analysis. We carried out unprejudiced next generation sequencing to identify a microRNA signature for Diabetic Retinopathy (DR) and systematically assessed the validation of this signature on clinical samples using each of the above four qPCR platforms. The results indicate that sensitivity to measure low copy number microRNAs is inversely related to qPCR reaction volume and that the choice of platform for microRNA biomarker validation should be made based on the abundance of miRNAs of interest.
Published: 2015

35. The proapoptotic BH3-only proteins Bim and Puma are downstream of endoplasmic reticulum and mitochondrial oxidative stress in pancreatic islets in response to glucotoxicity

Author: Thomas W.H. Kay, Lut Overbergh, Dieter Rondas, Chantal Mathieu, Stacey Fynch, Jibran Ja Wali, Helen E. Thomas, Andreas Strasser, Mark D. McKenzie, Esteban Nicolas Gurzov, Yuxing Zhao, Shizuo Akira, and Lorraine Elkerbout
Subjects: 0301 basic medicine, Cancer Research, P53-UP-REGULATED MODULATOR, Ribose, Apoptosis, Mitochondrion, Endoplasmic Reticulum, Antioxidants, Tissue Culture Techniques, chemistry.chemical_compound, Mice, NEURONAL APOPTOSIS, Puma, Insulin-Secreting Cells, hemic and lymphatic diseases, oxidative stress, BETA-CELL APOPTOSIS, GENE-EXPRESSION, Mice, Knockout, biology, Bcl-2-Like Protein 11, Islets of Langerhans -- drug effects -- metabolism -- pathology, apoptosis, Oxidants -- pharmacology, ER STRESS, Sciences bio-médicales et agricoles, Endoplasmic Reticulum Stress, Oxidants, 3. Good health, Cell biology, Mitochondria, medicine.anatomical_structure, Original Article, Membrane Proteins -- deficiency -- genetics -- metabolism, type 2 diabetes, biological phenomena, cell phenomena, and immunity, ER stress, Life Sciences & Biomedicine, medicine.medical_specialty, Programmed cell death, Thapsigargin, Mice, 129 Strain, Immunology, Endoplasmic Reticulum -- drug effects -- metabolism -- pathology, INSULIN-SECRETION, Cell Line, Transcription Factor CHOP -- deficiency -- genetics, 03 medical and health sciences, Cellular and Molecular Neuroscience, Islets of Langerhans, HYDROGEN-PEROXIDE, Downregulation and upregulation, Oxidative Stress -- drug effects, Internal medicine, Proto-Oncogene Proteins, medicine, Animals, Humans, GLUCOSE TOXICITY, Bim, Ribose -- metabolism, RNA, Messenger, Science & Technology, Insulin-Secreting Cells -- metabolism -- pathology, Endoplasmic reticulum, Pancreatic islets, Tumor Suppressor Proteins, Endoplasmic Reticulum Stress -- drug effects, Membrane Proteins, DIABETES-MELLITUS, Cell Biology, Glucose -- metabolism, biology.organism_classification, RNA, Messenger -- metabolism, Tumor Suppressor Proteins -- deficiency -- genetics -- metabolism, Mice, Inbred C57BL, 030104 developmental biology, Endocrinology, Glucose, Antioxidants -- pharmacology, DNA-DAMAGE, chemistry, Diabetes Mellitus, Type 2, Diabetes Mellitus, Type 2 -- metabolism -- pathology, Mitochondria -- drug effects -- metabolism -- pathology, Unfolded protein response, Apoptosis Regulatory Proteins -- deficiency -- genetics -- metabolism, Proto-Oncogene Proteins -- deficiency -- genetics -- metabolism, Apoptosis Regulatory Proteins, Apoptosis -- drug effects, Transcription Factor CHOP
Abstract: Apoptosis of pancreatic beta cells is a feature of type 2 diabetes and its prevention may have therapeutic benefit. High glucose concentrations induce apoptosis of islet cells, and this requires the proapoptotic Bcl-2 homology domain 3 (BH3)-only proteins Bim and Puma. We studied the stress pathways induced by glucotoxicity in beta cells that result in apoptosis. High concentrations of glucose or ribose increased expression of the transcription factor CHOP (C/EBP homologous protein) but not endoplasmic reticulum (ER) chaperones, indicating activation of proapoptotic ER stress signaling. Inhibition of ER stress prevented ribose-induced upregulation of Chop and Puma mRNA, and partially protected islets from glucotoxicity. Loss of Bim or Puma partially protected islets from the canonical ER stressor thapsigargin. The antioxidant N-acetyl-cysteine also partially protected islets from glucotoxicity. Islets deficient in both Bim and Puma, but not Bim or Puma alone, were significantly protected from killing induced by the mitochondrial reactive oxygen species donor rotenone. Our data demonstrate that high concentrations of glucose induce ER and oxidative stress, which causes cell death mediated by Bim and Puma. We observed significantly higher Bim and Puma mRNA in islets of human donors with type 2 diabetes. This indicates that inhibition of Bim and Puma, or their inducers, may prevent beta-cell destruction in type 2 diabetes., info:eu-repo/semantics/published
Published: 2014

36. Hypoxia-Inducible Factor-1α (HIF-1α) Potentiates β-Cell Survival After Islet Transplantation of Human and Mouse Islets

Author: Thomas Loudovaris, Natasha A. Deters, Sue Mei Lau, Jenny E. Gunton, Kim Cheng, Rebecca A. Stokes, Thomas W.H. Kay, Frank J. Gonzalez, Philip J. O'Connell, Jessica Stolp, Wayne J. Hawthorne, Shane T. Grey, and Helen E. Thomas
Subjects: endocrine system, Transcription, Genetic, Cell Survival, Biomedical Engineering, Islets of Langerhans Transplantation, lcsh:Medicine, Apoptosis, Mice, SCID, Article, Diabetes Mellitus, Experimental, Mice, Diabetes mellitus, Medicine, Animals, Humans, Mice, Knockout, Transplantation, geography, geography.geographical_feature_category, business.industry, lcsh:R, Cell Biology, Hypoxia (medical), medicine.disease, Islet, Hypoxia-Inducible Factor 1, alpha Subunit, Cell Hypoxia, Deferoxamine, Mice, Inbred C57BL, surgical procedures, operative, Mechanism of action, Hypoxia-inducible factors, Cancer research, Female, medicine.symptom, business, medicine.drug
Abstract: A high proportion of β-cells die within days of islet transplantation. Reports suggest that induction of hypoxia-inducible factor-1α (HIF-1α) predicts adverse transplant outcomes. We hypothesized that this was a compensatory response and that HIF-1α protects β-cells during transplantation. Transplants were performed using human islets or murine β-cell-specific HIF-1α-null (β-HIF-1α-null) islets with or without treatment with deferoxamine (DFO) to increase HIF-1α. β-HIF-1α-null transplants had poor outcomes, demonstrating that lack of HIF-1α impaired transplant efficiency. Increasing HIF-1α improved outcomes for mouse and human islets. No effect was seen in β-HIF-1α-null islets. The mechanism was decreased apoptosis, resulting in increased β-cell mass posttransplantation. These findings show that HIF-1α is a protective factor and is required for successful islet transplant outcomes. Iron chelation with DFO markedly improved transplant success in a HIF-1α-dependent manner, thus demonstrating the mechanism of action. DFO, approved for human use, may have a therapeutic role in the setting of human islet transplantation.
Published: 2012

37. Granzyme B is dispensable in the development of diabetes in non-obese diabetic mice

Author: Joseph A. Trapani, Zia U.A. Mollah, Prerak Trivedi, Balasubramanian Krishnamurthy, Thomas W.H. Kay, Kate L. Graham, Thomas C. Brodnicki, and Helen E. Thomas
Subjects: Male, lcsh:Medicine, Autoimmunity, CD8-Positive T-Lymphocytes, Granzymes, Mice, Endocrinology, 0302 clinical medicine, Cell Movement, Mice, Inbred NOD, Molecular Cell Biology, Cytotoxic T cell, lcsh:Science, NOD mice, 0303 health sciences, Multidisciplinary, Cell Death, biology, T Cells, 3. Good health, medicine.anatomical_structure, Medicine, Female, Beta cell, Research Article, medicine.medical_specialty, Mice, 129 Strain, Immune Cells, Immunology, Thymus Gland, Autoimmune Diseases, Islets of Langerhans, 03 medical and health sciences, Internal medicine, medicine, Animals, Lymphocyte Count, Biology, Pancreas, Cell Proliferation, 030304 developmental biology, Diabetic Endocrinology, Pancreatic islets, lcsh:R, Diabetes Mellitus Type 1, medicine.disease, Mice, Inbred C57BL, Granzyme B, CTL, Diabetes Mellitus, Type 1, Granzyme, biology.protein, Clinical Immunology, lcsh:Q, Lymph Nodes, Insulitis, Spleen, T-Lymphocytes, Cytotoxic, 030215 immunology
Abstract: Pancreatic beta cell destruction in type 1 diabetes is mediated by cytotoxic CD8(+) T lymphoctyes (CTL). Granzyme B is an effector molecule used by CTL to kill target cells. We previously showed that granzyme B-deficient allogeneic CTL inefficiently killed pancreatic islets in vitro. We generated granzyme B-deficient non-obese diabetic (NOD) mice to test whether granzyme B is an important effector molecule in spontaneous type 1 diabetes. Granzyme B-deficient islet antigen-specific CD8(+) T cells had impaired homing into islets of young mice. Insulitis was reduced in granzyme B-deficient mice at 70 days of age (insulitis score 0.043±0.019 in granzyme B-deficient versus 0.139±0.034 in wild-type NOD mice p
Published: 2012

38. Autoreactive Cytotoxic T Lymphocytes Acquire Higher Expression of Cytotoxic Effector Markers in the Islets of NOD Mice after Priming in Pancreatic Lymph Nodes

Author: Thomas W.H. Kay, Balasubramanian Krishnamurthy, Kate L. Graham, Robyn Maree Slattery, Stacey Fynch, Helen E. Thomas, Pere Santamaria, and Zia U.A. Mollah
Subjects: Cytotoxicity, Immunologic, endocrine system, Antigen presentation, Priming (immunology), Biology, Major histocompatibility complex, Pathology and Forensic Medicine, Diabetes Mellitus, Experimental, Islets of Langerhans, Mice, Cross-Priming, Mice, Inbred NOD, Insulin-Secreting Cells, medicine, Cytotoxic T cell, Animals, NOD mice, Cell Proliferation, Inflammation, Antigen Presentation, Pancreatic islets, Regular Article, medicine.disease, Rats, Granzyme B, medicine.anatomical_structure, Immunology, Cancer research, biology.protein, Cytokines, Lymph Nodes, Insulitis, Biomarkers, T-Lymphocytes, Cytotoxic
Abstract: Cytotoxic T lymphocytes (CTLs) that cause type 1 diabetes are activated in draining lymph nodes and become concentrated as fully active CTLs in inflamed pancreatic islets. It is unclear whether CTL function is driven by signals received in the lymph node or also in the inflamed tissue. We studied whether the development of cytotoxicity requires further activation in islets. Autoreactive CTLs found in the islets of diabetes-prone NOD mice had acquired much higher expression of the cytotoxic effector markers granzyme B, interferon γ, and CD107a than had those in the pancreatic lymph node (PLN). Increased expression seemed to result from stimulation in the islet itself. T cells held up from migrating from the PLN by administration of the sphingosine-1-phosphate agonist FTY720 did not increase expression of cytotoxic molecules in the PLN. Stimulation did not require antigen presentation or cytokine secretion by the target β cells because it was not affected by the absence of class I major histocompatibility complex expression or by the overexpression of suppressor of cytokine signaling-1. Activation of CD40-expressing cells stimulated increased CTL function and β-cell destruction, suggesting that signals derived from CD40-expressing cells promote the acquisition of cytotoxicity in the islet environment. These data provide in vivo evidence that stimulation of cytotoxic effector molecule expression occurs in inflamed islets and is independent of β cells.
Published: 2011

39. 40(th) EASD Annual Meeting of the European Association for the Study of Diabetes : Munich, Germany, 5-9 September 2004

Author: S. Artigas, A V Dreval, Mark I. McCarthy, C Watson, Peter H. Bennett, M Quint, Y Ikeda, E Alpert, F Schiele, H Sekihara, Erik Gylfe, P Lowe, J Kuhlmann, Alain Golay, V Longo, Shahidul Alam Khan Akm., L G Mantovani, M Zawodniak-Szalapska, G Winkler, T Harrity, L Virág, U Johne, Kuo S-W., Linda C Tapsell, J Rodriguez, Michel Komajda, K Kankova, Carole A. Cull, M Sporna, E Estilles, U Ribel, M C Spruce, E Buzzigoli, T Prazak, J K McLaughlin, M K Lingohr, M Lim, F Calara, A Siebenhofer, G Meregalli, Roberto Anichini, A D Baron, R Kurashvili, P C Butler, G I Fantus, T. E. De Gooyer, Park Y-M., R. Walther, S Heinrich, Agnieszka Zawiejska, S Mukherjee, Nikolaos Papanas, G Wong, Ian D. Caterson, David M. Maahs, Shuichi Kaneko, Alexandra E. Butler, Francisco Javier Ampudia-Blasco, O N Kong, Attali J-R., C A Hedman, K Oshinyemi, Nicolle Müller, I C Cranston, N Okumus, M V Vlaiculescu, Balasubramanian Ravikumar, W W Cheatham, K Mukasa, K B Biswas, Annunziata Lapolla, Phil McEwan, G Mader, Gilles Chassot, Dragi Anevski, Werner A. Scherbaum, M Donath, C Hesselmann, R A Gandhi, David E. Moller, Ezio Bonifacio, C Garcia, V Ifandi, P Hornnes, Nieuwenhoven Fav., C Puech, S Pérez-Del-Pulgar, Kim S-R., G Hines, C Rubio Terrés, Michael Gaster, N. Hosszufalusi, A Scholze, Andrew A. Young, Stavros Liatis, F Hariri, S Tan, Paul Valensi, Allan E. Karlsen, J Kim, E. Moberg, J Kaiser, L Berman, G Nelson, A Altkrüger, P Kothare, D B Cook, S Doran, G. van Dijk, Shahnaz Shahinfar, Kim C-S., P Stahl, M Manousaki, S Sigrist, S K Lim, M. P. Stern, A Guberti, C Rezzani, J McKenney, Karl Thomaseth, Sofia Carlsson, M Julia, R Brillante, I Rubesova, T Darkow, E Matsumoto, Wendy M. Macfarlane, M Di Martino, G Bardini, Rossella Menghini, D Duhot, E Farcasiu, Annalisa Natalicchio, I Lindner, J Buvat, Christian L. Brand, Harry Dorchy, Iwona Pietrzak, Z T Luo, P Home, M Ekelund, Jesper Gromada, Kristine Færch, F Piarulli, H Kim, R Mentel, Zsuzsanna K. Zsengellér, Dullaart Rpf., Anton Luger, Thomas A. Pearson, V Manicardi, P Rösen, Feng Y-M., R Morganti, Lars Hansen, Demuth H-U., Haruo Kasai, A Shostak, Rudi Steffensen, G Taylor, Markolf Hanefeld, C Santini, E Hamaguchi, Roberto Miccoli, F Storms, M Cooper, Y Lee, Allison E. Aiello, P Smith, T Suehiro, K Treece, M Waluś, Timothy A Welborn, Simone Baltrusch, E Kontela, S Chai, J Crean, H Yokoyama, Johan G. Eriksson, Rafael Hernández Hernández, J Rodríguez-Saldaña, M P Tornero, G Formoso, D. Lovell, E Bingham, A Mylonakis, M Manteghetti, D Fedele, Antonio Martín-Duce, Ralph A. DeFronzo, D Salcedo, Kurt Højlund, Antonio Petrone, Sheu Whh., C Gutierrez, Flavia Pricci, S Kurita, Z G Abbas, M M Benedetti, Philippe A. Halban, Daniel J. Cox, O Ljungkvist, Justine Davies, J Palsgaard, Lars Sjöström, E Bosi, L Janin-Manificat, W. F. Kelly, M. Fernandez, E Colak, O V Mulyarchik, B Kronshage, F Lang, M Erfurth, Takashi Kadowaki, N Jendrike, U Walter, J Wishart, Y. Neye, D Kim, N Furuhashi, M Barsotti, D Florow, L Ke, L Borgquist, N C Jackson, Ffolliott M. Fisher, V Baskar, K Yoshioka, Bryan A. Wolf, G Chabrier, R Skoumal, Livio Luzi, H Kose, I Pharisien, B. Klein, H Winiarska, M C Johnson, L Griffiths, Nonna Kravchun, C Combe, Baptist Gallwitz, J Zdychova, L Skorda, Jorma Ilonen, W Gao, I N Steen, A Terrinoni, P D Ambery, W Kern, C M Kusminski, Cho M-H., Paolo Pozzilli, Louise G. Grunnet, E Schönle, David R Matthews, Robert W. Taylor, Y Cohen, Kim H-S., M P Eccles, N B Tutuncu, D McDowell, Richard M. Bergenstal, K Takamatsu, T Steiner, Jaan Palgi, Valdemar Grill, N Niculescu, G Federici, S Lehto, P. M. McKeigue, M Barone, Michael E. Trautmann, S Smirnov, J Mannion, M Eto, C Rousseau, M Conti, C S Ernest, Antonio Ceriello, D H Schweitzer, Jung E-D., Andreas Festa, Avijit Lahiri, A Shepelkevich, A Murro, A Kollmann, Jonathan R.S. Arch, R Landgraf, Son H-Y., I Engelsberger, E Agardh, S Rodríguez-Mulero, P J Kraml, K Lee, D. F. Du Toit, E Kim, G Fadini, Williams Ajk., Philip Home, M B Antcieferov, C Perlemoine, D Perrea, Song X-L., D Ruggieri, Krister Bokvist, Heidi Sørensen, Bilbao, G Yoshino, J P Taylor, Shen H-M., S M Furier, R Urquhart, J Wohlgelernter, Jianping Weng, T. Baba, Q Hong, C Silva, Castaigne J-P., M Felaco, X X Zhang, M Jaroň, Milla Rosengård-Bärlund, J G Papp, Toshio Miyata, Lervang H-H., Park M-K., I Kinalska, A Long, Oomen Phn., N Kogawa, Ippolita Patrizia Patera, S. Karadeniz, Dinesh Selvarajah, D S Chung, A Wensaas, Richard Imrich, M Recasens, J Ruxer, O Buchea, E Wilpart, S P Stepanenko, Le Ttd., H Ohgawara, Mariaconsuelo Valentini, A Mondok, M Peltonen, Marianne O. Larsen, K Chatzianagnostou, Agneta Ståhle, A L Ferrari, L Bordier, F Maingrette, A Matsuda, G Vukomanovic, Jakob D. Wikstrom, T Yamakita, E Gorostiaga, J Jin, B Gopalan, Heinz Drexel, S Hewitt, Rury R. Holman, C Dieterle, T L Ruchti, N Asatiani, M Sidira, A Iezzi, A J Sommerfield, D Châtenet, M L Olsen, R Bergemann, C Koehler, T L Kuraeva, B Balas, Christian Berne, E Santos-Mazo, G Smith, A Siejka, R Kožnarová, A Mattina, S Sheikh, A Adomeit, M Rasmussen, J. Fagerudd, N Busciantella Ricci, Nuria Vilarrasa, E Hammar, T L Thoms, L Aydın, Ron G. Rosenfeld, A Nikolajuk, R Gos, C L Morgan, H L Yu, D Dheelchand, S Ramrath, N Boudriga, Jerome I. Rotter, C Jahannault, W M Weston, Folke Lindgärde, M Hertlova, D Knight, A Monroy-Mayorga, E Pardini, A Chamson-Reig, B Franke, Janie McCluskey, Joseph Bryan, C Nikolopoulou, Christie M. Ballantyne, Fausto Santeusanio, L Pegoraro, M Lee, A Klimenko, S Jaiveer, K. Pettersson-Fernholm, Michael A. Nauck, A Ekbom-Schnell, G Deferrari, Riccardo Schiaffini, S. Pampanelli, Khan Aka., David Hopkins, Maija Wessman, M Kamarinos, Noh J-H., O Ebisui, K McCarroll, Jeppe Sturis, Peter Nowotny, N Gorbenko, Åke Sjöholm, David G. Maggs, A E Halseth, B Cresci, A A Ortiz-Gress, A Korakovouni, O Matejkova, C E Mogensen, C J Lin, Ramon Gomis, H Seaman, C Granier, Yang C-H., F Assah, O Sanchez, Fausto Machicao, Peter G. Morris, Alberto Ortiz, A Giardinelli, D Bracaglia, A Gonzalo, S Pavlatos, Andreas Lechner, F Canovic, L Sjolind, Allan Vaag, Birgitte Bruun Nielsen, David A. Ziegler, Vito Lampasona, R Gershoni-Baruch, A. Dei Cas, H Renz, E Mena, Matthew Waltham, Kim D-M., H Levanen, D D Mick, Valentina Alexandrovna Peterkova, E Meskhishvili, Sarah Nutland, R Bustani, John R. Lindsay, M Christoforidou, A Abicht, E Harno, K Cyganek, A Fitchet, S Neelotpol, P Nikishin, P Serradas, J Hinrichsen, M Halvorson, M Chovatia, B Voet, Jinny Willis, E Parretti, M Haslbeck, M Wellard, L Teng, Julio Wainstein, J S Fischer, K. Lalic, D Roggenland, I Gich, R Anwar, Maurizio Cassader, D Serota, X J Li, R J Schotzinger, Vilmundur Gudnason, Björn Zethelius, S A Wootton, W Andrzejewski, R Rezsohazy, R Gao, T Klimentova, T Mazurek, I Bruckner, C Dohrmann, R E James, G daSilva Xavier, Kim S-Y., A Dorca, Stuart J. Pocock, Terri J. Allen, I Giovos, P B Parab, N H Andersen, P Fotinakis, Miriam Cnop, H Lee, Norbert Tennagels, Omorodola I. Abatan, F Ailett, I. Lager, D Manzella, H Hut, Larry A. Distiller, G Lip, Lim S-K., Rong Zhang, T Tsuno, Steen Knudsen, M. Bajardi, Manuel Benito, Dai Sugimoto, Melvin J. Prince, D W Dunstan, D Rankins, K A Majali, G Ozansoy, Isabella Russo, S Uçak, G Annuzzi, R Talar-Wojnarowska, K Lange, S Neugebauer-Baba, Campbell H. Thompson, Eric Renard, P. D. Mountjoy, Z Morrison, Elizabeth A. Davis, Franco Cavallo, C Corvaja, R Antuña, Craig John Currie, H Linnebjerg, He Y-L., A J Palmer, Mariola R. Chacón, H Malinska, M. Jones, R Lichnovská, K Mandes, Paolo Tessari, T Mokhort, A Laina, H. L. Y. Chan, I Schmidt, R Banks, Richard G. IJzerman, L Ksinantova, G Setti, H Vaudry, A Gallo, V Spallone, Chen J-W., Thomas Danne, A Chong, M Hallschmid, S Aczel, S Hulme, N Islam, M Hosoi, P M Ternan, P Di Bartolo, N Bishara, T Shibasaki, Martin A. Osterhoff, Im S-S., M Jecht, T Hamaguchi, S Mattera, K Ways, Elizabeth Northam, U Rajala, Reinhard W. Holl, L Yang, S Panaiotopoulos, K Horvath, R Kluge, Thora B. Bodvarsdottir, Y Dong, Irene Alemanno, C McDougall, Reimar W. Thomsen, M Campbell, W Rabl, John Öhrvik, Yuichiro Yamada, Paola Ungaro, W Benzer, Mike Sampson, Roberto Trevisan, R G Radu, Aas A-M., P E Lobo, Ricardo Scott, S M Son, Josephine M. Forbes, T A Hillier, K L Wyne, Louis L. Nguyen, J Farmer, M H Tan, Kwon H-S., J Yang, L Sandvik, Franco Folli, A K Jenum, M Nguyen, W Pratipanawatr, A L Frederiksen, Rebecca Smith, Lee H-J., A Schäfer, C Manuelli, G S Denver, T Vukovich, B Maceira, K Matsumoto, K. Chokkalingam, Nurcan Üçeyler, P Modi, Timothy M. Morgan, S Mertens, B M Singh, Michaela Riedl, K Iso, C Cucurullo, G. F. Bottazzo, M Calvani, K Hur, J Wetzels, Kazuhiro Takahashi, Y Aso, H Stammer, M G Masding, Fitsum Guebre-Egziabher, J L González-Sánchez, L Armstrong, Alberto Maran, Peter G.F. Swift, S S Popovic, J Starczynski, E Vitacolonna, Luigi Laviola, R W Gelling, Marina Cardellini, D Barilla, Rosa de Diego Martínez, W H Landschulz, Anne Mette Rosenfalck, R K Wong, Kevin E. Schneider, K Peros, Giuseppe Nanni, F Zhang, I Rákóczi, T Iburi, M Nakhjavani, X Q Zhang, S Tournis, Per Lav Madsen, Graham A. Hitman, A. Tura, K Laubner, N D Kostic, Lawrence M. Dolan, R. Sinha Roy, J A Wagner, J. Tuomilehto, J Hauptman, M Abdel-Ghany, D Lacombe, Toralph Ruge, Johannes A Maassen, Triantafyllos Didangelos, K Sasaki, I Argüelles, Klaus Levin, C Popow, Emanuel Christ, R Chetty, L Baillet-Blanco, Jo-Ann Salmon, T Mine, James L. Trevaskis, I Franke, J Gorski, E A Andrianova, A Dayan, A Caballero, Aleksandra Gilis-Januszewska, M Yasujima, Z Kasalová, C.D.A. Stehouwer, F. K. Gorus, G A Nichols, A Glowania, David P. Strachan, P Fredlund, N. F. da Silva, P Reboldi, M Sausbier, K H Groenier, G Stuccio, N Guttman, K R Ahmed, A D Ristic, T Kapellen, J Coutcher, Aldo V. Greco, Oswald Wagner, A Zagayko, Maria Alevizaki, B B Zhang, W F Ferris, Jenny Fredriksson, Lois Jovanovic, J Hänninen, R De Giglio, Kazuo Yagui, O Potterat, P Hamliton, R E Scranton, B Mankovsky, A Stylianou, B Fellström, Abdel-Wahab Yha., M Kitagawa, Katherine L. Baldock, F R Johnson, F Baigts, S D'Addato, F J Sanz, A Mistry, S D Wise, T Pratipanawatr, U R Fölsch, James R.C. Parkinson, Claudia Sommer, C Park, F E Griffiths, M L Martí, R Demirtunc, S Taniguchi, J Lundkvist, T Siegmund, Juan Sztajzel, C Dienesch, F Baumgartner, L Scalone, T M Mckolanis, K Otake, Ullrik Pedersen-Bjergaard, T M Vriesendorp, Michael B. Wheeler, Henry Schmitt, Peter Hovind, S Lange, Stephane Roze, L. Van Gaal, B Klaproth, Anthony E. Civitarese, D Eckland, A Dagar, D F Hopkins, Kari Stefansson, C Gonzalez-Yanes, B Meyboom-de Jong, D. J. Betteridge, K Buhling, M Crepaldi, Ana M. Wägner, L Renna, L Volpe, R McBride, V Corbo, E O Brennesvik, R P Hayes, R Abdollahnia, G Viviani, C F Liew, Francisco Pérez-Bravo, Jeffrey Baron, Brian M. Frier, H H Samira, D Szentendrei, K. J. Schjoedt, W K Waldhäusl, D Gniuli, D Zou, G Tschank, V Urbančič, A L Nolan, Albertini J-P., J Malcomson, M Larbig, C Cheyssac, K Aurich, C M Kesson, S Heller, Maija E. Miettinen, R F Luco, Adrian J. Cameron, Luigi Mattiello, Z. Metelko, X E Zhang, M Parramón, I. G. 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Jensen, A D'Avanzo, J Monaghan, K P Yeo, Guivarch P-H., B Bauduceau, D Weghuber, P Tatti, J Ybarra, S Gwozdziewiczová, E Gasparini, B Saltin, Charlotte Granhall, Howard Leventhal, R Marin, M Tumiati, Cicero Afg., L Csémy, B Berger, S Mikros, D Dall'Asta, M Shahmanesh, Y G Vasiljev, F Potthoff, H S Randeva, G De Berardis, J O Logan, K Warncke, P Uitterlinden, E Rehring, K Gilmore, K Shankhdhar, V V Bojko, M Vahatalo, E A Korolyova, D Wiemann, P G Lankisch, D Hendrie, F Galtier, M Rybarczyk, Gisela Dahlquist, N N Rudovich, G Stein, A Liebl, F Tan, A Westerlund, S Gronemann, I Franklin, Jonathan A. Prince, Peter Arner, E Skliros, T. Sparre, M Vigas, Maddalena Trombetta, L. Bjerre Knudsen, A C Sima, I Dubroca, Alastair Gray, I Weets, R Ferraresi, Schauer Ujw., E. Leinonen, S Corazza, Jonathan Levy, P K Prakash, R Guzder, S. Barnhill, John Blangero, J Herreros, G. de Vries, Cheng Ptw., A Macías-Batista, K. 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Hansen, T Watanabe, Gang Hu, Malik Rja., T Kennedy-Martin, Ulla M Smidt, J.M. Dekker, A A Fisher, H Liu, R E Pratley, B Sun, C Fledelius, J F Raposo, R Langham, Ahn S-Y., B R Waterhouse, Shaoping Deng, W Ricart, S V Melnichenko, Henrike Sell, M. Tomalino, N Takeda, Paola Massucco, A Harlan, W Henrich, C D Byrne, R Junik, Khalid Hussain, D Pop Gorceva, Torben Hansen, C Ritterath, K Ogawa, D V Phan, Bradley S Metcalf, Robb E. Moses, Juan P. Frias, Hitoshi Ishii, C Brisard, P Wolkow, H H Maurer, Ingo Rustenbeck, Juris J. Meier, Octavian Savu, S D Lin, V B Bregovski, A Fox, S Cicala, M. Koenen, L Vignati, O de Divitiis, Constantin Ionescu-Tirgoviste, Kilian Rittig, J Song, Riccardo Candido, F Cohen-Boulakia, U Shankhdhar, P Jahan, Antonio Tiengo, E Liepinsh, C Álvarez, Sigurd Lenzen, James E. 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Chauveau, P, Combe, C, Baillet blanco, L, Beauvieux, M, Gin, H, Heinrich, S, Steiner, T, Ott, U, Holdass, H, Fellström, B, Jardine, A, Staffler, B, Logan, J, Gimpelewicz, C, Stanciu, C, Pena, C, Serafinceanu, C, González posada, J, Hernández, D, Pérez tamajó, L, Ló, A, Herná Alarcó, M, Meneses, M, Barsotti, M, Rizzo, G, Schmauss, S, Havrdova, T, Saudek, F, Boucek, P, Adamec, M, Invitti, C, Gilardini, L, Parati, G, Mazzilli, G, Pontiggia, B, Sartorio, A, Lutgers, H, Groenier, K, Zasadzinska, G, Saryusz wolska, M, Kurktschiev, D, Majdrakova, I, Varbanova, T, Todorova, B, Bajo martinez, A, Bernal, E, Sanchez, O, Ugalde canitrot, A, Sanchez largo, E, Coca robinot, D, Fabregate, R, Calbacho, M, Marquez, J, Saban ruiz, J, Penesova, A, Cizmarova, E, Blazicek, P, De Jongh, R, Serné, E, Ijzerman, R, De Vries, G, Andersen, N, Knudsen, S, Helleberg, K, Mogensen, C, Waluś, M, Idzior waluś, B, Sztefko, K, Cieślik, G, Fedak, D, Wozniakiewicz, E, Lin, S, Guo, M, Lin, C, Liu, X, Francisco, M, Rodriguez rosas, H, Peiró martinez, I, Macías batista, A, Harte, A, Rodriguez cuenca, S, Valsamakis, G, Chetty, R, Anderson, L, Roca, P, Matyka, K, Lasalle, J, Hershon, K, Berman, L, Gibson, E, Gillen, D, Maroni, J, Simmons, D, Hiukka, A, Forder, P, Leinonen, E, Hilden, H, Fruchart, J, Keech, A, Farnier, M, Freeman, M, Macdonell, G, Perevozskaya, I, Mitchel, Y, Gumbiner, B, Didangelos, T, Athyros, V, Mikhailidis, D, Papageorgiou, A, Bouloukos, V, Pehlivanidis, A, Symeonidis, A, Elisaf, M, Mckenney, J, Insull, W, Lewin, A, Maccubbin, D, Kush, D, Schuster, H, Barter, P, Stender, S, Bonnet, J, Morrell, J, Watkins, C, Kallend, D, Stalenhoef, A, Ballantyne, C, Murin, J, Tonstad, S, Rose, H, Wilpshaar, W, Jenkins, A, Karschimkus, C, Dragicevic, G, Rowley, K, Wolthers, T, Best, J, Voet, B, Murdoch, S, Marcovina, S, Brunzell, J, Caparevic, Z, Kostic, N, Ilic, S, Sartore, G, Piarulli, F, Cantaro, S, Reitano, R, Fiore, C, Marin, R, Bassan, S, Manzato, E, Fedele, D, Solini, A, Santini, E, Thornalley, P, Babaei jadidi, R, Karachalias, N, Kupich, C, Ahmed, N, Fowler, A, Baker, A, Starczynski, J, O'Hare, P, Szepietowska, B, Szelachowska, M, Puch, U, Glebocka, A, Quinn, D, Mcternan, C, Bonser, R, Idzior walus, B, Woźniakiewicz, E, Dimitriou, K, Apostolou, O, Kontela, E, Devangelio, E, Gould, E, Serri, O, Roussin, A, Buithieu, J, Mamputu, J, Renier, G, Giordanetti, S, De Amici, E, Poggi, G, Turpini, C, Fratino, P, Garzaniti, A, Banu, I, Paries, J, Roman, G, Negrean, M, Bala, C, Nita, C, Kistorp, C, Gustafsson, F, Chong, A, Lip, G, Galatius, S, Ari, N, Sahilli, M, Ceylan isık, A, Ozansoy, G, Karasu yilmaz, C, Matteucci, E, Rosada, J, Pallini, M, Evangelista, I, Cassetti, G, Giusti, C, Giampietro, O, Capaldo, B, Galderisi, M, Cicala, S, Turco, A, Imbroinise, A, Nosso, G, D'Errico, A, De Divitiis, O, Klimontov, V, Korolyova, E, Jeltova, L, Bondar, I, Tarkun, I, Arslan, B, Canturk, Z, Tarkun, P, Agacdiken, A, Komsuoglu, B, Méneveau, N, Pierre justin, E, Alsayed, M, Sabbah, R, Paulin, S, Marcu, S, Tauveron, I, Zimmermann, C, Schiele, F, Seronde, M, Vautrin, P, Lusson, J, Thieblot, P, Bernard, Y, Mistry, A, Pye, M, Peovska, I, Maksimovic Pavlovic, J, Vavlukis, M, Pop Gorceva, D, Bosevski, M, Scognamiglio, R, Negut, C, De Kreutzenberg, S, Madonna, R, De Caterina, R, Willerson, J, Geng, Y, Vahsen, S, Ledwig, D, Ramrath, S, Frantz, S, Schmidt, I, Calvillo, L, Dienesch, C, Elbing, I, Bischoff, H, Ertl, G, Bauersachs, J, Davydov, A, Mkrtum'Yan, A, Baranova, L, Ikeda, Y, Suehiro, T, Osaki, F, Ota, K, Arii, K, Kumon, Y, Hashimoto, K, Doney, A, Morris, A, Palmer, C, Byun, S, Doo, H, Pagnin, E, Calo, L, Fadini, G, Kubaszek, A, Chai, S, Chai, Q, Rasmussen, L, Ledet, T, Wogensen, L, Lengyel, C, Varró, A, Virág, L, Magyar, J, Bíró, T, Jost, N, Skoumal, R, Nánási, P, Tóth, M, Horkay, F, Papp, J, Zacharopoulou, O, Athanaselis, S, Tsokos, N, Doupis, J, Psallas, M, Cokkinos, D, Pavlatos, S, Liatis, S, Akhobadze, T, Dzneladze, L, Samarguliani, I, Taskiran, M, Rasmussen, V, Jensen, G, Fisher, A, Petrovsky, N, Srikusalanukul, W, Budge, M, Trifunovic zamaklar, D, Zivkovic, M, Jelic, V, Vukomanovic, G, Ristic, A, Seferovic, P, Costa, J, Duarte, S, Manley, S, Sailesh, S, Venkataraman, A, Haider, Y, Groza, I, Oprean, M, Ardelean, A, Morosanu, A, Darkow, T, Vanderplas, A, Mamas, M, Mcelduff, P, Burns, J, Edwards, R, Fitchet, A, Young, R, Gibson, J, Lichiardopol, R, Niculescu, N, Totora, A, Pencea, C, Tomescu, I, Cinteza, M, Manicardi, V, Coscelli, C, Navazio, A, Catellani, E, Michelini, M, Dall'Asta, D, Guberti, A, Piazza, A, Gasparini, E, Pantaleoni, M, Guiducci, U, Manari, A, Sejil, S, Janand delenne, B, Avierinos, J, Habib, G, Labastie, N, Vague, P, Lassmann vague, V, Luźniak, P, Tatoń, J, Wojciechowska Luźniak, A, Zairis, M, Lyras, A, Patsourakos, N, Tsirimbis, V, Foussas, S, Lupón, J, Urrutia, A, Herreros, J, González, B, Coll, R, Altimir, S, Prats, M, Valle, V, Abreu padí, C, Rábago, G, Ivanova, L, Brasacchio, D, Harno, E, Keenan, A, Li, H, Lu, Z, Ke, L, Liu, H, Jeong, I, Chae, M, Choi, M, Yoo, H, Kim, C, Yun, M, Na, M, Kang, Y, Kong, O, Son, S, Kim, I, Tanaka, N, Hosoi, M, Matsuyama, Y, Fukumoto, M, Yamakita, T, Yoshioka, K, Ishii, T, Sato, T, Fujii, S, Aoki, T, Shibata, T, Mizutani, N, Suzuki, J, Fowelin, J, Samuelsson, P, Brandrup wogsen, G, Okumura, K, Tokmakova, A, Staroverova, D, Antcieferov, M, Shutichina, I, Kuntchevich, G, Vriesendorp, T, Morélis, Q, Legemate, D, Schaper, F, Mainas, E, Gkioulmpasanis, I, Panagiotou, I, Vassilikos, G, Skorda, L, Sidira, M, Christoforidou, M, Alaveras, A, Artikis, V, Evdemon, E, Lechleitner, M, Koch, T, Ebenbichler, C, Sturm, W, Moretti, L, Moruzzo, D, Boldrini, E, Pandolfo, C, Kameyama, M, Iwasa, R, Cho, M, Nam, J, Huh, K, Kaplar, M, Paragh, G, Erdei, A, Csongradi, E, Garai, I, Varga, J, Galuska, L, Udvardy, M, Higa, M, Kaneko, Y, Hiroi, N, Koziarska, D, Nowacki, P, Majkowska, L, Luzniak, P, Wojciechowska luźniak, A, Tushuizen, M, Nieuwland, R, Snoeck, D, Sturk, A, Diamant, M, Aguiar, L, Bahia, L, Villela, N, Laflor, C, Conde, C, Bottino, D, Dorigo, D, Bouskela, E, Pu, S, Luo, Z, Lam, K, Dan, Q, Xu, A, Shen, J, Cheng, K, Xu, J, Thamer, C, Stefan, N, Haap, M, Heller, E, Tschritter, O, De Prado, A, Ortiz, A, Ybarra, J, Gich, I, Pou, J, Ehren, M, Roggenland, D, Reinsen, B, Klein, H, Rittig, K, Stock, J, Kocher, B, Balletshofer, B, Shon, H, Chung, D, Nakatani, Y, Matsuhisa, M, Kaneto, H, Hatazaki, M, Yoshiuchi, K, Katakami, N, Kawamori, D, Ohtoshi, K, Sakamoto, K, Matsuoka, T, Ozawa, K, Ogawa, S, Hori, M, Yamasaki, Y, Zitouni, K, Harry, D, Nourooz zadeh, J, Earle, K, Olesen, P, Franco, L, Corvaja, C, Semplicini, A, Ceylan işık, A, Arı, N, Rösen, P, Lee, I, Park, K, Jung, E, Shin, D, Jo, S, Obuobie, K, Prakash, P, Hanna, F, Lazarus, J, Varadhan, L, Gurushankar, J, James, D, Sheikh, S, Gaede, P, Zou, D, Vilarrasa, N, Perez maraver, M, Mena, E, Perez, D, Setti, G, Buckingham, R, Urbančič, V, Stefanovska, A, Bernjak, A, Ažman juvan, K, Kocijančič, A, Glowania, A, Filters, T, Fosmark, D, Torjesen, P, Kilhovd, B, Berg, T, Sandvik, L, Hanssen, K, Mentink, C, Donchenko, G, Stepanenko, S, Maingrette, F, Deng, H, Lindenmair, A, Freudenthaler, A, Baumgartner parzer, S, Nizheradze, K, Khoruzhenko, A, Tronko, N, Sheu, W, Ou, H, Shen, H, Lin, T, Wu, H, Yang, C, Mogylnytska, L, Schmoelzer, I, Davies, J, Band, M, Struthers, A, Prázný, M, Škrha, J, Kasalová, Z, Neelotpol, S, Jahan, P, Kauschke, S, Harrop, C, Schäfer, A, Widder, J, Eigenthaler, M, Walter, U, Uchimura, I, Ikebukuro, M, Kaibara, M, Hirata, M, Helal, R, Pervin, F, Yang, X, Jansson, P, Nagaev, I, Jack, M, Carvalho, E, Sunnerhagen, K, Cam, M, Cushman, S, Smith, U, Creely, S, Farmer, J, Gustafson, B, Kusminski, C, Krusinova, E, Wohl, P, Klementova, M, Lanska, V, Mcdougall, C, Kelly, I, Abbas, Z, Lutale, J, Archibald, L, Karunajeewa, H, Stingemore, N, Stuccio, G, Mcgechie, D, Muller, L, Hak, E, Goudzwaard, W, Montorsi, F, Homering, M, Sprenger, K, Goldstein, I, Asnaghi, V, Ferrari, G, Rastaldi, M, Gabellini, D, Dell'Antonio, G, Maestroni, A, Ruggieri, D, Luzi, L, Piemonti, L, Zerbini, G, Anafaroglu, I, Tutuncu, N, Sultana, M, Siddiqua, N, Iwasaki, T, Nakajima, A, Yoneda, M, Mukasa, K, Tanaka, S, and Sekihara, H
Subjects: 0303 health sciences, medicine.medical_specialty, business.industry, EASD, Endocrinology, Diabetes and Metabolism, Human physiology, medicine.disease, 03 medical and health sciences, 0302 clinical medicine, Diabetes mellitus, Family medicine, Internal Medicine, Medicine, business, 030217 neurology & neurosurgery, 030304 developmental biology
Published: 2004

40. Perforin and Fas induced by IFNγ and TNFα mediate beta cell death by OT-I CTL.

Author: Mark D McKenzie, Nadine L Dudek, Lina Mariana, Mark MW Chong, Joseph A Trapani, Thomas WH Kay, and Helen E Thomas
Abstract: Direct interaction between auto-reactive CTL and specific peptide–MHC class I complexes on pancreatic beta cells is critical in mediating beta cell destruction in type I diabetes. We used mice with genetic modifications in three major pathways used by CTL, perforin, Fas and pro-inflammatory cytokines to assess the relative contribution of these mechanisms to beta cell death. In vitro-activated ovalbumin (OVA)-specific CTL, from OT-I TCR-transgenic mice, specifically killed transgenic beta cells expressing OVA (from RIP-mOVA mice) in a 16-h cytotoxicity assay. Perforin-deficient CTL had a reduced ability to kill OVA-expressing islets in vitro (22.1 ± 3.8%) compared with wild-type CTL (71.4 ± 4.6%). Fas-deficient islets were only slightly protected from wild-type CTL but were completely protected from the residual killing observed with perforin-deficient CTL. Residual cytotoxicity in perforin-deficient CTL was also prevented by overexpression of SOCS-1, which blocks multiple cytokine signaling pathways. It was also prevented by pre-incubation with anti-tumor necrosis factor-alpha (anti-TNFα) antibody or by blocking IFNγ responsiveness through expressing a dominant negative IFNγ receptor. Perforin-deficient CTL produced IFNγ and TNFα that was shown to directly induce islet Fas expression during the assays. This suggests that Fas-deficiency, SOCS-1 overexpression and blockade of IFNγ and TNFα all protect beta cells from residual cytotoxicity of perforin-deficient CTL by blocking Fas upregulation. These findings indicate that wild-type CTL destroy antigen-expressing islets via a perforin-dependent mechanism. However, in the absence of perforin, the Fas/FasL pathway provides an alternative mechanism dependent on islet cell Fas upregulation by cytokines IFNγ and TNFα. [ABSTRACT FROM AUTHOR]
Published: 2006
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41. Diabetes induced by checkpoint inhibition in nonobese diabetic mice can be prevented or reversed by a JAK1/JAK2 inhibitor

Author: Tingting Ge, Amber‐Lee Phung, Gaurang Jhala, Prerak Trivedi, Nicola Principe, David J De George, Evan G Pappas, Sara Litwak, Laura Sanz‐Villanueva, Tara Catterall, Stacey Fynch, Louis Boon, Thomas W Kay, Jonathan Chee, Balasubramanian Krishnamurthy, and Helen E Thomas
Subjects: cytokines, CD8+ T cells, immune checkpoint inhibitor‐induced diabetes, JAK inhibitor, NOD mice, Immunologic diseases. Allergy, RC581-607
Abstract: Abstract Objectives Immune checkpoint inhibitors have achieved clinical success in cancer treatment, but this treatment causes immune‐related adverse events, including type 1 diabetes (T1D). Our aim was to test whether a JAK1/JAK2 inhibitor, effective at treating spontaneous autoimmune diabetes in nonobese diabetic (NOD) mice, can prevent diabetes secondary to PD‐L1 blockade. Methods Anti‐PD‐L1 antibody was injected into NOD mice to induce diabetes, and JAK1/JAK2 inhibitor LN3103801 was administered by oral gavage to prevent diabetes. Flow cytometry was used to study T cells and beta cells. Mesothelioma cells were inoculated into BALB/c mice to induce a transplantable tumour model. Results Anti‐PD‐L1‐induced diabetes was associated with increased immune cell infiltration in the islets and upregulated MHC class I on islet cells. Anti‐PD‐L1 administration significantly increased islet T cell proliferation and islet‐specific CD8+ T cell numbers in peripheral lymphoid organs. JAK1/JAK2 inhibitor treatment blocked IFNγ‐mediated MHC class I upregulation on beta cells and T cell proliferation mediated by cytokines that use the common γ chain receptor. As a result, anti‐PD‐L1‐induced diabetes was prevented by JAK1/JAK2 inhibitor administered before or after checkpoint inhibitor therapy. Diabetes was also reversed when the JAK1/JAK2 inhibitor was administered after the onset of anti‐PD‐L1‐induced hyperglycaemia. Furthermore, JAK1/JAK2 inhibitor intervention after checkpoint inhibitors did not reverse or abrogate the antitumour effects in a transplantable tumour model. Conclusion A JAK1/JAK2 inhibitor can prevent and reverse anti‐PD‐L1‐induced diabetes by blocking IFNγ and γc cytokine activities. Our study provides preclinical validation of JAK1/JAK2 inhibitor use in checkpoint inhibitor‐induced diabetes.
Published: 2022
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42. Development of a standardized MRI protocol for pancreas assessment in humans.

Author: John Virostko, Richard C Craddock, Jonathan M Williams, Taylor M Triolo, Melissa A Hilmes, Hakmook Kang, Liping Du, Jordan J Wright, Mara Kinney, Jeffrey H Maki, Milica Medved, Michaela Waibel, Thomas W H Kay, Helen E Thomas, Siri Atma W Greeley, Andrea K Steck, Daniel J Moore, and Alvin C Powers
Subjects: Medicine, Science
Abstract: Magnetic resonance imaging (MRI) has detected changes in pancreas volume and other characteristics in type 1 and type 2 diabetes. However, differences in MRI technology and approaches across locations currently limit the incorporation of pancreas imaging into multisite trials. The purpose of this study was to develop a standardized MRI protocol for pancreas imaging and to define the reproducibility of these measurements. Calibrated phantoms with known MRI properties were imaged at five sites with differing MRI hardware and software to develop a harmonized MRI imaging protocol. Subsequently, five healthy volunteers underwent MRI at four sites using the harmonized protocol to assess pancreas size, shape, apparent diffusion coefficient (ADC), longitudinal relaxation time (T1), magnetization transfer ratio (MTR), and pancreas and hepatic fat fraction. Following harmonization, pancreas size, surface area to volume ratio, diffusion, and longitudinal relaxation time were reproducible, with coefficients of variation less than 10%. In contrast, non-standardized image processing led to greater variation in MRI measurements. By using a standardized MRI image acquisition and processing protocol, quantitative MRI of the pancreas performed at multiple locations can be incorporated into clinical trials comparing pancreas imaging measures and metabolic state in individuals with type 1 or type 2 diabetes.
Published: 2021
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43. Replacing murine insulin 1 with human insulin protects NOD mice from diabetes.

Author: Colleen M Elso, Nicholas A Scott, Lina Mariana, Emma I Masterman, Andrew P R Sutherland, Helen E Thomas, and Stuart I Mannering
Subjects: Medicine, Science
Abstract: Type 1, or autoimmune, diabetes is caused by the T-cell mediated destruction of the insulin-producing pancreatic beta cells. Non-obese diabetic (NOD) mice spontaneously develop autoimmune diabetes akin to human type 1 diabetes. For this reason, the NOD mouse has been the preeminent murine model for human type 1 diabetes research for several decades. However, humanized mouse models are highly sought after because they offer both the experimental tractability of a mouse model and the clinical relevance of human-based research. Autoimmune T-cell responses against insulin, and its precursor proinsulin, play central roles in the autoimmune responses against pancreatic beta cells in both humans and NOD mice. As a first step towards developing a murine model of the human autoimmune response against pancreatic beta cells we set out to replace the murine insulin 1 gene (Ins1) with the human insulin gene (Ins) using CRISPR/Cas9. Here we describe a NOD mouse strain that expresses human insulin in place of murine insulin 1, referred to as HuPI. HuPI mice express human insulin, and C-peptide, in their serum and pancreata and have normal glucose tolerance. Compared with wild type NOD mice, the incidence of diabetes is much lower in HuPI mice. Only 15-20% of HuPI mice developed diabetes after 300 days, compared to more than 60% of unmodified NOD mice. Immune-cell infiltration into the pancreatic islets of HuPI mice was not detectable at 100 days but was clearly evident by 300 days. This work highlights the feasibility of using CRISPR/Cas9 to create mouse models of human diseases that express proteins pivotal to the human disease. Furthermore, it reveals that even subtle changes in proinsulin protect NOD mice from diabetes.
Published: 2019
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44. Loss of intra-islet heparan sulfate is a highly sensitive marker of type 1 diabetes progression in humans.

Author: Charmaine J Simeonovic, Sarah K Popp, Lora M Starrs, Debra J Brown, Andrew F Ziolkowski, Barbara Ludwig, Stefan R Bornstein, J Dennis Wilson, Alberto Pugliese, Thomas W H Kay, Helen E Thomas, Thomas Loudovaris, Fui Jiun Choong, Craig Freeman, and Christopher R Parish
Subjects: Medicine, Science
Abstract: Type 1 diabetes (T1D) is an autoimmune disease in which insulin-producing beta cells in pancreatic islets are progressively destroyed. Clinical trials of immunotherapies in recently diagnosed T1D patients have only transiently and partially impacted the disease course, suggesting that other approaches are required. Our previous studies have demonstrated that heparan sulfate (HS), a glycosaminoglycan conventionally expressed in extracellular matrix, is present at high levels inside normal mouse beta cells. Intracellular HS was shown to be critical for beta cell survival and protection from oxidative damage. T1D development in Non-Obese Diabetic (NOD) mice correlated with loss of islet HS and was prevented by inhibiting HS degradation by the endoglycosidase, heparanase. In this study we investigated the distribution of HS and heparan sulfate proteoglycan (HSPG) core proteins in normal human islets, a role for HS in human beta cell viability and the clinical relevance of intra-islet HS and HSPG levels, compared to insulin, in human T1D. In normal human islets, HS (identified by 10E4 mAb) co-localized with insulin but not glucagon and correlated with the HSPG core proteins for collagen type XVIII (Col18) and syndecan-1 (Sdc1). Insulin-positive islets of T1D pancreases showed significant loss of HS, Col18 and Sdc1 and heparanase was strongly expressed by islet-infiltrating leukocytes. Human beta cells cultured with HS mimetics showed significantly improved survival and protection against hydrogen peroxide-induced death, suggesting that loss of HS could contribute to beta cell death in T1D. We conclude that HS depletion in beta cells, possibly due to heparanase produced by insulitis leukocytes, may function as an important mechanism in the pathogenesis of human T1D. Our findings raise the possibility that intervention therapy with dual activity HS replacers/heparanase inhibitors could help to protect the residual beta cell mass in patients recently diagnosed with T1D.
Published: 2018
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45. Activation of the NLRP3 inflammasome complex is not required for stress-induced death of pancreatic islets.

Author: Jibran A Wali, Esteban N Gurzov, Stacey Fynch, Lorraine Elkerbout, Thomas W Kay, Seth L Masters, and Helen E Thomas
Subjects: Medicine, Science
Abstract: Loss of pancreatic beta cells is a feature of type-2 diabetes. High glucose concentrations induce endoplasmic reticulum (ER) and oxidative stress-mediated apoptosis of islet cells in vitro. ER stress, oxidative stress and high glucose concentrations may also activate the NLRP3 inflammasome leading to interleukin (IL)-1β production and caspase-1 dependent pyroptosis. However, whether IL-1β or intrinsic NLRP3 inflammasome activation contributes to beta cell death is controversial. This possibility was examined in mouse islets. Exposure of islets lacking functional NLRP3 or caspase-1 to H2O2, rotenone or thapsigargin induced similar cell death as in wild-type islets. This suggests that oxidative or ER stress do not cause inflammasome-mediated cell death. Similarly, deficiency of NLRP3 inflammasome components did not provide any protection from glucose, ribose or gluco-lipotoxicity. Finally, genetic activation of NLRP3 specifically in beta cells did not increase IL-1β production or cell death, even in response to glucolipotoxicity. Overall, our results show that glucose-, ER stress- or oxidative stress-induced cell death in islet cells is not dependent on intrinsic activation of the NLRP3 inflammasome.
Published: 2014
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46. Granzyme B is dispensable in the development of diabetes in non-obese diabetic mice.

Author: Zia U Mollah, Kate L Graham, Balasubramanian Krishnamurthy, Prerak Trivedi, Thomas C Brodnicki, Joseph A Trapani, Thomas W Kay, and Helen E Thomas
Subjects: Medicine, Science
Abstract: Pancreatic beta cell destruction in type 1 diabetes is mediated by cytotoxic CD8(+) T lymphoctyes (CTL). Granzyme B is an effector molecule used by CTL to kill target cells. We previously showed that granzyme B-deficient allogeneic CTL inefficiently killed pancreatic islets in vitro. We generated granzyme B-deficient non-obese diabetic (NOD) mice to test whether granzyme B is an important effector molecule in spontaneous type 1 diabetes. Granzyme B-deficient islet antigen-specific CD8(+) T cells had impaired homing into islets of young mice. Insulitis was reduced in granzyme B-deficient mice at 70 days of age (insulitis score 0.043±0.019 in granzyme B-deficient versus 0.139±0.034 in wild-type NOD mice p
Published: 2012
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