Project Abstracts - CBDS

Martha Campbell-Thompson, DVM, PhD, Investigator, University of Florida
Clayton Mathews, PhD, co-Investigator, University of Florida
Mark Atkinson, PhD, co-Investigator, University of Florida
Hao Chen, PhD, co-Investigator, University of Tennessee

Start Date: September 19, 2014

End Date:  August 31, 2018

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Abstract

Our studies are focused on examination of single islet gene expression profiles (transcriptomes) to determine pathways critical to human beta cell death and survival.  In preliminary studies we have developed new approaches and protocols that allow laser-capture of individual islets from cryosections of human pancreas, extract the RNA, and obtain comprehensive transcriptomes.  Our proposal to the HIRN-CBDS consortium is based on using and expanding these proven protocols to gain deeper insights into islet heterogeneity and differences in gene expression patterns of individual islets from organ donors with and without diabetes as well as with or without serum autoantibodies (AAB). Based on preliminary data in islets from non-diabetic individuals with or without AAB, we hypothesize that islets develop a gene expression signature of “cellular stress” before the early stages of insulitis. A full characterization of early abnormalities in islet gene expression signatures could point to novel mechanisms of disease and new therapeutic strategies that target islet abnormalities to delay or prevent beta cell death. The use of comprehensive gene expression profiling of thousands of expressed genes will allow a very detailed look at specific pathways and processes. Our proposal has the following specific aims: Specific Aim 1 is to characterize transcriptomes of individual islets in a survey of islets from different regions of the pancreas and from organ donors with or without autoantibodies (AAB) and with or without diabetes. This will provide an unbiased survey of islet gene expression and its heterogeneity. In Specific Aim 2 we will characterize transcriptomes of individual islets selected by immunolocalization for insulin and CD3+ cells. Additionally, in Specific Aim 3 we seek to characterize transcriptomes of individual islets selected for staining patterns that mark the presence of viruses and/or metabolic stress.  In this aim we will use our newly acquired ability to conduct multiplexed immunofluorescence to define islets with novel complex phenotypes. In this aim phenotypes will be highly complex and we will stain one tissue section and then capture individual islets of interest from adjacent serial tissue sections. Mechanistic studies using isolated islets will follow from all specific aims as new pathways of cellular destruction or survival are discovered.

Publications

• Increased Inflammation as well as Decreased Endoplasmic Reticulum Stress and Translation Differentiate Pancreatic Islets of Pre-symptomatic Stage 1 Type 1 Diabetes and Non-diabetic Cases
• Alpha cell receptor for advanced glycation end products associate with glucagon expression in type 1 diabetes
• Adaptation to chronic ER stress enforces pancreatic β-cell plasticity
• The IGFBP3/TMEM219 pathway regulates beta cell homeostasis
• What is a β cell? – Chapter I in the Human Islet Research Network (HIRN) Review Series
• Genetic predisposition in the 2′-5’A pathway in the development of type 1 diabetes: potential contribution to dysregulation of innate antiviral immunity
• Islet sympathetic innervation and islet neuropathology in patients with type 1 diabetes
• NKG2D Signaling Within the Pancreatic Islets Reduces NOD Diabetes and Increases Protective Central Memory CD8(+) T Cell Numbers
• What the HLA-I!-Classical and Non-classical HLA Class I and Their Potential Roles in Type 1 Diabetes
• Type 1 diabetes alters ischemia-induced gene expression
• The Role of Accessory Cells in Islet Homeostasis
• Islet Microvasculature Alterations With Loss of Beta-cells in Patients With Type 1 Diabetes
• Abnormal islet sphingolipid metabolism in type 1 diabetes
• Pancreatic Histopathology of Human Monogenic Diabetes Due to Causal Variants in KCNJ11, HNF1A, GATA6, and LMNA
• Neuropeptide FF increases M2 activation and self-renewal of adipose tissue macrophages
• Insulitis in Autoantibody-Positive Pancreatic Donor With History of Gestational Diabetes Mellitus
• Islet-Derived CD4 T Cells Targeting Proinsulin in Human Autoimmune Diabetes
• Factors That Influence the Quality of RNA From the Pancreas of Organ Donors
• Re-addressing the 2013 consensus guidelines for the diagnosis of insulitis in human type 1 diabetes: is change necessary?
• Analysis of self-antigen specificity of islet-infiltrating T cells from human donors with type 1 diabetes
• Islet cell hyperexpression of HLA class I antigens: a defining feature in type 1 diabetes
• Insulitis and β-Cell Mass in the Natural History of Type 1 Diabetes
• Organ donor specimens: What can they tell us about type 1 diabetes?
• Nebivolol: a multifaceted antioxidant and cardioprotectant in hypertensive heart disease