Project Abstracts - CBDS

Lori Sussel, PhD, Investigator, University of Colorado, Denver
Kristin Burnum-Johnson,  PhD, co-Investigator, Pacific Northwest National Laboratories
Julia Laskin, PhD, co-Investigator, Purdue University
Thomas Metz, PhD, co-Investigator, Pacific Northwest National Laboratory
Galya Orr, PhD, co-Investigator, Pacific Northwest National Laboratory

Start Date: September 22, 2015
End Date: May 31, 2020

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Abstract

Type 1 diabetes mellitus (T1D) is a chronic disease resulting from the autoimmune destruction of insulin-producing pancreatic beta cells. In addition to evolving ideas about the role of beta cells in genetic susceptibility to T1D, there is mounting evidence of diverse beta cell responses to T-cell mediated immune attack. In particular, several recent studies in mice, with some corresponding results in humans, suggest that beta cell stress and dysfunction may not only lead to beta cell death, but could possibly lead to increased beta cell plasticity and/or dedifferentiation. Understanding when and how these responses occur will not only identify potential new biomarkers of T1D, but will facilitate the development of novel therapies to prevent or block disease progression. For this reason, there is growing impetus to characterize the altered molecular and metabolic signature of diabetes-susceptible beta cells at early stages in the disease process. To facilitate the characterization of the cellular and molecular events that are responsible for beta cell dysfunction and destruction early in the T1D process there is a need to develop novel omics technologies that can be used to evaluate available human tissues. Furthermore, given the heterogeneity associated with normal and genetically susceptible T1D islets, it will be critical to develop analytical tools that allow detailed characterization of human pancreatic tissues at single cell and near single cell resolution. In accordance with these goals, the purpose of this grant is to use two powerful and complementary technology platforms – combFISH and nanoDESI – for in situ single cell resolution omics analyses of human pancreatic tissue (that includes gene and protein expression, metabolite and lipid abundances), supplemented and informed with laser capture microdissection-coupled proteomics and transcriptomics to provide critical insights into T1D. Our approach is to first optimize existing omics techniques on pancreatic islets derived from rodent models of T1D while the main thrust of the program will be to evaluate human normal and T1D islets and document and disseminate the data to the diabetes community. We will accomplish our goal through two primary aims: (1) Technology Refinement in Rodent Models of diabetes and (2) Application of Technology to Human Islet Studies.

Publications

• Regulation of β-cell death by ADP-ribosylhydrolase ARH3 via lipid signaling in insulitis
• Single Molecule-based FliFISH Validates Radial and Heterogeneous Gene Expression Patterns in Pancreatic Islet β Cells
• Pancreatic β cell regeneration: To β or not to β
• Comprehensive Proteomics Analysis of Stressed Human Islets Identifies GDF15 as a Target for Type 1 Diabetes Intervention
• The Long Noncoding RNA Paupar Modulates PAX6 Regulatory Activities to Promote Alpha Cell Development and Function
• High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labelling in a Nanodroplet Sample Preparation Platform
• FISHing for β Cells
• New mass spectrometry technologies contributing towards comprehensive and high throughput omics analyses of single cells
• Proteomic Analysis of Single Mammalian Cells Enabled by Microfluidic Nanodroplet Sample Preparation and Ultrasensitive NanoLC-MS
• High Spatial Resolution Imaging of Mouse Pancreatic Islets Using Nanospray Desorption Electrospray Ionization Mass Spectrometry
• Fluctuation Localization Imaging-based Fluorescence in situ Hybridization (fliFISH) for Accurate Detection and Counting of RNA Copies in Single Cells
• Heterogeneity of the Pancreatic Beta Cell