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Co-registration of Cell Organization, Phenotype and Function in the Human Pancreas During Type 1 Diabetes

1 型糖尿病期间人类胰腺细胞组织、表型和功能的共同注册

基本信息

批准号：
10490416
负责人：
MARK A. ATKINSON
金额：
$ 50.68万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-17 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10490416
关键词：
3-Dimensional Acinar Cell Address Age Agonist Alpha Cell Amylases Antibodies Area Arginine Autoantibodies Autoimmunity Autophagocytosis Beta Cell Biological Assay Blood Vessels Carbachol Caring Cell Death Cell physiology Cells Cellular Assay Cellular Stress Characteristics Cholecystokinin Chromatin Computer Analysis Cytometry Data Defect Detection Deterioration Development Diabetes Mellitus Diabetes autoantibodies Disease Disease Progression Endocrine Enhancers Environment Enzymes Epigenetic Process Epitopes Exocrine pancreas Extravasation Failure Functional disorder GLP-I receptor Glucagon Glucose Histology Hormones Human Image Imatinib Immune Impairment In Situ Individual Infiltration Inflammation Inflammatory Insulin Insulin-Dependent Diabetes Mellitus Intervention Islet Cell Islets of Langerhans Laboratories Link Lipase Location Lymphatic Measures Messenger RNA Metabolic Pathway Methodology Molecular Molecular Profiling Natural History Oligonucleotides Organ Organ Donor Pancreas Pathology Pathway interactions Persons Phenotype Physiological Principal Investigator Production Proinsulin Proteins Proteomics Recovery Residencies Resolution Role Series Serum Signal Pathway Signal Transduction Slice Stimulus Structure of beta Cell of islet System Techniques Testing Therapeutic Intervention Time Tissue-Specific Gene Expression Tissues Transplantation Transposase Trypsin Trypsinogen Tumor-infiltrating immune cells Tyrosine Kinase Inhibitor antibody conjugate base cell type cellular imaging cytokine diabetes pathogenesis diabetogenic endoplasmic reticulum stress epigenomics exenatide experience indexing innovation insight insulitis islet islet cell antibody live cell imaging non-diabetic novel prevent programs reconstruction response senescence single-cell RNA sequencing targeted imaging therapeutic candidate therapeutic target tool trafficking transcriptome transcriptomics

项目摘要

Recent evidence put forth by our group and others suggests type 1 diabetes (T1D) pathogenesis involves a combination of immune, islet, and acinar pancreas defects. In addition to autoimmunity and β-cell death, it has become clear that T1D is characterized by a whole-organ pathology with reduced pancreas size, reduced exocrine enzyme levels in serum, and altered α- and β-cell function, including impaired insulin processing, even in the islet autoantibody positive (AAb+) pre-T1D condition. Hence, there is a need to understand each of these facets in concert, linking cellular phenotype and function, together with studies of the human pancreas tissue microenvironment, throughout T1D progression. We hypothesize that alterations to β-cell status and its surrounding environment are key determinants of impaired β-cell function, exocrine function, and infiltration (insulitis). We propose to assay islet and acinar tissue function using our novel pancreas slice culture platform (Aim 1a) to test (pro)hormone (proinsulin, insulin, glucagon) and enzyme (lipase, trypsinogen) secretion from T1D, AAb+, and control organ donor pancreata in response to established endocrine (glucose, arginine, KCl) and exocrine stimuli (cholecystokinin, carbachol). We will correlate these functional data with molecular features via scRNAseq (single cell RNA sequencing) with antibody-based CITEseq (Cellular Indexing of Transcriptomes and Epitopes) and scATACseq (single cell assay for transposase-accessible chromatin sequencing); this, for the purpose of cell identification together with transcriptomic and epigenomic analyses (Aims 1b). Pancreas slices will also be subjected to these same stimulatory conditions for live cellular imaging of Ca2+ signalling activity within islet and acinar tissue areas in real time (Aim 2a), then fixed and analyzed by imaging mass cytometry (IMC). We will assess in situ expression of 120 immune and pancreas cell markers with cellular resolution (Aim 2b), followed by spatial and temporal analysis of IMC data to determine how islet, immune and acinar cell phenotypes correlate with tissue and cellular function, using our histoCAT (histology topography cytometry analysis toolbox). This will enable computational analysis with 3D reconstruction from serial sections (Aim 2c). Finally, in Aim 3, human donor pancreas slices will be subjected to diabetogenic stimuli (inflammatory cytokines, glucotoxicity) and interventions targeting β-cell stress [imatinib (tyrosine kinase inhibitor), MSL-7 (autophagy enhancer), exenatide (GLP-1 receptor agonist)], and similarly evaluated by single cell and IMC profiling. With over 14 years of experience in procurement of transplant-quality human pancreata through the Network for Pancreatic Organ donors with Diabetes (nPOD), we are uniquely poised to perform the proposed studies. We expect to identify altered molecular pathways and tissue features linking β-cell, whole-islet, and acinar cell phenotypes with cellular function in AAb+ and T1D pancreata. We anticipate these same defects will arise in slices subjected to diabetogenic stimuli, providing a platform to test known and novel candidates for targeted intervention to reduce β-cell stress, restore islet and acinar cell function, as well as prevent disease progression.

最近我们的研究小组和其他人提出的证据表明，1型糖尿病（T1D）的发病机制涉及a