Characterization of beta-cell-specific extracellular vesicle cargo as functional biomarkers for type I DM disease

β细胞特异性细胞外囊泡货物作为 I 型 DM 疾病功能性生物标志物的表征

基本信息

批准号：
10706576
负责人：
Saumya Das
金额：
$ 75.37万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-19 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10706576
关键词：
Address Autoantigens Autocrine Communication Autoimmune Autoimmune Diseases Autoimmunity B-Lymphocytes Beta Cell Bioinformatics Biological Markers Biological Models Biology Blood Bone Marrow Case/Control Studies Cell Line Cell physiology Cells Cellular biology Characteristics Child Childhood Circulation Clinical Communication Communities Data Data Set Detection Development Diabetes Mellitus Discrimination Disease Early Diagnosis Event Funding Genetic Transcription Goals Health Histology Human Human Cell Line Immune Immune response Immune system Immunologics In Vitro Individual Inflammation Inflammatory Injury Insulin Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Laboratories Macrophage Measures Mediating Mediator Membrane Membrane Potentials Membrane Proteins Metabolic Diseases Metabolism Methods Mitochondria Modeling Molecular Morbidity - disease rate Mus National Institute of Diabetes and Digestive and Kidney Diseases Natural Immunity Oxidative Stress Pancreas Pancreatic Injury Pathogenesis Pathologic Patients Peripheral Persons Phase Phenotype Plasma Population Predisposition Process Production Proteins Proteomics RNA RNA Databases Reporter Research Risk Role Sentinel Signal Transduction Specificity Stimulus Stress Structure of beta Cell of islet Synapses System Techniques Tissues Transcript Translations United States National Institutes of Health cell injury cohort cytokine diabetes pathogenesis disease diagnostic early detection biomarkers extracellular vesicles glucose uptake human tissue immune activation immunoregulation in vivo insight insulin dependent diabetes mellitus onset intercellular communication islet lipid metabolism monocyte mortality neutrophil novel peripheral blood pre-clinical prevent protein biomarkers response single nucleus RNA-sequencing trafficking transcriptome sequencing translational study

项目摘要

1 Type 1 diabetes (T1D) is an autoimmune disease afflicting nearly 2 million people in the U.S. The loss of insulin- 2 producing β cells in the pancreas results in an absolute requirement for injected insulin, causing significant risks 3 of mortality and morbidity. T1D is characterized by a latent (asymptomatic) phase, during which autoimmune or 4 inflammatory pancreatic beta cell injury is postulated to lead to a decline in beta cell function/mass and ultimately 5 to T1D. A key goal in T1D is halting the autoimmune cellular attack, by limiting immune-mediated damage. The 6 precise intrapancreatic signaling mechanisms that lead to activation of the immune system and early pancreatic 7 injury remain unclear. Identification of markers closely associated with these key immune events in the 8 pathogenesis of T1D that can be detected in peripheral circulation would allow for detection of pre-clinical 9 disease and tracking of disease trajectory. Extracellular vesicles (EVs) and their contents have emerged as novel 10 mediators of intercellular signaling and functional biomarkers in human metabolic diseases. Data from our 11 collaborative group as part of NIH efforts in EV biology (NIH Common Fund) suggest that circulating cell-specific 12 EVs and their cargo as probes for disease trajectory or cellular health provide greater specificity than traditional 13 circulating RNA or protein biomarkers in whole plasma. Recent studies in T1D suggest that pancreatic beta cells 14 under “stress” produce EVs containing auto-antigens and RNA transcripts that may mediate communication 15 between pancreatic and immune cells by transfer of molecular cargo. Nevertheless, studies characterizing the 16 functional landscape of pancreatic beta-cell-derived EVs in T1D, and their implications as biomarkers of T1D 17 susceptibility in childhood, are lacking. In response to RFA-DK-21-016, we hypothesize that pancreatic islet cell- 18 derived EVs are functional reporters of islet cell biology and contain RNA cargo relevant to regulation of immune 19 responses and beta cell health early in T1D. In Aim 1, we utilize well-established human cellular systems of 20 pancreatic injury (human cell-line and donated human islets, with and without cytokine-mediated injury) 21 alongside methods established by our group to provide proteomic and transcriptional characterization of beta- 22 cell-derived EVs, with isolation of pancreatic beta-cell specific EVs from human circulation in children with and 23 without T1D. In Aim 2, we define the functional role for pancreatic beta-cell specific EVs in innate immune 24 function (macrophages, neutrophils) postulated to serve as early mediators of pancreatic injury via assessments 25 of immunometabolic phenotypes and responses to in vivo administration of human EVs to a diabetes-prone 26 model system. In Aim 3, we will use RNA-seq and bioinformatics to identify pancreatic beta cell-specific 27 transcripts associated with incident T1D from children from the long-standing NIDDK TEDDY study (n=140, 1:1 28 T1D case/control) at high immunologic risk for T1D. Upon completion, this application will provide a broad 29 phenotypic, functional and clinical characterization of human beta cell derived EVs toward developing a signature 30 of pancreatic cell-specific EVs relevant to early T1D development, addressing the aims of RFA-DK-21-016.

1型糖尿病（T1D）是一种自身免疫性疾病，在美国遭受近200万人 2在胰腺中产生β细胞会导致对注射胰岛素的绝对需求，从而引起很大的风险 3的死亡率和发病率。 T1D的特征是潜在的（无症状）阶段，在此期间自动免疫或 4炎症性胰腺β细胞损伤被张贴导致β细胞功能/质量的下降，最终导致 5至T1D。 T1D的关键目标是通过限制免疫介导的损伤来停止自身免疫性细胞攻击。这 6个精确的综合信号传导机制，导致免疫系统激活和早期胰腺激活 7伤害仍不清楚。与这些关键免疫事件紧密相关的标记物的识别 8可以在周围循环中检测到的T1D发病机理将允许检测到临床前 9疾病和疾病轨迹的追踪。细胞外蔬菜（EV）及其内容已成为新颖人类代谢疾病中的10个细胞间信号传导和功能生物标志物的介体。来自我们的数据 11合作小组作为NIH EV生物学努力（NIH Common Fund）的一部分，表明循环细胞特异性 12 ev及其作为疾病轨迹或细胞健康问题的货物比传统提供了更大的特异性 13整个血浆中循环RNA或蛋白质生物标志物。 T1D的最新研究表明胰腺β细胞 14在“应力”下产生的电动汽车，其中包含自动抗原和RNA转录本，可能介导通信 15通过分子货物的转移在胰腺和免疫细胞之间。然而，描述的研究 16 T1D中胰腺β细胞衍生的电动汽车的功能景观，它们作为T1D的生物标志物的含义缺乏17个童年的敏感性。响应RFA-DK-21-016，我们假设胰岛细胞 - 18派生的电动汽车是胰岛细胞生物学的功能记者，并包含与免疫调节有关的RNA货物 T1D早期的19个反应和β细胞健康。在AIM 1中，我们利用了公认的人类细胞系统 20胰腺损伤（人类细胞线和捐赠的人类胰岛，有和没有细胞因子介导的损伤） 21以及我们小组建立的方法提供β-提供蛋白质组学和转录表征 22个细胞衍生的电动汽车，通过与患有和 23没有T1D。在AIM 2中，我们定义了先天免疫中胰腺β细胞特异性电动汽车的功能作用 24个功能（巨噬细胞，嗜中性粒细胞）通过评估作为胰腺损伤的早期介体 25个免疫代谢表型和对人体内电动汽车对糖尿病易经的反应 26型号系统。在AIM 3中，我们将使用RNA-seq和生物信息学识别胰腺β细胞特异性长期存在的NIDDK Teddy研究的儿童的27种与事件T1D相关的成绩单（n = 140，1：1 28 T1D病例/对照）T1D具有高免疫学风险。完成后，本申请将提供广泛的 29人类β细胞衍生的EV的表型，功能和临床表征，以开发一个特征胰腺细胞特异性电动汽车的30与早期T1D发育有关，以解决RFA-DK-21-016的目的。