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：
10517890
负责人：
Saumya Das
金额：
$ 82万
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/10517890
关键词：
Address Autoantigens Autocrine Communication Autoimmune Autoimmune Diseases Autoimmunity B-Lymphocytes Beta Cell Bioinformatics Biological Markers Biological Models Biology Blood Blood Circulation Bone Marrow Case-Control Studies Cell Line Cell Nucleus Cell physiology Cells Cellular biology Characteristics Child Childhood Clinical Communication Communities Data Data Set Detection Development Diabetes Mellitus Discrimination Disease 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 Lead Measures Mediating Mediator of activation protein 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 Play 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 base biomarker identification case control 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 macrophage monocyte mortality neutrophil novel peripheral blood pre-clinical prevent protein biomarkers response 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 1 型糖尿病 (T1D) 是一种自身免疫性疾病，困扰着美国近 200 万人。 2 胰腺中产生β细胞导致绝对需要注射胰岛素，从而造成重大风险 3 T1D 的死亡率和发病率的特点是潜伏（无症状）阶段，在此期间存在自身免疫或免疫性疾病。 4 炎症性胰腺 β 细胞损伤被认为会导致 β 细胞功能/质量下降，并最终导致 5 T1D 的一个关键目标是通过限制免疫介导的损伤来阻止自身免疫细胞攻击。 6 种精确的胰腺内信号传导机制，可激活免疫系统和早期胰腺 7 损伤与这些关键免疫事件密切相关的标记物的鉴定仍不清楚。 8 可以在外周循环中检测到的 T1D 发病机制将允许临床前检测 9 疾病和疾病轨迹追踪已成为新奇事物。人类代谢疾病中的 10 种细胞间信号传导介质和功能生物标志物。 11 个合作小组作为 NIH 在 EV 生物学方面的工作（NIH 共同基金）的一部分表明，循环细胞特异性 12 EV 及其货物作为疾病轨迹或细胞健康的探针提供比传统方法更高的特异性全血浆中的 13 种循环 RNA 或蛋白质生物标志物最近对 T1D 的研究表明胰腺 β 细胞。 14 在“压力”下产生含有自身抗原和可能介导通讯的RNA转录本的EV 15 然而，研究描述了胰腺细胞和免疫细胞之间通过分子货物转移的情况。 16 型 T1D 中胰腺 β 细胞衍生的 EV 的功能状况及其作为 T1D 生物标志物的意义 17 儿童时期的易感性缺乏根据 RFA-DK-21-016，我们与胰岛细胞作斗争。 18 个衍生的 EV 具有功能性产生胰岛细胞生物学功能，并含有与免疫调节相关的 RNA 货物 19 T1D 早期反应和 β 细胞健康在目标 1 中，我们利用成熟的人类细胞系统。 20 胰腺损伤（人类细胞系和捐赠的人类胰岛，有或没有细胞因子介导的损伤） 21 以及我们小组建立的方法来提供 β- 的蛋白质组学和转录表征 22 细胞衍生的 EV，从患有以下疾病的儿童的人体循环中分离出胰腺 β 细胞特异性 EV 23 没有 T1D 在目标 2 中，我们定义了胰腺 β 细胞特异性 EV 在先天免疫中的功能作用。通过评估推测 24 种功能（巨噬细胞、中性粒细胞）可作为胰腺损伤的早期介质 25 种免疫代谢表型以及对易患糖尿病的体内给予人 EV 的反应 26 模型系统在目标 3 中，我们将使用 RNA-seq 和生物信息学来识别胰腺 β 细胞特异性。来自长期 NIDDK TEDDY 研究的儿童的 27 份与 T1D 事件相关的转录本（n=140，1:1 28 T1D 病例/对照）具有 T1D 高免疫风险。完成后，该应用将提供广泛的支持。人 β 细胞衍生的 EV 的 29 个表型、功能和临床特征，旨在开发特征 30 种与早期 T1D 发展相关的胰腺细胞特异性 EV，解决了 RFA-DK-21-016 的目标。