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

• 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
• 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

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（T1 D）是一种自身免疫性疾病，在美国困扰着近200万人。 2在胰腺中产生β细胞导致绝对需要注射胰岛素，造成重大风险 三是死亡率和发病率。T1 D的特征在于潜伏（无症状）期，在此期间，自身免疫或 4炎症性胰腺β细胞损伤被假定导致β细胞功能/质量的下降，并最终 五是T1 D。T1 D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。的 6种精确的胰腺内信号传导机制，导致免疫系统和早期胰腺炎的激活 7人受伤情况仍不清楚。鉴定与这些关键免疫事件密切相关的标记物， 8可以在外周循环中检测到的T1 D的发病机制将允许检测临床前 9疾病和疾病轨迹的跟踪。细胞外囊泡（EV）及其内容物已经成为新的 人类代谢疾病中细胞间信号传导和功能性生物标志物的10种介质。信息的途径 11个合作小组作为NIH EV生物学工作的一部分（NIH共同基金）表明，循环细胞特异性 12电动汽车及其货物作为疾病轨迹或细胞健康的探针，比传统的 全血浆中的13种循环RNA或蛋白质生物标志物。最近的T1 D研究表明，胰腺β细胞 14在“压力”下产生含有自身抗原和RNA转录物的EV， 15之间的胰腺和免疫细胞的分子货物的转移。然而，研究表明， T1 D中胰腺β细胞衍生EV的16种功能景观及其作为T1 D生物标志物的意义 17儿童期易感性，缺乏。针对RFA-DK-21-016，我们假设胰岛细胞- 18种衍生的EV是胰岛细胞生物学的功能性报告基因，并且含有与免疫调节相关的RNA货物。 T1 D早期的19种反应和β细胞健康。在目标1中，我们利用成熟的人类细胞系统， 20例胰腺损伤（人细胞系和捐献的人胰岛，伴和不伴精氨酸介导的损伤） 21与我们小组建立的方法一起提供β- 22个细胞来源的EV，从患有和 23没有T1 D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。 24项功能（巨噬细胞、中性粒细胞）通过评估被假定为胰腺损伤的早期介质 25免疫代谢表型和对体内施用人EV至糖尿病易感者的反应 26模型系统。在目标3中，我们将使用RNA-seq和生物信息学来鉴定胰腺β细胞特异性 来自长期NIDDK TEDDY研究（n=140，1：1）的儿童的27个与T1 D事件相关的转录本 28例T1 D病例/对照），T1 D免疫风险高。完成后，此应用程序将提供广泛的 人类β细胞衍生EV的29种表型、功能和临床表征，以开发一种特征 30例与早期T1 D发展相关的胰腺细胞特异性EV，解决了RFA-DK-21-016的目的。