Stage-specific Beta Cell Response and Biomarker Profile During Virus-induced T1D

病毒诱导的 T1D 期间阶段特异性 β 细胞反应和生物标志物概况

• 批准号: 9176017
• 负责人: DALE Leslie GREINER
• 金额: $ 37.69万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9176017
• 关键词: Address; Animal Model; Autoimmune Process; Autoimmunity; Beta Cell; Biological Markers; Biopsy; Blood Vessels; Cell Death; Cells; Cellular Stress; Cessation of life; Data; Development; Diabetes Mellitus; Disease; Early Diagnosis; Event; Foundations; Future; Gene Expression; Heterogeneity; Human; Hyperglycemia; Immune; Immunodeficient Mouse; Individual; Infection; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Islet Cell; Kinetics; Laboratories; Lead; MHC Class I Genes; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Profiling; Organ; Oxidative Stress; Pancreas; Pathogenesis; Pathology; Pathway interactions; Phase; Plasma; Prevention strategy; Process; Prognostic Marker; Publishing; Rattus; Report (document); Small RNA; Specimen; Staining method; Stains; Stress; Structure of beta Cell of islet; Technology; Testing; Time; Virus; Virus Diseases; Whole Blood; Work; base; circulating biomarkers; circulating microRNA; diabetic rat; diabetogenic; experimental study; immune activation; islet; mRNA Expression; microRNA biomarkers; new technology; novel strategies; novel therapeutic intervention; potential biomarker; public health relevance; response; response biomarker; single cell sequencing; transcriptome; transcriptome sequencing

 DESCRIPTION (provided by applicant): The mechanism(s) underlying T1D initiation and progression remain elusive as individuals are asymptomatic and difficult to identify and study during the early stages of disease. Even if biomarkers could identify humans early in the disease process, pancreas specimens are not readily available to permit direct study of the events that occur in the target organ. To address this, we propose to use the virus-inducible BBDR rat model of T1D. The BBDR rat is the only natural animal model that develops T1D following viral infection. Its close similarity with human T1D pathology, availability of new approaches for studying beta cell stress, and novel technologies for RNAseq analyses of purified beta cells and for identifying microRNA (miRNA) signatures in plasma now permit systematic identification of events underlying early stages of T1D. Diabetes in BBDR rats occurs in four stages: 1) initiation of beta cell stress and 2) death during the pre-insulitis period. Inslitis (autoimmunity, stage 3), and T1D (stage 4) occurs after complete beta cell destruction. We hypothesize that beta cell stress and death occurs prior to the initiation of insulitis and that plasma biomarkers reflecting beta cell stress will be detectable during this "pre-insulitis" stage f disease. Aim 1 will identify mRNA and microRNA signatures in beta cells at discrete stages of T1D development. We hypothesize that mRNA and microRNA beta cell expression profiles reflect the molecular mechanisms that are operational during each stage of progression of T1D. We will use RNAseq to generate mRNA and microRNA profiles to identify the molecular pathways that underlie beta cell responses at defined stages of T1D. Aim 2 will interrogate islet cell heterogeneity using single cell sequencing technology and identify mRNA profiles of beta cells with disparate MHC class I or insulin expression. We hypothesize that beta cell responses to stress during the development of T1D are heterogeneous, and that this may lead to the "spotty" insulitis and beta cell loss observed in rat (and human) T1D. We will use single cell RNAseq analyses of isolated islet cells to interrogate gene expression changes that underlie beta cell heterogeneity at defined stages of diabetes progression and correlate mRNA signatures with MHC class I or insulin expression. Aim 3 will identify stage-specific plasma microRNA signatures as T1D biomarkers. We hypothesize that the miRNA profile in the plasma will provide stage-specific signatures as biomarkers of T1D progression. We will use novel technology to recover small RNA from circulating plasma and perform RNAseq in the virus-induced BBDR rat and in our new model of Coxsackie B4-infected immunodeficient mice engrafted with human islets that will permit us to identify human miRNA signatures. These studies will permit detailed analyses of the mechanisms underlying beta cell decay prior to the development of insulitis and diabetes and associate them with ongoing events in the beta cells, leading to the identification of fundamental factors that underlie the earliest events of the disease. This will lay the foundation for future studies to look for miRNA biomarker signatures in humans that may be predisposed to T1D.

 描述（由适用提供）：由于个体在疾病的早期阶段不对称且难以识别和研究，因此T1D启动和进展的机制仍然难以捉摸。即使生物标志物可以在疾病过程的早期识别人类，胰腺标本也不容易可用，可以直接研究目标器官中发生的事件。为了解决这个问题，我们建议使用病毒诱导的BBDR大鼠BBDR大鼠是唯一在病毒感染后发展T1D的天然动物模型。它与人类T1D病理学，研究β细胞应激的新方法的可用性以及用于纯化β细胞的RNASEQ分析的新技术以及等离子体中的microRNA（miRNA）特征的新技术，现在可以系统地识别T1D早期阶段的事件的系统识别。 BBDR大鼠中的糖尿病分为四个阶段：1）β细胞应激的倡议和2）在胰岛炎前期的死亡。插管炎（自身免疫，第3阶段）和T1D（第4阶段）发生在完全β细胞破坏后。我们假设β细胞应激和死亡发生在胰岛素的主动性之前发生，并且在这种“胰岛素前炎” F期F疾病期间，可以检测到反映β细胞应激的血浆生物标志物。 AIM 1将在T1D开发的离散阶段鉴定β细胞中的mRNA和microRNA特征。我们假设mRNA和microRNAβ细胞表达谱图反映了在T1D进展阶段在每个阶段运行的分子机制。我们将使用RNASEQ生成mRNA和microRNA曲线，以确定在T1D定义阶段的β细胞反应构成的分子途径。 AIM 2将使用单细胞测序技术询问胰岛细胞的异质性，并鉴定具有不同MHC I类或胰岛素表达的β细胞的mRNA谱。我们假设在T1D发育过程中，β细胞对应激的反应是异质的，这可能导致在大鼠（和人）T1D中观察到的“斑点”胰岛素胰岛素和β细胞损失。我们将使用分离的胰岛细胞的单细胞RNASEQ分析来询问基因表达变化，这些变化是β细胞的异质性基础的基础糖尿病进展阶段，并将mRNA特征与MHC I类或胰岛素表达相关的mRNA特征。 AIM 3将识别特异性等离子体microRNA特征为T1D生物标志物。我们假设等离子体中的miRNA谱将提供特定于阶段的特异性特征，以作为T1D进展的生物标志物。我们将使用新型技术从循环血浆中恢复小RNA，并在病毒诱导的BBDR大鼠中进行RNASEQ，并在我们的新的Coxsackie B4感染的免疫缺陷小鼠的新模型中，与人类胰岛互动，使我们能够识别人类miRNA签名。这些研究将允许在胰岛细胞和糖尿病发展之前详细分析β细胞衰减的机制，并将其与β细胞中持续的事件相关联，从而鉴定出疾病最早事件的基本因素。这将为将来的研究奠定基础，以寻找可能易于T1D的人类中的miRNA生物标志物特征。