Dissection of cellular interactions with integrated functional genomics in comple

项目总结/摘要 认识到系统方法对复杂疾病的作用，在最初的资助期间， 开发了高度可控的三色阵列和分析平台，专注于采集质量数据。 我们的目标是将这些工具应用于导致自身免疫的事件，使用的是一种独特的衍生物， 1型糖尿病（T1DM）的生物育种（BB）大鼠模型。BB DRlyp/lyp大鼠是自发性糖尿病 由于Gimap5基因突变，使它们缺乏调节性T（Treg）细胞。DR +/+大鼠 具有完整的Gimap 5基因，不会发生自发性T1DM，但在消耗后会变成糖尿病 Treg细胞BB大鼠中的T1DM还涉及细胞毒性T细胞，因为它们的耗竭具有保护作用。我们的基因 糖尿病前期胰腺淋巴结（PLN）的表达研究表明，肥大细胞终末 在DRlyp/lyp动物中激活与在DR +/+动物中负调控;我们已经证实了它们的 通过两种不同的肥大细胞抑制药物预防大鼠T1DM的功能作用。因此我们 假设除了细胞毒性T效应细胞外，肥大细胞在DRlyp/lyp中起关键早期作用 糖尿病发生，尽管有疾病倾向，但Treg细胞足以预防DR +/+中的T1DM 大鼠我们进一步假设与这些状态相关的是不同的外周细胞因子环境，因为我们 发现来自T1DM风险人群的血清或在发病时收集的血清诱导特征性基因 健康外周血单核细胞（PBMC）中不同于正常人的表达特征 对照组和长期存在的T1DM。重要的是，我们发现观察这种分子特征长达5年 在T1DM发作之前。BB大鼠的可预测的疾病过程使得能够纵向解剖 免疫细胞亚群的活动，并检查这些活动是如何表现在周边。因此 我们建议：1）在时间和空间上剖析抗原呈递、调节和效应的活动 通过表达谱分析和组织学方法在规定的时间点观察细胞。2)WF、DR +/+血清 和DRlyp/lyp大鼠将纵向评估诱导健康PBMC中基因表达的因素。 将确认候选介质和疾病时间点特异性生物标志物的存在。理解 肥大细胞、T效应细胞和Treg细胞在自身免疫中的关系， 为免疫调节建立新的范例，并为人类疾病创造新的见解。

Project Summary/Abstract Recognizing the power of a systems approach to complex disease, during the initial funding period we developed a highly-controlled three color array and analysis platform focused on acquisition of quality data. Our goal is apply these tools to the events leading to autoimmunity using a unique derivative of the BioBreeding (BB) rat model of type 1 diabetes mellitus (T1DM). BB DRlyp/lyp rats are spontaneously diabetic due to a mutation in the Gimap5 gene, rendering them deficient in regulatory T (TREG) cells. DR+/+ rats possess an intact Gimap5 gene and do not develop spontaneous T1DM, but become diabetic upon depletion of TREG cells. T1DM in BB rats also involves cytotoxic T cells, since their depletion is protective. Our gene expression studies of prediabetic pancreatic lymph nodes (PLN) have shown that mast cells are terminally activated in DRlyp/lyp animals versus negatively regulated in DR+/+ animals; we have confirmed their functional role through preventing T1DM in rats with two different mast cell inhibiting drugs. Thus, we hypothesize that in addition to cytotoxic T effector cells, mast cells play a crucial early role in DRlyp/lyp diabetogenesis and in spite of their disease predisposition, TREG cells are sufficient to prevent T1DM in DR+/+ rats. We further hypothesize that associated with these states are distinct peripheral cytokine milieus, since we find the sera from human individuals at risk for T1DM or sera collected at onset induce a characteristic gene expression signature in healthy peripheral blood mononuclear cells (PBMC) that is distinct from normal controls and long standing T1DM. Importantly, we find observe this molecular signature as much as 5 years prior to T1DM onset. The predictable disease course of the BB rat enables longitudinal dissection of the activities of immune cell subpopulations and examining how these activities are manifest in the periphery. Thus we propose to 1) Temporally and spatially dissect the activities of antigen presenting, regulatory, and effector cells at defined timepoints through expression profiling and histological approaches. 2) The sera of WF, DR+/+ and DRlyp/lyp rats will be longitudinally evaluated for factors that induce gene expression in healthy PBMCs. Presence of candidate mediators and disease time point-specific biomarkers will be confirmed. Understanding the relationship between mast cells, T effector, and TREG cells in autoimmunity and has the potential to establish a new paradigm for immune regulation and create new insights to human disease.