Dissection of cellular interactions in T1DM with integrated functional genomics

项目总结 认识到系统方法对复杂疾病的力量，在最初的筹资期间，我们 开发了一个高度受控的三色阵列和分析平台，专注于质量数据的采集。 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 生物育种(BB)大鼠1型糖尿病(T1 DM)模型。BB-DRlyp/LYP大鼠自发性糖尿病 由于Gimap5基因突变，导致他们缺乏调节性T(Treg)细胞。DR/大鼠 拥有完整的Gimap5基因，不会发展为自发性T1 DM，但在耗尽时会患上糖尿病 特雷格细胞。BB大鼠的T1 DM还涉及一种细胞毒性T细胞，因为它们的耗尽是保护性的。我们的基因 糖尿病前期胰腺淋巴结(PLN)的表达研究表明，肥大细胞是终末的 在DRlyp/LYP动物中激活，而在DR/动物中负调控；我们已经证实了他们的 通过两种不同肥大细胞抑制药预防大鼠T1 DM的作用。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中起着关键的早期作用 糖尿病的发生和尽管其疾病易感性，Treg细胞足以预防DR/T1 DM 老鼠。我们进一步假设，与这些状态相关的是不同的细胞因子环境，因为我们发现 T1 DM高危人群或新近发病的T1 DM患者血清与健康人的孵育 外周血单个核细胞(PBMC)诱导一种独特的基因表达特征 来自正常对照组和长期存在的T1 DM。重要的是，我们发现观察这种分子特征 在T1 DM发病前5年。BB大鼠可预测的病程使其能够纵向解剖 免疫细胞亚群的活动，并研究这些活动是如何在 外围设备。因此，我们建议1)在时间和空间上剖析抗原提呈的活动， 通过基因表达谱和组织学研究确定发病前时间点的调节细胞和效应细胞 接近了。2)WF、DR/和DRlyp/LYP大鼠的血清将被纵向评估以下因素 诱导健康外周血单核细胞基因表达。候选介体的存在和疾病时间点特定 生物标志物将得到确认。了解肥大细胞、T效应细胞和Treg细胞之间的关系 自身免疫，有可能建立免疫调节的新范式，并创造新的见解 人类疾病。

PROJECT SUMMARY 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 a 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 cytokine milieus, since we find incubation of sera derived from human individuals at risk for T1DM or recent onset T1DM with healthy peripheral blood mononuclear cells (PBMC) induce a characteristic gene expression signature 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 manifested in the periphery. Thus we propose to 1) temporally and spatially dissect the activities of antigen presenting, regulatory, and effector cells at defined pre-onset timepoints through gene 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.