Asthma, anxiety and GR abnormalities in non-human primates

 描述（由申请人提供）：我们先前证明，在暴露于反复社会压力的小鼠中，过敏原诱导的气道炎症和高反应性（AHR）增强。我们还发现，小鼠的慢性社会破坏应激导致免疫细胞和肺中糖皮质激素受体（GR）的表达和功能受损。其他研究人员发现，应激可诱导啮齿类动物脑细胞中GR基因和修饰的H3 K4 me 3和H3 K27 me 3的异常甲基化模式。慢性心理社会应激对GR影响的意义和机制尚不清楚，目前可用的啮齿动物慢性心理社会应激模型不太理想。在加州大学戴维斯分校的CNPRC研究的非人灵长类动物（恒河猴）中，幼年猴的抑制性气质与2岁时皮质醇反应迟钝和AHR相关。我们推测，慢性应激恒河猴哮喘易感性增高与内源性糖皮质激素反应性受损有关。我们进一步假设，存在的压力（表现为抑制气质）和过敏性哮喘（IgE升高，外周血嗜酸性粒细胞增多症和AHR）增加NF-κ B的活性，干扰免疫细胞中的GR。外显子1基因启动子的表观遗传修饰减少应激动物中GR的表达。目的1：研究慢性焦虑和哮喘灵长类动物外周血免疫细胞NF-κ B活性升高是否干扰GR的表达、功能和激素反应性。我们将比较糖皮质激素的反应性和表达的GR和NF-κ B（促炎转录因子和GR抑制剂），核转位和DNA结合在外周血单核细胞从这四个组。目的二是研究表观遗传学变化在灵长类动物应激和AHR免疫细胞GR表达调控中的作用。我们先前发现应激诱导的小鼠肺GR mRNA表达减少。在啮齿类动物中也发现了应激诱导的NR 3C 1 DNA甲基化和组蛋白修饰。在来自恒河猴的外周血单核细胞中，我们将使用DNA甲基化作图和ChIPseq分析将异常GR表达与基因甲基化模式相关联。这些研究将为随后的研究奠定基础，以人类疾病的相关性和机制研究的细胞分子途径，导致哮喘易感性，由于心理社会压力。

 DESCRIPTION (provided by applicant): We previously demonstrated that allergen-induced airway inflammation and hyper responsiveness (AHR) was enhanced in mice exposed to repeated social stress. We also found that chronic social disruption stress in mice caused impaired glucocorticoid receptor (GR) expression and function in immune cells and in the lung. By other investigators stress was shown to induce abnormal methylation pattern of the GR gene and modified H3K4me3 and H3K27me3 in brain cells of rodents. The significance and mechanisms of the chronic psychosocial stress effects on the GR remain unclear and the currently available chronic models of psychosocial stress in rodents are less than optimal. In non-human primates (rhesus macaques) studied at the CNPRC at UC Davis, inhibited temperament in infant monkeys was associated with blunted cortisol responsiveness and AHR at 2 years of age. We postulate that a heightened predisposition to asthma of chronically stressed rhesus macaques is related to an impaired responsiveness to endogenous glucocorticoids. We further hypothesize that presence of stress (manifested as inhibited temperament) and allergic asthma (elevated IgE, peripheral blood eosinophilia and AHR) increases NF-kB activity that interferes with the GR in immune cells. Epigenetic modifications of the Exon 1 gene promoter reduce GR expression in stressed animals. Aim 1 is to study whether increased NF-kB activity interferes with GR expression, function and steroid responsiveness of peripheral blood immune cells from primates that exhibit chronic anxiety and asthma. We will compare glucocorticoid responsiveness and expression of GR and NF-kB (a pro-inflammatory transcription factor and GR inhibitor), nuclear translocation and DNA binding in peripheral blood mononuclear cells from these four groups. Aim 2 is to study the role of epigenetic changes in regulation of GR expression in immune cells of primates that exhibit stress and AHR. We previously found stress-induced reduction in GR mRNA expression in the lung of mice. Stress-induced DNA methylation of NR3C1 and histone modifications was also identified in rodents. In peripheral blood mononuclear cells from rhesus macaques we will correlate abnormal GR expression with gene methylation patterns using DNA methylation mapping and ChIPseq analysis. These studies will lay the groundwork for subsequent investigations into the relevance to human disease and mechanistic studies on the cellular-molecular pathways that lead to asthma predisposition due to psychosocial stress.