Asthma, anxiety and GR abnormalities in non-human primates

 描述(由申请人提供)：我们先前证明，在反复的社会压力下，过敏原诱导的呼吸道炎症和高反应性(AHR)在暴露于重复的社会压力下的小鼠被增强。我们还发现，慢性社会扰乱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，应激可导致啮齿类动物脑细胞中GR基因异常甲基化，并修饰H3K4me3和H3K27me3。慢性心理社会应激对GR的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也不是最理想的。在加州大学戴维斯分校CNPRC研究的非人灵长类动物(猕猴)中，幼年猴子的抑制气质与2岁时皮质醇反应迟钝和AHR有关。我们推测，慢性应激猕猴哮喘易感性的增加与对内源性糖皮质激素的反应性受损有关。我们进一步假设，应激(表现为气质抑制)和过敏性哮喘(IgE升高、外周血嗜酸性粒细胞增多和AHR)的存在会增加NF-kB的活性，从而干扰免疫细胞中的GR。外显子1基因启动子的表观遗传修饰降低了应激动物GR的表达。目的1研究核因子-kB活性升高是否干扰慢性焦虑和哮喘患者外周血免疫细胞的GR表达、功能和类固醇反应。我们将比较这四组患者外周血单个核细胞对糖皮质激素的反应性、GR和NF-kB(一种促炎转录因子和GR抑制物)的表达、核转位和DNA结合。目的2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小鼠肺中GR基因表达减少。在啮齿类动物中也发现了应激诱导的NR3C1的DNA甲基化和组蛋白修饰。在猕猴外周血单个核细胞中，我们将利用DNA甲基化图谱和ChIPseq分析将GR异常表达与基因甲基化模式相关联。这些研究将为后续研究与人类疾病的相关性奠定基础，并为心理社会压力导致哮喘易感性的细胞-分子途径的机制研究奠定基础。

 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.