Estrogen receptor signaling, inflammation and ozone toxicity

项目摘要 大环内酯通过调节肺部炎症的发生和消退而促进臭氧毒性; 这些过程由不同的巨噬细胞亚群介导， 促炎/细胞毒性（M1）和抗炎/伤口修复（M2）巨噬细胞。m1和m2 巨噬细胞的活化部分受细胞内代谢控制;因此，虽然高糖酵解能力是 与M1活性相关，脂肪酸氧化和线粒体氧化磷酸化增加， M2巨噬细胞激活所需的。新的数据表明，臭氧毒性是由于受损的M2激活 以及不能解决炎症，然而，其机制尚不清楚。初步RNA-seq分析 臭氧暴露后收集的肺巨噬细胞显示雌激素受体的显着富集 差异表达基因之间的信号通路。这与PPAR γ表达下调有关 表达，一种转录因子，已知通过转移促进M2表型和炎症消退 细胞内代谢到脂肪酸氧化。雌激素已被证明可以通过以下方式调节PPAR γ的表达： 激活位于细胞膜上的雌激素受体α（ESR1），并促进巨噬细胞抗- 炎症活动通过转移代谢脂肪酸氧化;我们推测这一途径是重要的 对臭氧的反应。我们假设臭氧干扰核外ESR1 巨噬细胞中的信号传导和PPAR γ的下游活化;这导致脂肪酸受损 氧化和M2巨噬细胞活化导致炎症的异常消退， 增加组织损伤。提出了三个目标来检验这一假设。在前两个目标中（K99时期）， 我们将分析臭氧对巨噬细胞生物能量学的影响以及ESR1信号传导的作用 在巨噬细胞免疫代谢和表型活化中的作用。目标3（R00奖励期）将侧重于1） 阐明臭氧干扰细胞核外ESR1信号传导的机制，2）细胞核外 核ESR1在巨噬细胞生物能量学和表型响应臭氧和如何影响肺 损伤，以及3）制定策略，通过挽救核外ESR1来减轻臭氧引起的肺损伤 发信号。这些研究结果将为巨噬细胞反应机制提供新的数据 吸入臭氧，并代表了一个重大偏离共同主要导师的重点领域。的 职业发展计划将在罗格斯大学，一个一流的机构，具有强大的培训执行 环境的影响，并将包括技术培训和专业发展活动。这些将提供 候选人的概念知识和培训所需的实现他的长期目标，成为一个 在学术研究机构指导独立研究项目的资深研究员 专注于阐明细胞信号网络控制毒物引起的肺部疾病和培训， 下一代环境健康科学家

Project Summary Abstract Macrophages contribute to ozone toxicity by regulating both the initiation and resolution of lung inflammation; these processes are mediated by distinct macrophage subpopulations broadly classified as proinflammatory/cytotoxic (M1) and anti-inflammatory/wound repair (M2) macrophages. M1 and M2 macrophage activation is controlled, in part, by intracellular metabolism; thus, while high glycolytic capacity is associated with M1 activity, increases in fatty acid oxidation and mitochondrial oxidative phosphorylation are required for M2 macrophage activation. New data suggest that ozone toxicity is due to impaired M2 activation and a failure to resolve inflammation, however, mechanisms are not known. A preliminary RNA-seq analysis of lung macrophages collected after ozone exposure revealed significant enrichment of the estrogen receptor signaling pathway among differentially expressed genes. This was associated with down-regulation of PPARγ expression, a transcription factor known to promote M2 phenotype and resolution of inflammation by shifting intracellular metabolism to fatty acid oxidation. Estrogen has been shown to regulate PPARγ expression by activating estrogen receptor alpha (ESR1) located at the cell membrane, and to promote macrophage anti- inflammatory activity by shifting metabolism to fatty acid oxidation; we speculate that this pathway is important in macrophage responses to ozone. We hypothesize that ozone interferes with extra-nuclear ESR1 signaling in macrophages and downstream activation of PPARγ; this leads to impaired fatty acid oxidation and M2 macrophage activation resulting in aberrant resolution of inflammation and increased tissue injury. Three aims are proposed to test this hypothesis. In the first two aims (K99 period of this award), we will analyze the effects of ozone on macrophage bioenergetics and the role for ESR1 signaling in macrophage immunometabolism and phenotypic activation. Aim 3 (R00 award period) will focus on 1) elucidation of mechanisms by which ozone interferes with extra-nuclear ESR1 signaling, 2) the role of extra- nuclear ESR1 in macrophage bioenergetics and phenotype in response to ozone and how this influences lung injury, and 3) developing strategies to mitigate ozone-induced lung injury by rescuing extra-nuclear ESR1 signaling. Results of these studies will provide novel data on mechanisms underlying macrophage responses to inhaled ozone and represent a significant departure from the co-primary mentor’s areas of emphasis. The career development plan will be performed at Rutgers University, a first-class institution with a strong training environment and will consist of technical training and professional development activities. These will provide the candidate with conceptual knowledge and training required to achieve his long-term goal of becoming an established investigator at an academic research institution directing an independent research program focused on elucidating cellular signaling networks controlling toxicant-induced lung disease and training the next generation of environmental health scientists.