Estrogen receptor signaling, inflammation and ozone toxicity

项目摘要摘要 巨噬细胞通过调节肺部炎症的启动和消退来促进臭氧毒性； 这些过程由不同的巨噬细胞亚群介导，这些亚群被广泛归类为 促炎症/细胞毒(M1)和抗炎/伤口修复(M2)巨噬细胞。M1和M2 巨噬细胞的激活在一定程度上受细胞内代谢的控制；因此，虽然高糖酵解能力 与M1活性相关的是，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。新数据表明，臭氧毒性是由于M2激活受损所致 然而，未能消除炎症，其机制尚不清楚。白纹伊蚊的RNA-SEQ初步分析 臭氧暴露后收集的肺巨噬细胞显示雌激素受体显著丰富 差异表达基因间的信号通路。这与PPARγ的下调有关 表达，一种已知促进M2表型和炎症消退的转录因子 细胞内代谢向脂肪酸氧化转变。雌激素被证明通过以下方式调节PPARγ的表达 激活位于细胞膜的雌激素受体α(ESR1)，促进巨噬细胞抗 通过将新陈代谢转变为脂肪酸氧化的炎症活动；我们推测这一途径是重要的 巨噬细胞对臭氧的反应。我们假设臭氧干扰核外ESR1 巨噬细胞中的信号和PPARγ的下游激活；这导致脂肪酸受损 氧化和M2巨噬细胞激活导致炎症和 增加了组织损伤。为了检验这一假设，本文提出了三个目标。在前两个目标中(K99年期间 ，我们将分析臭氧对巨噬细胞生物能量学的影响以及ESR1信号的作用 巨噬细胞的免疫代谢和表型激活。目标3(R00奖励期)将重点放在1) 阐明臭氧干扰核外ESR1信号的机制，2)核外ESR1信号的作用 巨噬细胞中核ESR1的生物能量学和表型对臭氧的响应及其对肺的影响 3)通过抢救核外ESR1来减轻臭氧所致肺损伤的策略 发信号。这些研究结果将为巨噬细胞反应的潜在机制提供新的数据 这与共同小学导师的重点领域有很大的不同。这个 职业发展计划将在罗格斯大学进行，这是一所训练有素的一流机构 它将包括技术培训和专业发展活动。这些将提供 具备概念知识和培训以实现其成为一名 学术研究机构的资深研究员，负责指导独立研究计划 专注于阐明控制毒物引起的肺部疾病的细胞信号网络，并培训 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.