Activation of Nrf2 during embryonic development - mechanisms and consequences

摘要 生命早期接触有毒物质会导致胰岛畸形，这可能使个人容易患上 糖尿病。谷胱甘肽氧化还原微环境在胚胎发育和细胞发育中起着重要作用 信号，其扰动会导致功能或结构的改变，只有在 随后的压力或年龄。令人惊讶的是，人们对胚胎如何应对氧化应激知之甚少 毒物暴露对胰腺β细胞发育的影响。该项目采用多层次的方法，使用 最先进的技术来阐明接触辐射的复杂病理生理机制 全氟和多氟烷基物质(PFAS)会引起氧化应激，使胰岛发育脱轨， 对β-CELL函数的影响。我们检验了中心假设，即偏离GSH氧化还原 微环境与转录因子Nrf2在错误位置和错误位置的异常激活 时间-损害β细胞的发育和功能。这个项目有三个首要目标：1)深化 我们对NRF2激活在胚胎β细胞和胰岛发育中的作用的理解；2)确定 全氟辛烷磺酸对胰岛素生物合成的影响；以及3)确定β细胞的脆性和生命后期代谢的生物指标 可以转化为对人类健康的影响。我们将使用转基因斑马鱼、共聚焦显微镜和 免疫荧光、氧化还原蛋白质组学和胰岛素错误折叠分析，以及培养的β细胞 暴露于两种常见的全氟辛烷磺酸(全氟辛烷磺酸、全氟辛烷磺酸)和一种传统的水性成膜泡沫(AFFF)。这部作品 将对发育毒理学、氧化还原生物学和 健康和疾病的发展起源，并为发展以科学为基础的 PFAS指南、临床干预目标和公共卫生政策。

Abstract Early life stage exposures to toxicants can result in islet malformations, which may predispose individuals to diabetes. The glutathione redox microenvironment plays fundamental roles in embryonic development and cell signaling, perturbation of which can result in functional or structural alterations that only become apparent with subsequent stress or age. Surprisingly little is known about how embryos respond to oxidative stress, or the impact of toxicant exposures on pancreatic β-cell development. This project takes a multi-level approach using state-of-the-art techniques to elucidate the complex pathophysiological mechanisms by which exposures to Per-and-polyfluoroalkyl substances (PFAS) that cause oxidative stress derail islet development, and the consequences for β-cell function. We test the central hypothesis that deviations from the GSH redox microenvironment and aberrant activation of the transcription factor Nrf2- at the wrong place and the wrong time- impair β-cell development and function. There are three overarching goals of this project: 1) to deepen our understanding of the role of Nrf2 activation in embryonic β-cells and islet development; 2) ascertain the impact of PFAS on insulin biosynthesis; and 3) identify β-cell fragility and bioindicators of later-life metabolic impacts that can be translated to human health. We will use transgenic zebrafish, confocal microscopy and immunofluorescence, redox proteomics and insulin misfolding assays, and cultured β-cells to investigate exposures to two common PFAS (PFOS, PFHxS), and a legacy aqueous film-forming foam (AFFF). This work will have a sustained and powerful impact on the fields of developmental toxicology, redox biology, and the developmental origins of health and disease and provides critical advances towards developing science-based PFAS guidelines, targets for clinical interventions, and public health policies.