Activation of Nrf2 during embryonic development - mechanisms and consequences

摘要 生命早期暴露于有毒物质可导致胰岛畸形，这可能使个体易患 糖尿病谷胱甘肽氧化还原微环境在胚胎发育和细胞生长中起着重要作用 信号传导，其扰动可导致功能或结构改变，这些改变只有在 随后的压力或年龄。令人惊讶的是，关于胚胎如何应对氧化应激，或者 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， 国家的最先进的技术，以阐明复杂的病理生理机制， 引起氧化应激的全氟烷基和多氟烷基物质（PFAS）破坏胰岛发育， 影响β细胞功能。我们测试的中心假设，从谷胱甘肽氧化还原偏差 微环境和转录因子Nrf 2的异常激活-在错误的地方和错误的 时间-损害β细胞发育和功能。该项目有三个总体目标：1）深化 我们对Nrf 2激活在胚胎β细胞和胰岛发育中的作用的理解; 2）确定Nrf 2激活在胚胎β细胞和胰岛发育中的作用。 PFAS对胰岛素生物合成影响;以及3）鉴定β细胞脆性和晚年代谢的生物指标 这些影响可以转化为人类健康。我们将使用转基因斑马鱼，共聚焦显微镜， 免疫荧光，氧化还原蛋白质组学和胰岛素错误折叠测定，以及培养的β细胞来研究 暴露于两种常见的全氟辛烷磺酸（PFOS，PFHxS）和一种传统的水性成膜泡沫（AFFF）。这项工作 将对发育毒理学、氧化还原生物学和生物学领域产生持续而强大的影响。 健康和疾病的发展根源，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.