Early life ENM-exposure and the impact on neurobehavioral and cardiovascular outcomes and biochemical mechanisms

项目摘要 本意见书是根据RFA-ES-15-013号文件编写的，该文件将建立一个 环境健康科学-纳米技术健康影响研究(NHIR)联盟： 工程纳米材料与生物系统相互作用的综合评价 U01。Nhir联盟将合作研究ENM和生物之间的相互作用 提供ENM暴露的全面反应概况的系统，以指导良性ENM的发展 并支持安全和可持续地使用纳米技术。这项建议的目的是调查 该联盟选择了具有广泛物理化学性质的ENM，以揭示ENM的早期生命 暴露会影响神经行为和心血管结果，并导致生化变化。这个 发育中的新生儿有不成熟的胃肠功能，具有发育中的屏障功能，这可能允许 比成人对ENM的摄取更多。接触化学物质会影响发育中的大脑 无论是直接还是间接的。心血管系统也被证明对 新能源管理体系的影响。 该项目将包括三个相互关联的调查领域，目的如下： 1.体外研究ENMS对肠屏障完整性和通透性的影响。我们 将在两个成熟的小肠模型中测试联盟选择的ENM，使用1) 肠细胞和杯状细胞；2)肠细胞和M细胞。 2.测定新生大鼠和幼年大鼠ENM的组织分布和细胞定位 神经行为和心血管影响的测定。对新城疫的生物反应将是 新生及幼年雌性和雄性大鼠出生后每日口服ENMS的实验研究 (PND)2-20，并于PND 4、9和21实施安乐死。ENM组织分布的综合评估， 功能性神经行为观察，心血管测量，神经递质测量，以及 非靶向和靶向代谢组学分析(AIM 3)将在PND 4、9和21日进行。 3.表征血清、脑组织和盲肠内容物的生化特征 与ENM暴露以及神经学和心血管评估有关。研究将集中在 确定ENMS如何扰乱男性血清、脑组织和盲肠内容物的生化特征 和雌性幼鼠。评估将包括使用核磁共振代谢组学，一个16通道的ECD库仑阵列来 同时对20种神经递质进行定量，并对188种化合物进行定量检测，包括 酰基肉碱、氨基酸、生物胺、己糖、甘油磷脂、磷脂酰胆碱和 神经鞘脂类。

Project Summary This submission is prepared in response to RFA-ES-15-013, which will establish a National Institute of Environmental Health Sciences - Nanotechnology Health Implications Research (NHIR) Consortium: Comprehensive Evaluation of Interactions Between Engineered Nanomaterials (ENMs) and Biological System U01s. The NHIR Consortium will collaborate to investigate the interaction between ENMs and biological systems to provide comprehensive response profiles of ENM exposures to guide development of benign ENMs and support safe and sustainable use of nanotechnology. The aim of this proposal is to investigate Consortium-selected ENMs, with a broad range of physicochemical properties, to reveal how early life ENM exposure impacts neurobehavioral and cardiovascular outcomes and cause changes in biochemistry. The developing neonate has immature gastrointestinal function, with a developing barrier function, that may allow greater uptake of ENMs than in an adult. The developing brain can be impacted by exposure to chemicals both directly and indirectly. The cardiovascular system has also been shown to be particularly sensitive to the effects of ENMs. The project will consist of three inter-related areas of investigation with the following aims: 1. To investigate the impact of ENMs on the integrity and permeability of the intestinal barrier in vitro. We will test the Consortium-selected ENMs in two well-established models of the small intestine using 1) enterocytes and goblet cells, and 2) enterocytes and M cells. 2. To determine the ENM tissue distribution and cellular localization in exposed neonatal and juvenile rats with determination of neurobehavioral and cardiovascular effects. The biological response to ENMs will be studied in female and male neonatal and juvenile rats with daily ENMs administered orally from postnatal day (PND) 2-20, and euthanized at PND 4, 9, and 21. Comprehensive assessment of the ENM tissue distribution, functional neurobehavioral observations, cardiovascular measurements, neurotransmitter measurement, and untargeted and targeted metabolomics analysis (aim 3) will be conducted on PND 4, 9 and 21. 3. To characterize perturbations in biochemical profiles of serum, brain tissue, and cecal contents associated with ENM exposure, and with neurological and cardiovascular assessments. Studies will focus on determining how ENMs perturb the biochemical profiles of the serum, brain tissue, and cecal contents of male and female rat pups. Assessments will include the use of NMR metabolomics, a 16 channel ECD CoulArray to simultaneously quantitate 20 neurotransmitters, and a quantitative assay for 188 compounds including acylcarnitines, amino acids, biogenic amines, hexose, glycerophospholipids, phosphatidylcholines, and sphingolipids.