Project 4: Genetic Susceptibility

项目4：遗传易感性

• 批准号: 8309380
• 负责人: Clement Eugene Furlong
• 金额: $ 5.94万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8309380
• 关键词: Active Sites; Age; Animals; Archives; Biological Assay; Biological Markers; Blood; Butyrylcholinesterase; Child; Child health care; Chlorpyrifos; Communities; Data; Detection; Development; Dose; Drops; Environmental Exposure; Environmental Health; Erythrocytes; Exposure to; Fetus; Genes; Genetic; Genetic Predisposition to Disease; Genomics; Goals; Hour; Individual; Knock-out; Label; Link; Liquid Chromatography; Mass Spectrum Analysis; Measurement; Measures; Modification; Molecular; Monitor; Mus; National Children' s Study; O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; Paraoxonase 1; Parents; Pathway interactions; Peptide Hydrolases; Peptide antibodies; Peptides; Pesticides; Plasma; Predisposition; Proteins; Proteomics; Protocols documentation; Reaction; Research; Research Project Grants; Resources; Risk; Role; Running; Sampling; Sensitivity and Specificity; Serine; Specificity; System; Technology; Testing; Time; Tissues; Toxicokinetics; adduct; aryldialkylphosphatase; base; community based participatory research; critical period; design; esterase inhibitor; field study; gene environment interaction; in vivo; meetings; mouse model; neonatal exposure; phosphorothioate; postnatal; pregnant; pup; research study; response; urinary

The proposed studies aim to develop high throughput protocols for assessing OP exposures by characterizing specific biomarker proteins in blood, and to make use of these more accurate measures of exposure to investigate gene/environment interactions related to genetic variability in the paraoxonase (PONI) gene, particularly with respect to OP exposures that occur during early development. Specific Aim 1 is to validate high throughput immunomagnetic bead (1MB) based isolation protocols for rapid purification and analysis of the active site peptides of two proteins, red cell acyl peptide hydrolase (APH) and plasma butyrylcholinesterase (BChE), as biomarkers of exposure to OP compounds. A targeted proteomics strategy based on microcapillary liquid chromatography-selected reaction monitoring-mass spectrometry (¿mu¿LC-SRM-MS) will be validated and used to quantify the percentage modification of BChE and APH in archived samples from the Community Based Participatory Research Project. Data from the biomarker experiments will be correlated with other data available on urinary metabolite levels, inhibition of APH and/or BChE, and PONI status. Specific Aims 2 and 3 use knockout and humanized mouse models to determine the importance of PONI status for OP exposures that occur during critical periods of early development. Specific Aim 2 is to determine the extent to which APH and BChE are covalently modified following test exposure of pregnant mice to chlorpyrifos (CP) or CPO. Specific Aim 3 is to determine the extent to which BChE and APH are covalently modified following exposure of neonatal mice to CP or CPO. These aims will evaluate interactions among biomarkers of sensitivity, exposure and response in an in vivo system, allowing us to determine full dose responses of the OP compounds and generate quantitative biomarker data.

拟议的研究旨在制定高吞吐量协议，以通过表征来评估OP暴露 血液中的特定生物标志物蛋白，并利用这些更准确的暴露措施 研究与二氧化二氧酶（PONI）基因遗传变异性有关的基因/环境相互作用， 特别是关于早期开发过程中发生的OP暴露。具体目标1是验证高 基于吞吐量免疫磁珠（1MB）的隔离方案，用于快速纯化和分析 两种蛋白质的活性位点胡椒粉，红细胞酰基肽水解酶（APH）和血浆丁酰胆碱酯酶 （BCHE），作为暴露于OP化合物的生物标志物。基于微毛细血管的靶向蛋白质组学策略 液相色谱选择反应监测质量光谱法（MU¿LC-SRM-MS）将经过验证，并将被验证 用于量化基于社区的存档样本中BCHE和APH的修改百分比 参与性研究项目。来自生物标志物实验的数据将与其他数据相关 可在尿中代谢物水平，抑制APH和/或BCHE以及PONI状态可用。具体目标2和3 使用敲除和人源化的鼠标模型来确定PONI状态对OP暴露的重要性 发生在早期发展的关键时期。具体目标2是确定APH和 在怀孕小鼠暴露于毒死rif（CP）或CPO之后，BCHE经过共价修改。具体目标 3是确定新生小鼠暴露后BCHE和APH共价修改的程度 到CP或CPO。这些目标将评估敏感性，暴露和反应的生物标志物之间的相互作用 一个体内系统，使我们能够确定OP化合物的全剂量反应并产生定量 生物标志物数据。