Project 1: Biomarkers of Susceptibility to Environmentally-Induced Diseases

对氧磷酶家族(PON1、PON2和PONS)位于人类7号染色体上。与高密度脂蛋白相关的PON1代谢有机磷(OP)化合物、氧化脂质、药物和群体感应因子。PON1基因多态性影响催化效率和血浆水平。两个PON2编码区多态的后果尚不清楚。PON1在肝脏中合成并分泌到血浆中，而PON2在包括脑在内的组织中普遍表达。PON_2不能水解有机磷，但具有很强的抗氧化性。与男性对照组相比，男性帕金森氏症患者的PON1状态发生了变化。建议的研究旨在增加我们对PON1、PON2和PONS功能的了解，以确定环境诱导的神经毒性和神经退行性疾病的易感性。具体目标1继续我们关于华盛顿州农药操作者队列中PON1状态与OP毒性易感性的关系的研究。除了监测血浆胆碱酯酶水平外，我们还将包括一种新的定量质谱仪(MS)分析 蛋白质生物标记物因暴露在OP操作者和航空事故中而改变。特定目的2研究PON1和羧酸酯酶在调节杀虫剂混剂毒性中的相互作用。具体目的3研究PON1在毒死蜱毒杀的产前发育神经毒性中的作用，其中假设母鼠的PON1状态对胎儿保护很重要。这些研究包括PON1缺失小鼠和人源化PON1转基因小鼠(tgHuPON1-Q192和tgHuPON1-R192)，包括生化、行为学、组织病理学和分子终点。特定目标4调查桥1、2的作用 3和高密度脂蛋白在帕金森病(PD)中的组成。我们之前的研究表明，与对照组相比，男性帕金森病患者的PON1状态不同，作为后续研究，我们将使用基于MS的蛋白质组学方法来检查帕金森病受试者和匹配对照组的高密度脂蛋白的组成。缺乏PON蛋白的小鼠也将接受对多巴胺能神经毒素MPTP的敏感性测试。具体目标5涉及与项目5的合作，在植物系统中表达稳定的兔子PON1，以补救OP泄漏。

The paraoxonase family of genes (PON1, PON2 and PONS) is located on human chromosome 7. HDL-associated PON1 metabolizes organophosphorus (OP) compounds, oxidized lipids, drugs and quorum-sensing factors. PON1 polymorphisms affect catalytic efficiency and plasma levels. The consequences of the two PON2 coding region polymorphisms are unknown. PON1 is synthesized in liver and secreted into plasma, whereas PON2 is ubiquitously expressed in tissues, including brain. PON2 does not hydrolyze OPs but has strong antioxidant properties. PON1 status was found to be altered in male Parkinson's patients compared with male control subjects. The proposed studies aim to increase our knowledge of PON1, PON2 and PONS functions in determining susceptibility to environmentally-induced neurotoxicity and neurodegenerative diseases. Specific Aim 1 follows up on our studies on the relationship of PON1 status and susceptibility to OP toxicity in a cohort of Washington State pesticide handlers. In addition to monitoring plasma cholinesterase levels, we will include a novel quantitative mass spectrometric (MS) analysis of protein biomarkers modified by exposures in OP handlers and airline incidents. Specific Aim 2 investigates the interplay of PON1 and carboxylesterase in modulating the toxicity of insecticide mixtures. Specific Aim 3 examines the role of PON1 in modulating prenatal developmental neurotoxicity of chlorpyrifos oxon, where PON1 status of dams is hypothesized to be important for fetal protection. These studies with PON1 null mice and humanized PON1 transgenic mice (tgHuPON1-Q192 & tgHuPON1-R192) include biochemical, behavioral, histopathological and molecular end-points. Specific Aim 4 investigates the role of PONs 1, 2 and 3 and HDL composition in Parkinson's disease (PD). As a follow-up on our previous studies that showed differences in PON1 status in male PD patients compared with controls, we will use an MS-based proteomic approach to examine HDL composition in PD subjects vs. matched controls. Mice lacking PON proteins will also be tested for sensitivity to the dopaminergic neurotoxin MPTP. Specific Aim 5 involves a collaboration with Project 5 to express stable rabbit PON1 in plant systems for remediating OP spills.