Investigating gene x environment interaction using human exposures to O3 & LPS

描述（由申请人提供）：本NIEHS挑战基金是针对挑战主题04-ES-102：使用受控的人类暴露调查基因与环境的相互作用而编写的。正如RFA所述，“仔细控制人类受试者暴露于低水平环境毒物，如环境颗粒物，臭氧或柴油废气，提供了一个机会，帮助增加动物研究和基于人群的研究，以更好地了解遗传和暴露（GxE）的相互作用。”为了解决这个问题，我们将采用两种经批准的暴露方案来获取个体在0.4 ppm臭氧下持续2小时的反应表型，并吸入20,000单位的内毒素。我们将通过招募志愿者加入臭氧协议，然后邀请完成臭氧研究的人进行内毒素挑战来实现这一目标。臭氧是一种常见的污染物，而内毒素是颗粒物和生物气溶胶的主要成分。我们将检查的应答表型包括肺功能、气道炎症、气道和循环单核细胞、巨噬细胞和树突状细胞的免疫表型、气道和循环吞噬细胞功能（氧化破裂和磷酸化酶的吞噬）和介质分泌。招募最初将集中在谷胱甘肽- s转移酶M1 （GSTM1）基因型和烟酰胺腺嘌呤二核苷酸（磷酸）还原：醌氧化还原酶（NQO1）基因型的零和充足和单核苷酸多态性（snp），以及由此产生的单倍型。其他常见的单核苷酸多态性（snp）存在于NF-E2、相关因子2 （NRF2）、GSTP1、GSTT1、谷胱甘肽过氧化物酶（GPX1）、血红素加氧酶-1 （HO-1）和超氧化酶歧化酶2 （SOD2）、cd14基因启动子TLR2、TLR4；IL-1， IL-1R和TNF基因启动子。虽然招募主要集中在GSTM1和NQO1上，但其他基因的等位基因频率足够高，因此也可以评估这些snp的影响。利用从气道和循环细胞中回收的mRNA进行基因表达谱分析也将用于鉴定调节对臭氧和LPS反应的新的候选基因。

DESCRIPTION (provided by applicant): This NIEHS challenge grant is written in response to challenge topic 04-ES-102: Investigating gene x environment interaction using controlled human exposures. As stated in the RFA, "Carefully controlled exposures of human subjects to low levels of environmental toxicants, such as ambient particulate matter, ozone, or diesel exhaust, provide an opportunity to help augment animal studies and population-based studies to better understand the interaction of genetics and exposure (GxE). To address this topic, we will employ two approved exposure protocols to access response phenotypes of individuals to 0.4 ppm ozone for 2 hours, and inhalation of a 20,000 units of endotoxin. We will achieve this by recruiting volunteers into the ozone protocol, and then inviting those who complete the ozone study to undertake endotoxin challenge. Ozone is a commonly occurring pollutant, and endotoxin is a major component of particulate matter and bioaersols. The response phenotypes we will examine include lung function, airway inflammation, immunophenotyping of airway and circulating monocytes, macrophages and dendritic cells, airway and circulating phagocyte function (oxidative burst and phagocytosis of opsonized zymosan) and mediator secretion. Recruitment will initially be focused on the null and sufficient glutathione-S-transferase M1 (GSTM1) genotype and null vs. sufficient and single nucleotide polymorphisms (SNPs) for nicotinamide adenine dinucleotide (phosphate) reduced:quinone oxidoreductase (NQO1), with the resulting haplotypes. Other common single nucleotide polymorphisms (SNPs) of interest exist in NF-E2, related factor 2 (NRF2), GSTP1, GSTT1, glutathione peroxdiase (GPX1), Heme oxygenase-1 (HO-1), and Superoxidase dismutase 2 (SOD2), theCD14 gene promoter, TLR2, TLR4; IL-1 , IL-1R, and the TNF gene promoter. While recruitment is focused on GSTM1 and NQO1, allele frequencies of the other genes are high enough that assessment of the effect of these SNPs will be achieved as well. Gene expression profiling using mRNA recovered from airway and circulating cells will also be conducted to identify novel candidate genes that regulate response to ozone and LPS. PUBLIC HEALTH RELEVANCE: Knowledge obtained from this research can be used for mitigation of the effect of environmental pollutants on persons with lung and cardiovascular disease through development of personal mitigation strategies and policymaking. The Clean Air Act requires that the National Ambient Air Quality Standards protect vulnerable populations, including those which genetically define vulnerability. Finally, these studies will also add to fundamental advances in our understanding of the pathophysiology of common human conditions.