Gene-Environment Interactions with Ozone and Non-atopic Asthma

基因-环境与臭氧和非特应性哮喘的相互作用

基本信息

批准号：
10302827
负责人：
Samir Kelada
金额：
$ 24.42万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10302827
关键词：
Address Air Air Pollutants Alleles Applications Grants Asthma Attenuated Backcrossings Bioinformatics Biological Bronchoalveolar Lavage Fluid Candidate Disease Gene Cell Count Cells Child Climate Clinical Research Critiques Data Development Endotoxins Environment Eosinophilia Epidemiology Epithelial Epithelial Cells Exhibits Exposure to Future Gene Expression Genes Genetic Genetic Models Genotype Goals Hour House Dust Mite Allergens Human Hyperplasia Immunity Immunologics Immunologist Inbred Strains Mice Individual Injury Interleukin-13 Interleukin-5 Investigation Laboratory Study Leukocytes Lung Lung diseases Lymphoid Cell Maps Mediating Metaplastic Cell Molecular Mucous body substance Mus Ozone Pathway interactions Phenotype Population Production Quantitative Trait Loci Rattus Recombinant Inbred Strain Research Personnel Signal Transduction Specificity Surveys TSLP gene Testing Work adverse outcome airway epithelium airway hyperresponsiveness airway inflammation asthma exacerbation base cigarette smoke cytokine density eosinophil eosinophilic inflammation epidemiology study experimental study gene environment interaction genetic approach genetic variant genomic locus innate immune mechanisms innovation insight mRNA Expression nonhuman primate novel ozone exposure response single-cell RNA sequencing targeted treatment whole genome

项目摘要

PROJECT SUMMARY Epidemiologic studies have provided evidence that ozone (O3) exposure is associated with the development of asthma, in particular the non-atopic (non-allergic) sub-type. These findings are supported by clinical studies showing that ambient O3 exposure is associated with eosinophilic airway inflammation in non-atopic children. Laboratory studies have shown that O3 exposure causes mucous cell metaplasia/hyperplasia in non-human primates, rats and mice. This includes recent results from our groups demonstrating that classical inbred strains of mice exposed to 0.8 ppm O3 (4 hours/day) for 9 days develop eosinophilic airway inflammation in addition to mucous cell hyperplasia. Thus, data in three species and humans provide strong rationale for studies geared towards understanding the underlying mechanisms of the O3 ® non-atopic asthma association. We take a genetic approach to the question of mechanism. We surveyed strains from the Collaborative Cross (CC) mouse population, a new panel of recombinant inbred strains and identified one strain, CC002, that exhibited much higher levels of eosinophilic inflammation after repeated O3 exposure than any other strain tested to date. O3 exposed CC002 mice also developed mucous cell metaplasia and airway hyper- responsiveness (AHR), two other hallmark phenotypes of asthma. Thus, CC002 provides a new genetic model that we can exploit to identify mechanisms underlying the of O3-induced non-atopic asthma association. In the first Aim, we will test the hypothesis that CC002’s exaggerated response to O3 exposure are due to dysregulated epithelial signaling via alarmins that in turn stimulates type 2 innate lymphoid cells (ILC2s). We will determine if epithelial-derived innate cytokines (IL-1a/b, IL-25, IL-33, TSLP) are increased in O3-exposed CC002 mice compared to non-responder strains, and if neutralization of these cytokines attenuates O3-induced airway inflammation and AHR. Next, we will determine if ILC2 numbers and activation are increased in O3- exposed CC002 mice, and if depletion of ILC2s or inhibition of ILC2-derived cytokines (IL-5, IL-13) ameliorates O3-induced airway inflammation and AHR in CC002 mice. To comprehensively characterize molecular responses to O3 in epithelia, ILC2s, and other leukocytes, we will perform single cell RNA-sequencing. Using this innovative and powerful approach, we will quantify differences in gene expression and cellular composition +/- O3, both within and between strains. We will also examine the specificity of CC002’s response by testing whether this strain exhibits exaggerated responses to other exposures, including endotoxin, cigarette smoke, and house dust mite allergen. In the second aim, we will identify the genetic loci that render CC002 sensitive to O3. We will perform quantitative trait locus mapping to identify chromosomal regions harboring CC002 alleles associated with O3 sensitivity, followed by bioinformatic analyses to identify high priority candidate genes at each locus. In total, our work will reveal mechanisms underlying the association between O3 exposure and the development of non-atopic asthma phenotypes and identify potential pathways to target therapeutically.

项目摘要流行病学研究提供了证据，表明臭氧（O3）暴露与哮喘，尤其是非原子（非过敏性）亚型。这些发现得到了临床研究的支持表明环境O3暴露与非原子儿童的嗜酸性气道注射有关。实验室研究表明，O3暴露会导致非人类的粘液细胞化生/增生灵长类动物，大鼠和小鼠。这包括我们小组的最新结果，证明了经典的近交暴露于0.8 ppm O3（每天4小时）的小鼠菌株9天在嗜酸性气道注射9天内粘液细胞增生。这是三种物种和人类的数据为旨在了解O3®非原子哮喘关联的潜在机制的研究。我们对机制问题采用遗传方法。我们调查了合作十字架的菌株（CC）小鼠种群，一个新的重组近交菌株，并确定了一种菌株CC002，即重复O3暴露后，暴露于嗜酸性粒细胞注射水平比任何其他菌株都要高得多迄今已测试。 O3暴露的CC002小鼠还会发展出粘液细胞化生和气道超级响应能力（AHR），另外两个哮喘的标志性表型。那就是CC002提供了一种新的遗传模型我们可以利用识别O3诱导的非原子哮喘关联的机制。在第一个目的，我们将检验以下假设：CC002对O3暴露的反应是由于通过警报模拟刺激2型先天淋巴样细胞（ILC2S）的警报器的上皮信号传导失调。我们将确定上皮衍生的先天细胞因子（IL-1A/B，IL-25，IL-33，TSLP）是否增加了O3暴露与无响应菌株相比，CC002小鼠，如果这些细胞因子的谈判会减弱O3诱导的气道注入和AHR。接下来，我们将确定在O3-中ILC2数量和激活是否增加暴露的CC002小鼠，如果ILC2耗尽或抑制ILC2衍生的细胞因子（IL-5，IL-13）会减轻 CC002小鼠中O3诱导的气道注射和AHR。全面表征分子对上皮，ILC2和其他白细胞中O3的反应，我们将执行单细胞RNA测序。使用这种创新和强大的方法，我们将量化基因表达和细胞组成的差异 +/- o3，无论是在应变内还是在菌株之间。我们还将通过测试检查CC002响应的特异性这种菌株是否表现出对其他暴露的反应，包括内毒素，香烟烟，和家尘螨过敏原。在第二个目标中，我们将确定对CC002敏感的遗传局部 O3。我们将执行定量性状位点映射以识别带有CC002等位基因的染色体区域与O3敏感性相关，然后进行生物信息学分析，以鉴定高优先级候选基因每个基因座。总的来说，我们的工作将揭示O3暴露与开发非原子哮喘表型并确定靶向治疗的潜在途径。