Transcriptional Responses to Wildfire Pollution in Airway Epithelial Cells Identify Genetic Risk Factors and Mechanisms of Asthma Exacerbations

气道上皮细胞对野火污染的转录反应确定了哮喘恶化的遗传风险因素和机制

• 批准号: 10573003
• 负责人: Arnav Gupta
• 金额: $ 17.34万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-17 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10573003
• 关键词: Address; Air; Air Pollutants; Air Pollution; Alleles; Aryl Hydrocarbon Receptor; Asthma; Binding; Binding Sites; Bioinformatics; Biological Assay; Cell Culture Techniques; Cell Line; Cell model; Chromatin; Clinical; Clinical Data; Clustered Regularly Interspaced Short Palindromic Repeats; Consensus; Dexamethasone; Disease; Disease model; Epithelial Cells; Family; Gene Expression; Genes; Genetic Polymorphism; Genetic Transcription; Genomic DNA; Genomics; Glucocorticoid Receptor; Grant; Health; Housing; Human; IL8 gene; Individual; Inflammation; Interleukin-8; Laboratories; Liquid substance; Luciferases; Mediating; Mediator; Mentors; Methodology; Methods; Minor; Modeling; Morbidity - disease rate; NF-kappa B; Particulate; Patients; Physiological; Plasmids; Pollution; Population Study; Predisposition; Public Health; Recommendation; Regulation; Regulator Genes; Reporter; Risk Estimate; Role; Series; Severities; Single Nucleotide Polymorphism; Smoke; Susceptibility Gene; Symptoms; Techniques; Testing; Toxic effect; Variant; Wildfire; airway epithelium; associated symptom; asthma exacerbation; bioinformatics tool; climate change; experience; genetic risk factor; genetic variant; genome editing; high risk; human tissue; mortality; next generation sequencing; novel; particle; particle exposure; pharmacologic; pollutant; receptor expression; response; therapeutic target; tissue culture; tool; transcription factor; wood smoke

Project Summary/Abstract Increased levels of air pollution from wildfires are directly associated with asthma exacerbations but the mechanisms mediating this association are not understood. Furthermore, the severity of asthma symptoms that result from wildfire exposure vary considerably by individual. We seek to identify Transcriptional Regulatory Elements (TREs) that connect wildfire particulate exposures with genetic variants associated with asthma to identify susceptibility features and mechanisms that govern asthma exacerbations. We will employ wood smoke particle (WSP) exposure in primary airway epithelial cells cultured at air- liquid interface to model the interaction between the respiratory epithelium and fine particulates released by wildfires. We will assay chromatin accessibility and nascent transcription using next- generation sequencing techniques to identify TREs regulated by WSP. Then, using a permutation- based bioinformatic approach, we will intersect the genomic coordinates of these TREs with genomic susceptibility loci for asthma. We will then characterize the transcriptional function of single nucleotide polymorphisms (SNPs) that result from this intersection. We will test TRE function in the context of WSP exposure with and without the SNPs in plasmid-derived luciferase reporters and in genomic DNA, using the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) – Cas9 tool to introduce the variant alleles. Finally, to assay the physiologic significance of these TRE-SNP overlaps, we will test the role of the rs258760 SNP, which inhabits a TRE under regulation by WSP and controls expression of the glucocorticoid receptor, in mediating Interleukin-8 secretion by exposing airway epithelial cells from donors with the major and minor alleles to WSP and dexamethasone. This project will identify and characterize genomic features that connect air pollution with asthma exacerbations. These loci and the genes they regulate will serve as candidates for pharmacologic therapies to mitigate asthma symptoms associated with wildfire air pollution. Furthermore, this project develops a method to integrate multiple models of disease in a genomic context. This strategy may be used to propose novel treatments for asthma.

项目摘要/摘要 野火的空气污染水平增加与哮喘病变直接相关，但 介导这种关联的机制不理解。此外，哮喘的严重程度 野火暴露导致的症状因个人而谨慎而异。我们试图确定 转录调节元件（TRE）将特定暴露与遗传联系起来 与哮喘相关的变体，以识别控制哮喘的敏感性特征和机制 恶化。 我们将在空气中培养的原发气道上皮细胞中采用木烟颗粒（WSP）暴露 液态界面，以建模呼吸上皮和细节之间的相互作用 由野火发布。我们将使用下一步 生成测序技术以识别由WSP调节的TRE。然后，使用置换 - 基于生物信息学方法，我们将与基因组相交这些TRE的基因组坐标 哮喘的易感性位置。然后，我们将表征单个核苷酸的转录功能 由该交叉路口产生的多态性（SNP）。我们将在 WSP在质粒衍生的荧光素酶报告中和基因组中有和没有SNP的WSP暴露 DNA，使用簇的定期间隔短的短文重复序列（CRISPR） - Cas9工具 介绍变体等位基因。最后，为了确保这些TRE-SNP重叠的生理意义， 我们将测试RS258760 SNP的作用，该RS258760 SNP会影响WSP和控制的调节的TRE 糖皮质激素受体的表达，通过暴露气道介导白介素-8分泌 来自主要和小等位基因WSP和地塞米松的供体的上皮细胞。 该项目将识别并表征将空气污染与哮喘联系起来的基因组特征 恶化。这些基因座及其调节的基因将用作药理学的候选者 减轻与野火空气污染相关的哮喘症状的疗法。此外，这个 项目开发了一种在基因组环境中整合多种疾病模型的方法。这 策略可用于提出针对哮喘的新型治疗方法。