Genetic Control of Airway Epithelium Gene Expression in Childhood Asthmatics

儿童哮喘气道上皮基因表达的遗传控制

• 批准号: 9120408
• 负责人: Max A Seibold
• 金额: $ 61.67万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9120408
• 关键词: Affect; African American; Allelic Imbalance; Allergens; Asthma; Biological; Bronchoscopy; Cell Line; Child; Childhood; Childhood Asthma; Chromosome Mapping; Chronic Disease; Clinical; Code; Collection; Complex; DNA; Data; Development; Disease; Environmental Pollutants; Epithelial; Functional disorder; Gene Expression; Genes; Genetic; Genetic Predisposition to Disease; Genotype; Goals; Health; High Prevalence; Immune response; Interleukin-13; Mediating; Mesenchyme; Methods; Mexican; Molecular; Molecular Profiling; Mucous body substance; Nose; Obstruction; Pathogenesis; Pathway Analysis; Pathway interactions; Patients; Play; Population; Population Heterogeneity; Production; Proteins; Proxy; Puerto Rican; Quantitative Trait Loci; RNA Processing; Regulation; Regulatory Element; Risk; Role; Safety; Severities; Signal Transduction; Site; Susceptibility Gene; Technology; Testing; Tissues; Untranslated RNA; Variant; airway epithelium; airway inflammation; airway remodeling; asthmatic; bronchial epithelium; burden of illness; cost; differential expression; genome wide association study; genome-wide; human disease; microorganism; minority children; novel; risk variant; screening; targeted treatment; trait; transcriptome; transcriptome sequencing

 DESCRIPTION (provided by applicant): Asthma is the most common chronic disease among children. Results from gene mapping studies of complex human diseases including asthma have indicated that most associated variants are non-coding, rather than protein coding. Many of these non-coding risk variants likely reside in regulatory elements that affect gene expression. This suggests expression quantitative trait loci (eQTL) variants identified in a relevant tissue will present a variant pool enriched for asthma disease variants. The bronchial airway epithelium contributes to asthma pathogenesis by affecting airway immune responses, through the production of mucus that plugs airways, and by signaling to the airway mesenchyme which influences airway remodeling. Our goal is to identify airway epithelium eQTLs which confer risk of childhood asthma in diverse populations. The nasal airway epithelium is an easily accessible alternative to bronchial airway epithelium and the nasal airway transcriptome closely approximates the bronchial airway transcriptome. We hypothesize the functional underpinnings of many asthma genetic susceptibility loci involve cis-regulation of airway epithelium gene expression. We will test our hypothesis with three specific aims: Aim 1: Identify cis-regulated genes and their eQTLs within asthma GWAS loci in primary airway epithelium from Puerto Rican childhood asthmatics and controls. We will use targeted RNA-seq to determine total/allelic expression levels of all 159 genes localized to the 21 identified asthma GWAS loci in the nasal airway epithelium of 750 Puerto Rican asthmatics and controls. Cis-regulated genes will be determined by eQTL analyses using genome-wide genotype data and allelic imbalance analyses. Finemapping of eQTLs will be accomplished with complete variant data from targeted DNA-seq of all subjects. Aim 2: Identify novel asthma-related eQTLs by whole transcriptome eQTL analysis of primary airway epithelium from Puerto Rican childhood asthmatics and controls. Whole transcriptome expression on all 750 subjects will be used to identify genes that are both differentially expressed in asthma and are cis-regulated, as in Aim 1. DNA-seq will be used to generate complete variant data at the asthma-related, cis-regulated genes to perform eQTL finemapping. Aim 3: Characterize the lower airway, clinical, and biological significance of identified nasal asthma eQTLs in diverse populations of asthmatics. We will characterize the strongest nasal asthma eQTLs in the bronchial airway epithelium of 100 asthmatics. We will determine if the strongest finemapped airway eQTLs identified in Aims 1 and 2 predispose to asthma and related traits in 7200 Puerto Rican, Mexican, and African American children. Altered biologic mechanisms will be identified by pathway analysis of asthma-associated genes regulated by airway eQTLs.

 描述（由申请人提供）：哮喘是儿童中最常见的慢性疾病。对包括哮喘在内的复杂人类疾病的基因图谱研究结果表明，大多数相关变异是非编码的，而不是蛋白质编码的。许多非编码风险变异可能存在于影响基因表达的调控元件中。这表明在相关组织中鉴定出的表达数量性状位点（eQTL）变体将呈现出富含哮喘疾病变体的变体库。支气管气道上皮通过影响气道免疫反应、产生堵塞气道的粘液以及向气道间质发出信号影响气道重塑，从而促进哮喘发病机制。我们的目标是确定气道上皮 eQTL，这些 eQTL 会影响不同人群的儿童哮喘风险。鼻气道上皮是支气管气道上皮的容易获得的替代物，并且鼻气道转录组与支气管气道转录组非常接近。我们假设许多哮喘遗传易感性位点的功能基础涉及气道上皮基因表达的顺式调节。我们将通过三个具体目标来检验我们的假设： 目标 1：在波多黎各儿童哮喘患者和对照组的原发性气道上皮的哮喘 GWAS 位点内鉴定顺式调节基因及其 eQTL。我们将使用靶向 RNA 测序来确定 21 种已识别哮喘的所有 159 个基因的总/等位基因表达水平 750 名波多黎各哮喘患者和对照者鼻气道上皮中的 GWAS 位点。顺式调控基因将通过使用全基因组基因型数据和等位基因失衡分析的 eQTL 分析来确定。 eQTL 的精细定位将通过所有受试者的目标 DNA 测序的完整变异数据来完成。目标 2：通过对波多黎各儿童哮喘患者和对照组的原发气道上皮进行全转录组 eQTL 分析，识别新的哮喘相关 eQTL。所有 750 名受试者的全转录组表达将用于识别在哮喘中差异表达且受顺式调控的基因，如目标 1 所示。DNA-seq 将用于生成哮喘相关、顺式调控基因的完整变异数据，以执行 eQTL 精细定位。目标 3：表征不同哮喘人群中已识别的鼻哮喘 eQTL 的下呼吸道、临床和生物学意义。我们将描述 100 名哮喘患者的支气管气道上皮中最强的鼻哮喘 eQTL。我们将确定目标 1 和 2 中确定的最强精细气道 eQTL 是否容易导致 7200 名波多黎各、墨西哥和非裔美国儿童患哮喘及相关特征。通过对气道 eQTL 调节的哮喘相关基因进行通路分析，可以确定改变的生物学机制。