Epigenetic regulation of esophageal epithelial barrier function in eosinophilic esophagitis

嗜酸性粒细胞性食管炎食管上皮屏障功能的表观遗传调控

• 批准号: 10531869
• 负责人: Melanie A Ruffner
• 金额: $ 18.53万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10531869
• 关键词: 3-Dimensional; Affect; Age; Air; Allergens; Allergic; Automobile Driving; Basal Cell Hyperplasia; Behavior; Bioinformatics; Biopsy; Cell division; Cell physiology; Cells; ChIP-seq; Chemotactic Factors; Childhood; Chronic; DNA Sequence; Data; Disease; Down-Regulation; Drug Targeting; Environment; Eosinophilic Esophagitis; Eosinophilic Infiltrate; Epigenetic Process; Epithelial Cells; Epithelium; Esophageal mucous membrane; Esophagus; Exposure to; Flow Cytometry; Food Hypersensitivity; Foundations; Functional disorder; Future; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Proteins; Genes; Genetic Transcription; Genome; Goals; Growth; Heritability; Heterogeneity; Histologic; Histones; Human; Immunofluorescence Immunologic; Impairment; In Vitro; Inflammation; Inflammatory; Inflammatory Infiltrate; Interleukin-13; Interleukin-4; Intracellular Space; Liquid substance; Mediating; Medical; Mentors; Microbe; Modeling; Modification; Morbidity - disease rate; Mucositis; Mucous Membrane; Outcome Study; Pathogenesis; Pathway interactions; Patients; Persons; Phase; Phenotype; Play; Population; Positioning Attribute; Process; Proteins; Quantitative Reverse Transcriptase PCR; Regulatory Pathway; Reproducibility; Research; Research Personnel; Resolution; Risk; Role; Signal Pathway; Signal Transduction; Squamous Differentiation; Squamous Epithelium; Structural Protein; Structure; TLR2 gene; Tight Junctions; Tissues; Training; Translational Research; Up-Regulation; Western Blotting; Work; allergic airway inflammation; barrier to testing; career; chemokine; chromatin immunoprecipitation; chromatin modification; comparison control; cytokine; dietary; eosinophil; eosinophilic inflammation; epigenetic regulation; experience; genomic locus; histone modification; improved; insight; microorganism antigen; novel; programs; protein expression; single-cell RNA sequencing; skills; transcription factor; transcriptome; transcriptomics

PROJECT SUMMARY The proposed research explores the mechanisms of epithelial barrier dysfunction in eosinophilic esophagitis (EoE). During its active disease phase, EoE is characterized by Th2-type, eosinophil-rich inflammation with high levels of IL-13, IL-4 and chemoattractant chemokines. There is a growing appreciation that epithelial dysfunction in active EoE may play an equally important role as the inflammatory component due to sensitization to allergens and exposure to microbes. Additionally, data suggests that for many patients, epithelial dysfunction persists during inactive EoE when therapy has cleared the inflammatory infiltrate. The mechanism of persistent epithelial dysfunction is poorly understood. However, we and others have observed a reproducible, dysregulated gene expression pattern during active EoE with aberrant expression of epithelial structural proteins. We show that epithelial cells from EoE patients maintain aspects of this dysregulated gene expression pattern when grown in a neutral culture environment. This suggests that epigenetic mechanisms maintain these gene expression patterns in EoE epithelium in the absence of inflammation. Notably, in vitro IL-13 treatment of control epithelial cells replicates similar aspects of the EoE mucosal transcriptome, suggesting a specific role for this Th2 cytokine in the epithelial barrier dysfunction of EoE. IL-13-associated epigenetic alteration of epithelial function has been shown in several types of epithelium but the affected loci and functional implications in the esophagus are unclear. The hypothesis of this proposal is that IL-13 induces epigenetic changes in the esophageal epithelium of patients with EoE, leading to persistent epithelial dysfunction. The work will leverage access to a training team with expertise in epigenetics, transcriptomics and bioinformatics is uniquely situated to work with a pediatric population of EoE patients. Aim 1 will use a three dimensional air-liquid interface model of esophageal squamous epithelium to determine if chronic in vitro exposure to IL-13 treatment induces epigenetic changes and barrier dysfunction in primary epithelium from non-EoE control patients. The goal of this aim is to identify if IL-13 treatment is associated with alterations in histone marks and gene expression changes in genes known to affect barrier function. Aim 2 will use biopsy tissue from patients with and without EoE to examine changes in histone marks and gene expression in the esophageal epithelium. The goal of this aim is to identify and localize a set of epithelial genes with persistently dysregulated expression in active and inactive EoE (when compared to control patients). The outcome of these studies will inform the understanding of esophageal epithelial barrier dysfunction and EoE disease persistence, and provide the basis for the applicant’s first R01 submission. The overarching goal of the research strategy and training plan in this proposal is to develop the candidate into an independent investigator leading a robust research program that utilizes research and analysis skills in epigenetics, transcriptomics and bioinformatics in the field of non-IgE mediated food allergies.

项目摘要 拟议的研究探讨了嗜酸性静脉炎中上皮屏障功能障碍的机制 （eoe）。在其活性疾病阶段，EOE的特征是Th2型，富含嗜酸性粒细胞的注射 IL-13，IL-4和趋化剂趋化因子的水平。越来越多的上皮功能障碍 在主动EOE中，由于对过敏原的敏感性，可能与炎症成分一样重要 并暴露于微生物。此外，数据表明，对于许多患者，上皮功能障碍持续存在 在不活跃的EOE期间，当治疗已清除炎症性浸润。持续上皮的机制 功能障碍知之甚少。但是，我们和其他人观察到了可再现的，失调的基因 活性EOE期间的表达模式与上皮结构蛋白异常表达。我们表明 来自EOE患者的上皮细胞在生长中生长时保持这种失调的基因表达模式的各个方面 中立的文化环境。这表明表观遗传机制保持这些基因表达 在没有炎症的情况下，EOE上皮的模式。值得注意的是，在体外IL-13对照上皮的处理 细胞复制EOE粘膜转录组的相似方面，这表明该Th2细胞因子的特定作用 在EOE的上皮屏障功能障碍中。 IL-13相关上皮功能的上皮改变已经 在几种类型的上皮中显示，但食管中受影响的局部和功能影响是 该提议的假设是IL-13诱导食管上皮的表观遗传变化 EOE的患者，导致持续的上皮功能障碍。这项工作将利用培训团队的访问权限 具有表观遗传学的专业知识，转录组学和生物信息学是独特的，可以与儿科一起工作 EOE患者的人口。 AIM 1将使用食管鳞状的三维空气接口模型 上皮以确定慢性体外暴露于IL-13治疗是否诱导表观遗传变化和障碍 非EOE对照患者的原发性上皮功能障碍。该目标的目的是确定IL-13是否 治疗与已知影响的基因的组蛋白标记变化和基因表达变化有关 屏障功能。 AIM 2将使用或不带EOE患者的活检组织检查组蛋白的变化 食管上皮中的标记和基因表达。此目的的目的是识别和本地化一组 在活性和非活性EOE中表达持续失调的上皮基因（与对照相比 患者）。这些研究的结果将告知对食道上皮屏障功能障碍的理解 和EOE疾病的持久性，并为申请人的第一个R01提交提供了基础。总体 该提案中研究策略和培训计划的目标是将候选人发展成独立 研究人员领导了一项强大的研究计划，该计划利用表观遗传学的研究和分析技能， 非IGE介导的食物过敏领域的转录组学和生物信息学。