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

基本信息

批准号：
10321258
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
项目类别：
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/10321258
关键词：
3-Dimensional Accounting 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 Epithelial Cells 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 Supervision TLR2 gene Testing Tight Junctions Tissues Training Translational Research Up-Regulation Western Blotting Work allergic airway inflammation career chemokine chromatin immunoprecipitation chromatin modification 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

项目摘要

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.

项目概要该研究探讨了嗜酸粒细胞性食管炎上皮屏障功能障碍的机制 (EoE) 在其活动性疾病阶段，EoE 的特点是 Th2 型、富含嗜酸性粒细胞的炎症，并伴有高度的炎症反应。 IL-13、IL-4 和趋化因子的水平人们越来越认识到上皮功能障碍。由于对过敏原的敏感性，活动性 EoE 中的 EoE 可能与炎症成分发挥同样重要的作用此外，数据表明，对于许多患者来说，上皮功能障碍仍然存在。在非活动性 EoE 期间，治疗已清除了持续性上皮细胞的炎症浸润。然而，我们和其他人观察到了一种可重复的、失调的基因。活性 EoE 期间的表达模式与上皮结构蛋白的异常表达 EoE 患者的上皮细胞在生长环境中仍维持这种失调的基因表达模式。这表明表观遗传机制维持了这些基因的表达。在没有炎症的情况下 EoE 上皮的模式值得注意的是，对照上皮的体外 IL-13 处理。细胞复制 EoE 粘膜转录组的相似方面，表明该 Th2 细胞因子具有特定作用 EoE 的上皮屏障功能障碍与 IL-13 相关的上皮功能表观遗传改变有关。在几种类型的上皮细胞中都有显示，但食管中受影响的位点和功能意义是该提议的假设是 IL-13 诱导食管上皮的表观遗传变化。 EoE 患者，导致持续性上皮功能障碍这项工作将利用培训团队的帮助。拥有表观遗传学、转录组学和生物信息学方面的专业知识，具有与儿科合作的独特优势目标 1 将使用食管鳞状细胞的三维气液界面模型。上皮细胞以确定长期体外暴露于 IL-13 治疗是否会诱导表观遗传变化和屏障非 EoE 对照患者的原代上皮功能障碍本目的是确定 IL-13 是否存在。治疗与组蛋白标记的改变和已知影响基因的基因表达变化有关目标 2 将使用患有和未患有 EoE 的患者的活检组织来检查组蛋白的变化。该目的的目的是识别和定位一组食管上皮中的标记和基因表达。在活性和非活性 EoE 中持续表达失调的上皮基因（与对照相比）这些研究的结果将有助于了解食管上皮屏障功能障碍。和EoE疾病持续性，并为申请人首次提交R01提供依据。本提案中的研究策略和培训计划的目标是将候选人培养成独立的研究员领导一个强大的研究项目，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。