SLC9A3 regulation of esophageal dilated intercellular spaces in EoE Subtypes

SLC9A3 对 EoE 亚型食管扩张细胞间隙的调节

• 批准号: 9919496
• 负责人: SIMON Patrick HOGAN
• 金额: $ 62.45万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-22 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9919496
• 关键词: Acids; Adult; Allergic; Allergic inflammation; Animals; Aryl Hydrocarbon Receptor; Automobile Driving; Biopsy; Biopsy Specimen; CCL26 gene; Calpain; Cells; Child; Clinical; Communities; Coupled; Data Set; Deglutition Disorders; Development; Diet; Disease; Environmental Risk Factor; Eosinophilia; Eosinophilic Esophagitis; Epithelial; Epithelial Cells; Epithelium; Esophageal Diseases; Esophageal Squamous Cell; Esophagus; Extracellular Space; Family member; Food; Functional disorder; Gastroesophageal reflux disease; Gene Expression; Genes; Genetic; Genetic Transcription; Histologic; Human; Hyperplasia; Immune; Immunity; Individual; Inflammatory Response; Interleukin-13; Intervention; Kininogenase; Mediating; Medical; Molecular; Ontology; Organoids; Outcome Study; Pathway interactions; Patients; Pharmacology; Predisposition; Prevalence; Process; Proteins; Proton Pump Inhibitors; Protons; RNA; RNA Interference; Receptor Signaling; Regulation; Regulator Genes; Resistance; Serine Protease; Severity of illness; Signal Transduction; Sodium-Hydrogen Antiporter; Squamous Epithelium; Stratum Basale; System; Techniques; Technology; Testing; Therapeutic; Transcript; Transcriptional Regulation; Transplantation; aryl hydrocarbon receptor ligand; base; chronic inflammatory disease; cohort; cytokine; desmoglein 1; differential expression; eosinophil; eosinophilic inflammation; food antigen; gastrointestinal symptom; genetic profiling; humanized mouse; insight; intraepithelial; mRNA Expression; patient subsets; protein expression; transcription factor; transcriptome; transcriptome sequencing

Project Summary Eosinophilic esophagitis (EoE) is an increasingly prevalent chronic inflammatory disease of the esophagus, mediated by dietary food antigens and clinically characterized by upper gastrointestinal (GI) symptoms including dysphagia and food impaction. Recently, a confounding esophageal disorder, termed proton-pump inhibitor (PPI)–responsive esophageal eosinophilia (PPI-REE), has emerged; PPI-REE is indistinguishable from EoE by clinical, endoscopic or histologic features or by gene profiles. The current clinical conundrums are whether PPI- REE represents a GERD-related phenomenon, a subtype of EoE or a completely new entity and why PPI-REE and EoE respond differently to PPI. RNA sequencing (RNA-Seq) analyses of esophageal biopsy samples from patients with active EoE disease revealed dysregulation of gene networks associated with regulating intracellular [pH]i and acid protection and that the most upregulated transmembrane transporter activity gene was SLC9A3, which encodes for the sodium-hydrogen exchanger family member 3 (NHE3). Recently, we have demonstrated 1) increased expression of SLC9A3 within the basal layer of ESSE biopsies from patients with EoE and that expression positively correlated with disease severity (eosinophils/HPF) and DIS; 2) IL-13 induced SLC9A3 expression and function in ESSE cells and that SLC9A3 activity positively correlating with DIS formation and 3) SLC9A3-mediated Na+-dependent proton secretion is the primary intracellular acid protective mechanism within IL-13–stimulated ESSE cells and blockade of this pathway abrogated DIS formation45. In new preliminary studies we have made several transformative observations: 1) IL-13 induced expression of the transcription factor aryl hydrocarbon receptor (AhR) and AhR-responsive genes in ESSE cells and EoE biopsies; 2) stimulating ESSE cells with AhR ligands, suppressed AhR-responsive gene expression including SLC9A3 and 3) a divergent effect of PPI therapy on SLC9A3 expression in ESSE biopsy samples from individuals with EoE and PPI-REE, suggesting an opposing impact of PPI on SLC9A3 transcriptional regulation between EoE and PPI-REE. Collectively, these observations underlie our central hypothesis that SLC9A3 activity promotes DIS formation in EoE subtypes and that this pathway is divergently responsive to PPI therapy via AhR-dependent signaling. The specific Aims outlined in this proposal will 1) Aim 1. Determine the relationship between SLC9A3 expression and function, disease severity and DIS formation in EE subtypes; 2) Define the requirement of SLC9A3 in ESSE DIS formation and 3) Define the involvement of PPI-induced AhR signaling in Type-2 cytokine-induced SLC9A3 expression and function in ESSE cells. With respect to the expected outcomes, the studies proposed in Aim I are expected to establish the contribution of SLC9A3 to histopathologic features of EoE and PPI-REE and responsiveness of this pathway to PPI trial; Aim II are expected to determine the necessity of SLC9A3 to ESSE acid transport and DIS and Aim III is expected to determine the interaction between IL-13– and PPI-induced AhR signaling in SLC9A3 expression and function in ESSE cells and DIS formation. Successfully completing the proposed studies will provide a new and substantive departure from our current understanding of the underlying molecular mechanisms underpinning the development of PPI-REE and EoE and provide an explanation for their differential responsiveness to PPI therapy, thereby directing the development of new and pre-existing therapeutics for treating esophageal eosinophilia–related disorders.

项目摘要 嗜酸性食管炎（EOE）是一种日益普遍的Eosphagus的慢性炎性疾病， 由饮食中的食物抗原介导，临床特征是上胃肠道（GI）症状，包括 吞咽困难和食物影响。最近，一种混杂的食管疾病，称为质子 - 泵抑制剂 （PPI） - 反应性食管嗜酸性粒细胞（PPI-REE）已出现； PPI-ree与EOE无法区分 临床，内窥镜或组织学特征或基因谱。当前的临床难题是PPI- REE代表与GERD相关的现象，EOE的亚型或一个全新的实体，以及为什么PPI-ree EOE对PPI的反应不同。 RNA测序（RNA-SEQ）分析食管活检样品的分析 活跃EOE疾病的患者显示，与细胞内调节性相关的基因网络失调 [pH] i和酸保护，并且最新更新的转运蛋白活性基因是slc9a3， 该编码为氢交换器家族成员3（NHE3）编码。最近，我们已经证明了 1）在EOE患者的ESS活检基本层中SLC9A3的表达增加，并且 表达与疾病严重程度（嗜酸性粒细胞/HPF）和DIS呈正相关； 2）IL-13诱导SLC9A3 ESSE细胞中的表达和功能，SLC9A3活性与DIS形成正相关，3） SLC9A3介导的Na+依赖性质子分泌是在细胞内酸的主要保护机制 IL-13刺激的ESS细胞和该途径的阻塞杂技45。在新的初步研究中 我们进行了几个变革性观察：1）IL-13诱导的转录因子芳基的表达 ESSE细胞和EOE活检中的碳氢化合物受体（AHR）和AHR反应基因； 2）刺激Esse 具有AHR配体的细胞，抑制AHR响应基因表达，包括SLC9A3和3） 来自EOE和PPIRE的个体ESS活检样品中SLC9A3表达的PPI治疗 表明PPI对EOE和PPI Ree之间的SLC9A3转录调控产生了相反的影响。 总的来说，这些观察结果是我们的核心假设，即SLC9A3活性促进了DIS的形成 在EOE亚型中，该途径通过AHR依赖性信号传导对PPI治疗有多样化。 本提案中概述的具体目的将为1）目标1。确定SLC9A3表达式之间的关系 EE亚型中的功能，疾病的严重程度和疾病的形成； 2）在ESSE中定义SLC9A3的要求 DIS形成和3）定义PPI诱导的AHR信号在2型细胞因子诱导的SLC9A3中的参与 ESSE细胞中的表达和功能。关于预期的结果，目的I提出的研究 期望建立SLC9A3对EOE和PPI-REE的组织病理学特征的贡献 对PPI试验途径的响应能力； AIM II有望确定SLC9A3的必要条件 酸的运输和DIS和AIM III有望确定IL-13-和PPI诱导的相互作用 SLC9A3中的AHR信号传导在ESSE细胞和DIS形成中的表达和功能。成功完成 拟议的研究将与我们当前对基础的理解相偏离新的实质性。 PPIRE和EOE的发展的分子机制，并为其提供解释 对PPI疗法的反应性差异，从而指导新的和现有的 治疗食管嗜酸性粒细胞相关疾病的治疗剂。