IL-13-induced SFRP1 requires STAT3 to regulate esophageal epithelial proliferation and Basal Zone Hyperplasia in EoE

IL-13诱导的SFRP1需要STAT3来调节EoE中的食管上皮增殖和基底区增生

基本信息

批准号：
10677306
负责人：
Sahiti Marella
金额：
$ 4.05万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2024-08-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10677306
关键词：
3-Dimensional Allergic Allergic Disease Attenuated Automobile Driving Backcrossings Binding Biological Biological Products Biopsy Biopsy Specimen CCL26 gene Cell Culture Techniques Cell Proliferation Cells ChIP-seq Chronic Clinical Computer Analysis DNA Binding Deglutition Disease Environment Eosinophilia Eosinophilic Esophagitis Epithelial Cell Proliferation Epithelial Cells Epithelium Equipment Esophageal Diseases Esophagus Extracellular Space FDA approved Food Foundations Functional disorder Gene Expression Genes Genetic Genetic Transcription Goals Histologic Human Hyperplasia Immune Impairment In Vitro Incidence Individual Inflammatory Inflammatory Response Interleukin-13 Interleukin-13 Overexpression Knowledge Link Mediating Medical Research Mentors Messenger RNA Modeling Molecular Mus Nucleic Acid Regulatory Sequences Ontology Outcome Pathogenesis Pathway interactions Patients Phenotype Prevalence Process Proliferating Proteins Quality of life Recombinants Regulation Research Research Personnel Research Training Role STAT3 gene STAT6 gene Scientist Signal Transduction Symptoms System Testing Time Training Up-Regulation WNT Signaling Pathway antagonist career cytokine differential expression eosinophil experience food allergen frizzled related protein-1 gastrointestinal symptom gene induction improved in vivo inhibitor insight mRNA Expression mast cell multidisciplinary novel novel therapeutics overexpression pharmacologic promoter small hairpin RNA therapeutic target three dimensional cell culture transcriptome transcriptome sequencing translational research program

项目摘要

ABSTRACT/PROJECT SUMMARY Eosinophilic Esophagitis (EoE) is a chronic allergic disease of the esophagus clinically characterized by symptoms including difficulty swallowing, food impaction, and decreased quality of life. Histopathological features of EoE include esophageal eosinophilia (EOS/HPF) and epithelial remodeling, including dilated intercellular spaces (DIS) and basal zone hyperplasia (BZH). BZH has been linked with a fibrostenotic phenotype and clinical outcomes such as esophageal stiffness and dysfunction, food impaction and stricture [1]. The underlying immune/inflammatory responses that drive EoE pathogenesis have been extensively studied; however, studies defining the molecular basis of esophageal epithelial proliferation and BZH in EoE are sparse. Herein, we identified a critical role Secreted Frizzled-Related Protein 1 (SFRP1) in driving esophageal epithelial proliferation in EoE. A major gap in our knowledge is the molecular basis of SFRP1 regulation in Interleukin-13 (IL-13)- induced esophageal epithelial proliferation and BZH in EoE. EoE disease pathogenesis is largely driven by the cytokine IL-13, which is upregulated in esophageal biopsies of EoE patients and is sufficient to alter gene expression in esophageal epithelial cells in vitro and in vivo. In preliminary studies we show that IL-13 induces a time-dependent increase in p-STAT3 (Y705 and S727) and p-STAT6 (Y641) in esophageal epithelial cells (EPC2-ALI) and in 3D primary esophageal cell cultures; however, IL-13-induced esophageal epithelial proliferation is STAT3-dependent. Differentially expressed genes (DEGs) derived from RNAseq analyses of esophageal biopsy samples from EoE individuals and IL-13-induced EPC2-ALI cells are enrichment for genes that contain putative STAT3 binding motifs and are enriched for genes involved in epithelial proliferation and are pro-survival pathways. We identified Secreted Frizzled Related Protein 1 (SFRP1), a soluble modulator of Wnt signaling and cellular proliferation as a common STAT3 putative target and DEG in esophageal epithelial cells. We will test the central hypothesis that IL-13-STAT3-dependent regulation of esophageal epithelial proliferation is mediated by SFRP1. The specific aims (SA) will SA1) Define the requirement of STAT3 in IL-13-induced SFRP1 expression in esophageal epithelial cells and SA2) Define the role of SFRP1 in IL-13-induced esophageal epithelial cell proliferation. My long-term career goal is to become an independent investigator in an academic setting. I will be immersed in an academically rich research training environment with access to world-class facilities, equipment, and basic / clinical / translational research programs. The proposed research will be supervised by a multidisciplinary team of expert scientists and clinicians, who have the experience and expertise to mentor me and assure successful completion of the proposed studies and provide an excellent foundation training to pursue a career in medical research.

摘要/项目摘要嗜酸性食管炎（EOE）是一种临床特征的食管的慢性过敏性疾病症状包括吞咽困难，食物施加和生活质量降低。组织病理学特征 EOE包括食管嗜酸性粒细胞（EOS/HPF）和上皮重塑，包括扩张的细胞间间的空间（DIS）和基础区域增生（BZH）。 BZH已与纤维状烯醇表型和临床联系在一起食管刚度和功能障碍，食物撞击和狭窄等结果[1]。基础驱动EOE发病机理的免疫/炎症反应已被广泛研究。但是，研究在EOE中定义食道上皮增殖和BZH的分子基础很少。这里，我们确定了与毛躁相关的蛋白1（SFRP1）在驱动食管上皮增殖中的关键作用在eoe。我们所知的一个主要差距是白介素13（IL-13）中SFRP1调节的分子基础 - 在EOE中诱导食管上皮增殖和BZH。 EOE疾病发病机理在很大程度上由细胞因子IL-13驱动，该因子在食管中被上调 EOE患者活检，足以改变食管上皮细胞中的基因表达体内。在初步研究中，我们表明IL-13诱导P-STAT3的时间依赖性增加（Y705和S727）食管上皮细胞（EPC2-ALI）和3D原发食道细胞培养物中的P-Stat6（Y641）；然而，IL-13诱导的食管上皮增殖是STAT3依赖性的。差异表达的基因（DEG）从EOE个体的食管活检样品和IL-13诱导的RNASEQ分析得出 EPC2-ALI细胞对含有推定的STAT3结合基序的基因富集，并富含基因参与上皮增殖，是临床途径。我们确定了分泌的毛躁相关蛋白 1（SFRP1），Wnt信号传导和细胞增殖的可溶调节器，作为常见的STAT3推定目标和食管上皮细胞的DEG。我们将测试IL-13-STAT3依赖性的中心假设食管上皮增殖的调节是由SFRP1介导的。具体目的（SA）将SA1）定义在IL-13诱导的食管上皮细胞中STAT3诱导的SFRP1表达的需求，SA2）定义 SFRP1在IL-13诱导的食管上皮细胞增殖中的作用。我的长期职业目标是成为学术环境中的独立研究员。我会的沉浸在一个学术丰富的研究培训环境中，可以使用世界一流的设施，设备，以及基本 /临床 /翻译研究计划。拟议的研究将由专家科学家和临床医生的多学科团队，他们具有指导我的经验和专业知识并确保成功完成拟议的研究，并提供出色的基础培训来追求医学研究的职业。