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/项目摘要嗜酸性粒细胞性食管炎（EoE）是一种慢性食管过敏性疾病，临床特征为症状包括吞咽困难、食物嵌塞和生活质量下降。组织病理学特征 EoE 的影响包括食管嗜酸性粒细胞增多 (EOS/HPF) 和上皮重塑，包括细胞间质扩张间隙（DIS）和基底区增生（BZH）。 BZH 与纤维狭窄表型和临床表现有关食管僵硬和功能障碍、食物嵌塞和狭窄等结果[1]。底层的驱动 EoE 发病机制的免疫/炎症反应已得到广泛研究；然而，研究定义 EoE 中食管上皮增殖和 BZH 的分子基础很少。在此，我们确定了分泌性卷曲相关蛋白 1 (SFRP1) 在驱动食管上皮增殖中的关键作用在EoE中。我们知识中的一个主要空白是白细胞介素 13 (IL-13) 中 SFRP1 调节的分子基础 - EoE 中诱导食管上皮增殖和 BZH。 EoE 疾病的发病机制很大程度上是由细胞因子 IL-13 驱动的，该细胞因子在食管中表达上调 EoE 患者的活检足以改变体外和体内食管上皮细胞的基因表达体内。在初步研究中，我们表明 IL-13 会诱导 p-STAT3（Y705 和 S727）呈时间依赖性增加食管上皮细胞 (EPC2-ALI) 和 3D 原代食管细胞培养物中的 p-STAT6 (Y641)；然而，IL-13 诱导的食管上皮增殖是 STAT3 依赖性的。差异表达基因（DEG）源自 EoE 个体和 IL-13 诱导的食管活检样本的 RNAseq 分析 EPC2-ALI 细胞富集含有推定 STAT3 结合基序的基因参与上皮增殖并且是促生存途径。我们鉴定出分泌性卷曲相关蛋白 1 (SFRP1)，Wnt 信号传导和细胞增殖的可溶性调节剂，作为常见 STAT3 假定靶点和食管上皮细胞中的DEG。我们将检验 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。具体目标 (SA) 将 SA1) 定义 IL-13 诱导食管上皮细胞中 SFRP1 表达对 STAT3 的要求和 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者。我会的沉浸在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，以及基础/临床/转化研究项目。拟议的研究将由由专家科学家和临床医生组成的多学科团队，他们拥有丰富的经验和专业知识来指导我并确保成功完成拟议的研究，并提供良好的基础培训以追求从事医学研究的职业。