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

• 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
• 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

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/项目总结 嗜酸性食管炎（EoE）是一种慢性过敏性食管疾病，其临床特征为： 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包括食管嗜酸性粒细胞增多（EOS/HPF）和上皮重塑，包括扩张的细胞间质 基底区增生（BZH）。BZH与纤维狭窄表型和临床 结果如食管僵硬和功能障碍、食物嵌塞和狭窄[1]。底层 已经广泛研究了驱动EoE发病机制免疫/炎症反应;然而， 明确EoE中食管上皮增殖和BZH的分子基础的文献很少。在此我们 确定了分泌卷曲相关蛋白1（SFRP 1）在驱动食管上皮增殖中的关键作用 在EoE。我们知识的一个主要空白是SFRP 1调节白细胞介素-13（IL-13）的分子基础。 诱导EoE中食管上皮增殖和BZH。 EoE疾病的发病机制在很大程度上是由细胞因子IL-13驱动的，IL-13在食管癌中上调， EoE患者的活组织检查，并足以改变体外食管上皮细胞的基因表达， vivo.在初步研究中，我们表明IL-13诱导p-STAT 3（Y 705和S727）的时间依赖性增加。 和食管上皮细胞（EPC 2-ALI）和3D原代食管细胞培养物中的p-STAT 6（Y 641）; 然而，IL-13诱导的食管上皮增殖是STAT 3依赖性的。差异表达基因 来自EoE个体的食管活检样品的RNAseq分析和来自IL-13诱导的EoE个体的食管活检样品的RNAseq分析的DEG（DEG）。 EPC 2-ALI细胞富集了含有推定的STAT 3结合基序的基因，并且富集了 参与上皮细胞增殖，是促生存途径。我们鉴定了分泌卷曲相关蛋白 1（SFRP 1），Wnt信号和细胞增殖的可溶性调节剂，作为常见的STAT 3推定靶点 和DEG。我们将检验IL-13-STAT 3依赖性细胞因子的中心假设， 食管上皮增殖的调节由SFRP 1介导。具体目标（SA）将SA 1）定义 STAT 3在IL-13诱导的食管上皮细胞SFRP 1表达中的需求和SA 2）定义 SFRP 1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调查员。我将 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， 基础/临床/转化研究项目。拟议的研究将由一名 我有一个多学科的专家科学家和临床医生团队，他们有经验和专业知识来指导我 并确保顺利完成拟议的研究，并提供一个良好的基础培训，以追求 从事医学研究