The Role of FOXM1 in Eosinophilic Esophagitis Pathogenesis

FOXM1 在嗜酸性食管炎发病机制中的作用

• 批准号: 10724896
• 负责人: Amanda Brooke Muir
• 金额: $ 17.8万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10724896
• 关键词: 3-Dimensional; Acids; Air; Allergic Disease; Allergic inflammation; Antigens; Award; Basal Cell Hyperplasia; Biological Assay; Body Weight decreased; CCL26 gene; Cell Culture Techniques; Cell Nucleus; Cells; Chemotactic Factors; Chemotaxis; Chronic Disease; Clinical; Cytoplasmic Granules; Data; Deglutition; Deglutition Disorders; Development; Disease; Disease remission; Down-Regulation; Endoscopy; Eosinophilic Esophagitis; Eosinophilic Infiltrate; Epithelial Cells; Epithelium; Esophagus; Evaluation; Event; FOXM1 gene; Fibrosis; Food; Future; Genes; Genetic; Goals; Health; Histologic; Homeostasis; Immune system; In Vitro; Incidence; Inflammation; Inflammatory; Interleukin-13; Interleukin-4; Intracellular Space; Invaded; Knowledge; Liquid substance; Modeling; Mucous Membrane; Mus; Organoids; Pathogenesis; Pathogenicity; Pathologic; Patients; Persons; Phosphorylation; Proliferating; Proteins; Quality of life; Refractory; Regulation; Research; Role; STAT6 Transcription Factor; STAT6 gene; Severity of illness; Techniques; Testing; Therapeutic; Tissues; United States; Up-Regulation; Vomiting; chromatin immunoprecipitation; cytokine; eosinophil; eosinophilic inflammation; forkhead protein; granulocyte; healing; human disease; in vivo; mouse model; novel; particle; pharmacologic; segregation; symptomatology; therapeutic target; transcription factor; transcriptome; treatment response; treatment strategy

Project Summary Eosinophilic esophagitis (EoE) is a rare, chronic disorder characterized by persistent allergic inflammation that leads to fibrosis and stricture which effects about 150,000 people in the United States. Clinical manifestations include weight loss, vomiting, dysphagia, and food impaction, all of which dramatically impact patients' quality of life. Histologically, it is characterized by mucosal eosinophilic infiltration and esophageal epithelial remodeling events, specifically basal cell hyperplasia (BCH) and dilated intracellular spaces (DIS). These epithelial changes disrupt the mucosal barrier which normally provides protection from acid and food particles during normal swallows. In our preliminary data we have found that Forkhead box (FOX) M1 expression is increased in the esophagus of EoE patients. Further, FOXM1 is induced in the epithelium after stimulation with IL-13, the major effector cytokine in EoE. We have found that inhibition of FOXM1 in esophageal epithelial cell culture reduces epithelial perturbations in the setting of cytokine stimulation and reduces chemoattractant secretion. Based on these findings, we hypothesize that FOXM1 disrupts epithelial homeostasis and contributes to eosinophil chemotaxis in EoE We propose 2 aims in this application. The first aim will determine the mechanism by which FOXM1 disrupts epithelial homeostasis in the esophageal epithelium using 3D epithelial modeling techniques that are well established in the Muir Lab: organoid culture and air-liquid interface culture. We will induce EoE in mice and evaluate the effect of FOXM1 inhibition on disease severity and epithelial damage. We will compliment this with chromatin immunoprecipitation to identify differentially regulated targets of FOXM1. In the second aim we will define the role of FOXM1 in regulation of chemotaxis in the EoE epithelium. We have found that pharmacological inhibition of FOXM1 leads to downregulation of both total and phosphorylated STAT6 protein, as well as CCL26 (the gene encoding Eotaxin-3, the key chemoattractant for eosinophils in EoE). We will test whether FOXM1 inhibition interferes with eosinophil chemotaxis in vitro and in vivo. Thisinnovative and hypothesis-driven study is backed by strong preliminary data generated by the PI. The PI is uniquely poised to accomplish these aims with her previous track record in investigating mechanisms of epithelial inflammation in EoE using 3D epithelial culture and murine models.

项目摘要 嗜酸性粒细胞性食管炎是一种罕见的慢性疾病， 导致纤维化和狭窄的炎症，在美国影响了大约15万人。临床 临床表现包括体重减轻、呕吐、吞咽困难和食物嵌塞，所有这些都会严重影响 患者的生活质量。组织学上，其特征是粘膜嗜酸性粒细胞浸润和食管癌。 上皮重塑事件，特别是基底细胞增生（BCH）和扩张的细胞内空间（DIS）。 这些上皮的变化破坏了粘膜屏障，而粘膜屏障通常提供对酸和食物的保护 正常吞咽时的颗粒物。在我们的初步数据中，我们发现叉头盒（FOX）M1表达 在EoE患者的食管中增加。此外，FOXM 1在上皮细胞中被诱导， IL-13是EoE中的主要效应细胞因子。我们发现抑制食管上皮细胞FOXM 1的表达， 培养减少了细胞因子刺激环境中的上皮扰动， 分泌物。基于这些发现，我们假设FOXM 1破坏了上皮细胞的稳态， 嗜酸性粒细胞趋化性 我们在本申请中提出了两个目标。第一个目标将确定FOXM 1 使用3D上皮建模技术破坏食管上皮的上皮稳态， 在Muir实验室中已建立良好的：类器官培养和气液界面培养。我们将在小鼠中诱导EoE， 评估FOXM 1抑制对疾病严重程度和上皮损伤的影响。我们会用 染色质免疫沉淀以鉴定FOXM 1的差异调节靶点。在第二个目标中我们将 定义FOXM 1在EoE上皮细胞趋化性调节中的作用。我们发现药理学上 FOXM 1的抑制导致总的和磷酸化的STAT 6蛋白以及CCL 26的下调 (the编码Eotaxin-3的基因，EoE中嗜酸性粒细胞的关键化学引诱物）。我们将测试FOXM 1是否 抑制在体外和体内干扰嗜酸性粒细胞趋化性。 这项创新和假设驱动的研究得到了PI生成的强有力的初步数据的支持。 PI凭借其先前在调查机制方面的记录， 使用3D上皮培养和小鼠模型在EoE中观察上皮炎症。