The Role of FOXM1 in Eosinophilic Esophagitis Pathogenesis

项目摘要 嗜酸性食管炎(EoE)是一种罕见的慢性疾病，以持续性过敏为特征。 导致纤维化和狭窄的炎症，在美国大约有15万人受到影响。临床 表现包括体重减轻、呕吐、吞咽困难和食物嵌塞，所有这些都会对 患者的生活质量。组织学上以粘膜嗜酸性粒细胞浸润和食道为特征。 上皮重塑事件，特别是基底细胞增生(BCH)和细胞内间隙扩张(DIS)。 这些上皮细胞的变化破坏了正常情况下提供酸和食物保护的粘膜屏障。 正常吞咽过程中的颗粒。在我们的初步数据中，我们已经发现Forkhead Box(Fox)M1表达 在EoE患者的食道中增加。此外，FOXM1在刺激后的上皮细胞中被诱导。 IL-13是EoE中的主要效应细胞因子。我们发现FOXM1在食道上皮细胞中的抑制作用 培养减少细胞因子刺激环境中的上皮扰动和减少趋化物质 分泌物。基于这些发现，我们假设FOXM1破坏了上皮细胞的动态平衡，并对 嗜酸性粒细胞在EoE中的趋化作用 在本申请中，我们提出了两个目标。第一个目标将决定FOXM1通过什么机制 使用3D上皮建模技术破坏食道上皮的上皮内稳态 在缪尔实验室很好地建立了：有机物培养和气液界面培养。我们将在小鼠体内诱导EoE并 评估FOXM1抑制对疾病严重程度和上皮损伤的影响。我们将用以下几个方面加以补充 染色质免疫沉淀鉴定FOXM1的差异调控靶点。在第二个目标中，我们将 明确FOXM1在EoE上皮趋化性调节中的作用。我们发现从药理上讲 抑制FOXM1导致总的和磷酸化的STAT6蛋白以及CCL26的下调 (编码嗜酸性粒细胞趋化因子-3的基因，嗜酸性粒细胞在嗜酸性粒细胞中的关键趋化因子)。我们将测试FOXM1是否 抑制在体外和体内干扰嗜酸性粒细胞的趋化作用。 这项创新和假设驱动的研究得到了PI产生的强大初步数据的支持。 私家侦探凭借她之前在调查机制方面的记录，为实现这些目标做好了准备 使用3D上皮细胞培养和小鼠模型研究EoE中的上皮炎症。

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.