The Role of FOXM1 in Eosinophilic Esophagitis Pathogenesis

项目概要 嗜酸性粒细胞性食管炎 (EoE) 是一种罕见的慢性疾病，其特征是持续过敏 导致纤维化和狭窄的炎症影响了美国约 150,000 人。临床 症状包括体重减轻、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，其特点是粘膜嗜酸性粒细胞浸润和食管 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）。 这些上皮变化破坏了通常提供免受酸和食物侵害的粘膜屏障 正常吞咽时的颗粒。在我们的初步数据中，我们发现 Forkhead box (FOX) M1 表达 EoE 患者的食道中增加。此外，FOXM1 在刺激后在上皮细胞中被诱导。 IL-13，EoE 中的主要效应细胞因子。我们发现食管上皮细胞中FOXM1的抑制 培养减少细胞因子刺激环境下的上皮扰动并减少化学引诱剂 分泌。基于这些发现，我们假设 FOXM1 破坏上皮稳态并有助于 EoE 中的嗜酸性粒细胞趋化作用 我们在此应用中提出 2 个目标。第一个目标将确定 FOXM1 的机制 使用 3D 上皮建模技术破坏食管上皮的上皮稳态 缪尔实验室已确立：类器官培养和气液界面培养。我们将在小鼠中诱导 EoE 评估 FOXM1 抑制对疾病严重程度和上皮损伤的影响。我们将称赞这一点 染色质免疫沉淀来识别 FOXM1 的差异调节靶点。在第二个目标中，我们将 定义 FOXM1 在 EoE 上皮趋化性调节中的作用。我们发现，药理 抑制 FOXM1 会导致 STAT6 总蛋白和磷酸化 STAT6 蛋白以及 CCL26 下调 （编码 Eotaxin-3 的基因，EoE 中嗜酸性粒细胞的关键化学引诱剂）。我们将测试FOXM1是否 抑制作用会干扰体外和体内嗜酸性粒细胞的趋化性。 这项创新且以假设为驱动的研究得到了 PI 生成的强有力的初步数据的支持。 PI 凭借她之前在调查机制方面的记录，具备独特的能力来实现这些目标 使用 3D 上皮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.