IL-13-induced SFRP1 requires STAT3 to regulate esophageal epithelial proliferation and Basal Zone Hyperplasia in EoE

摘要/项目摘要 嗜酸性食管炎(EoE)是一种慢性食道过敏性疾病，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包括食管嗜酸性粒细胞增多症(EOS/HPF)和上皮重构，包括细胞间扩张 间隙(DIS)和基底带增生症(BZH)。BZH与纤维狭窄表型和临床有关 结果如食道僵硬和功能障碍、食物嵌塞和狭窄[1]。潜在的 推动EoE发病的免疫/炎症反应已经被广泛研究；然而，研究 在EoE中确定食道上皮细胞增殖和BZH的分子基础是稀疏的。在此，我们 分泌型FrizzledRelated Protein 1(SFRP1)在推动食道上皮细胞增殖中的关键作用 在EoE中。我们知识中的一个主要差距是sFRP1调节白细胞介素13(IL-13)- 诱导EoE大鼠食道上皮细胞增殖及补肾活血方。 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，而IL-13在食道中上调 EoE患者的活检组织，并足以改变食道上皮细胞的基因表达 活着。在初步研究中，我们发现IL-13诱导p-STAT3呈时间依赖性增加(Y705和S727) 和p-STAT6(Y641)在食道上皮细胞(EPC2-ALI)和3D原代培养的食道细胞中； 然而，IL-13诱导的食道上皮细胞增殖是STAT3依赖的。差异表达基因 (DEGS)来自EoE个体和IL-13诱导的食道活检样本的RNAseq分析 EPC2-ALI细胞富含含有STAT3结合基序的基因，并富含基因 参与上皮细胞的增殖，是促进生存的途径。我们鉴定了分泌的FrizzledRepatedProtein 1(SFRP1)，Wnt信号和细胞增殖的可溶性调节器，作为一个共同的STAT3推定靶点 和食道上皮细胞中的DEG。我们将检验IL-13-STAT3依赖的中心假设 食道上皮细胞增殖的调控是由SFRP1介导的。具体目标(SA)将定义 STAT3在IL-13诱导的食道上皮细胞表达SFRP1中的需求和SA2)定义 SFRP1在IL-13诱导的食道上皮细胞增殖中的作用 我的长期职业目标是成为一名学术背景下的独立调查员。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/转化性研究计划。拟议的研究将由一名 由专家科学家和临床医生组成的多学科团队，他们有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 从事医学研究的职业。

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.