Maturational Characterization of Human Esophageal Fibroblasts in EoE

描述（由申请方提供）：嗜酸性粒细胞性食管炎（EoE）是一种过敏性疾病，其特征为嗜酸性粒细胞的食管浸润，从儿童期到成年期诊断。食管纤维化是EoE最严重的并发症，导致吞咽困难和食管食物嵌塞，开始于生命的第二个十年。阐明导致纤维化的机制对于理解EoE的自然史至关重要。我们的初步数据表明，原代EoE食管上皮细胞（EPCs）和原代胎儿食管成纤维细胞（FEF 3）之间的串扰增强FEF 3的促纤维化细胞因子的表达。然而，我们的模型受到胎儿成纤维细胞使用的限制，胎儿成纤维细胞在功能上不同于儿科和成人成纤维细胞。迄今为止，食管癌的特征 从未描述过跨越儿科和成人EoE群体的成纤维细胞。我们的总体假设是，EoE受试者的食管成纤维细胞与非EoE受试者的成纤维细胞在表型上不同，并且EoE成纤维细胞的致突变特征在人类成熟过程中增强。使用原代食管成纤维细胞，我们将表征成纤维细胞的细胞因子反应和收缩性，比较年龄匹配的EoE和非EoE受试者之间的反应（目的1a）。将检查年龄为1-9岁、10-17岁和成人的EoE受试者中纤维化反应的EoE相关差异（目的1b）。我们将确定成纤维细胞对生理相关上皮刺激（包括酸/胆汁和EoE触发物（食物抗原））背景下上皮衍生因子的反应（目的2a）。最后，这些刺激对纤维化的影响将使用新的器官型细胞培养模型的“活”食管组织，构建从原代EPCs和食管成纤维细胞系（目标2b）。

DESCRIPTION (provided by applicant): Eosinophilic esophagitis (EoE) is an allergic disease characterized by esophageal infiltration of eosinophils, diagnosed from childhood through adulthood. Esophageal fibrosis is the most serious complication of EoE, leading to dysphagia and esophageal food impaction starting in the second decade of life. Elucidating the mechanisms which lead to fibrosis is critical to understanding the natural history of EoE. Our preliminary data suggests that cross-talk between primary EoE esophageal epithelial cells (EPCs) and primary fetal esophageal fibroblast cells (FEF3) enhances FEF3 expression of profibrotic cytokines. However, our model is limited by its use of fetal fibroblasts, which are functionally distinct from pediatric and adult fibroblasts. To date, characterization of esophageal fibroblasts spanning pediatric and adult EoE populations has never been described. Our overall hypothesis is that esophageal fibroblasts from EoE subjects are phenotypically distinct from fibroblasts from non-EoE subjects, and that fibro genic characteristics of EoE fibroblasts are enhanced during human maturation. Using primary esophageal fibroblasts, we will characterize fibroblast cytokine responses and contractility, comparing responses between age-matched EoE and non-EoE subjects (Aim 1a). Age-related differences in fibrotic responses in EoE subjects ages 1-9 years, 10-17 years, and adults will be examined (Aim 1b). We will determine fibroblast responses to epithelial-derived factors in the context of physiologically relevant epithelial stimuli including acid/bile and EoE triggers (food antigens) (Aim 2a). Finally, the effects of these stimuli upon fibrosis will be examined using novel organotypic cell culture models of "living" esophageal tissue, constructed from primary EPCs and esophageal fibroblast cell lines (Aim 2b).