Mechanisms of tissue damage leading to chronic allergic eosinophilic esophagitis

描述（由申请人提供）：嗜酸性食管炎（EE）是一种慢性过敏性疾病，患病率不断增加，诊断为酸非依赖性食管嗜酸性粒细胞增多症，每个高倍视野>15。食管上皮增生、平滑肌动力障碍和食管狭窄是EE相关特征。组织重塑与上皮变化和上皮下纤维化发生在EE和涉及TGF 21，促纤维化分子。然而，上皮和平滑肌变化的机制在很大程度上是不清楚的，更严重的EE表型的遗传因素是未知的。没有替代指标的EE。儿童必须接受反复侵入性内镜检查和组织活检的诊断和管理。我们的EE中心为研究EE的机制和了解其对临床疾病的影响提供了必要的患者人群和协作互动。我们的第一个目标是基于我们的初步研究，证明Cripto-1，一种新的蛋白质在EE中，在儿童EE患者的食管上皮中增加。我们将使用原代食管上皮细胞来了解Cripto-1如何增加食管上皮细胞增殖，以及Cripto-1是否与TGF 2家族成员一起工作。我们将确定Cripto-1在EE患者食管中的表达，与正常患者相比，并确定Cripto-1是否作为EE的生物标志物发挥作用。我们的第二个目的是基于我们目前的研究，这些研究显示了EE患者的平滑肌被产生TGF 21的肥大细胞浸润，TGF 21增加食管平滑肌细胞收缩和受磷蛋白，我们将使用原代食管平滑肌细胞，EE患者肌成纤维细胞，基因沉默技术，以及体外收缩试验的新应用来描绘TGF 21介导的收缩是否依赖于受磷蛋白。我们的第三个目的是基于我们的观察，即TGF 21启动子（C-509 T）中的功能性单核苷酸多态性（SNP）与口服局部皮质类固醇后EE的消退相关。将完成EE患者和对照的SNP分析，以确定CC基因型是否与治疗应答表型相关，TT基因型是否与更严重的EE表型相关。使用CC和TT基因型EE患者的原代细胞和染色质免疫沉淀YY-1的实验将确定在体内占领的TGF-21启动子和启动子活性EE。这些研究旨在提供创新的技术和新的治疗靶点的EE患者的理解和护理。

DESCRIPTION (provided by applicant): Eosinophilic esophagitis (EE) is a chronic allergic disease of increasing prevalence diagnosed as an acid- independent esophageal eosinophilia of >15 per high power field. Esophageal epithelial proliferation, smooth muscle dysmotility, and esophageal strictures are EE associated features. Tissue remodeling with epithelial changes and subepithelial fibrosis occur in EE and involve TGF21, a pro-fibrotic molecule. However, the mechanisms for epithelial and smooth muscle changes are largely unclear; and the genetic factors for a more severe EE phenotype are unknown. There are no surrogate markers for EE. Children must undergo repeated invasive endoscopy with tissue biopsy for diagnosis and management. Our EE center provides the essential patient population and collaborative interactions for studying the mechanisms of EE and for understanding their impact on clinical disease. Our first aim is based on our preliminary studies demonstrating that Cripto-1, a new protein in EE, is increased in the esophageal epithelium of pediatric EE patients. We will use primary esophageal epithelial cells to understand how Cripto-1 increases esophageal epithelial cell proliferation and if Cripto-1 works with TGF2 family members. We will determine the expression of Cripto-1 in the esophagus of EE patients as compared with normal patients and define if Cripto-1 functions as a biomarker for EE. Our second aim is based on our current studies showing the novel findings that the smooth muscle in EE patients is infiltrated by mast cells that produce TGF21 and that TGF21 increases both esophageal smooth muscle cell contraction and phospholamban, a new protein in EE. We will use primary esophageal smooth muscle cells, EE patient myofibroblasts, gene silencing techniques, and a novel application of an in vitro contraction assay to delineate if TGF21 mediated contraction relies on phospholamban. Our third aim is based on our observation that a functional single nucleotide polymorphism (SNP) in the TGF21 promoter (C-509T) is associated with resolution of EE following swallowed topical corticosteroids. SNP analysis on EE patients and controls will be completed to define if the CC genotype associates with a therapy responsive phenotype and if the TT genotype associates with a more severe EE phenotype. Experiments using primary cells from EE patients of CC and TT genotype and chromatin immunoprecipitation for YY-1 will determine in vivo occupation of the TGF21 promoter and promoter activity in EE. These studies are designed to provide innovative techniques and new therapeutic targets for the understanding and care of EE patients.