Mechanisms of tissue damage leading to chronic allergic eosinophilic esophagitis

描述（由申请人提供）：嗜酸性粒细胞性食管炎（EE）是一种慢性过敏性疾病，发病率越来越高，诊断为每高倍视场bbbb15的非酸依赖性食管嗜酸性粒细胞增多。食管上皮增生、平滑肌运动障碍和食管狭窄是EE的相关特征。伴有上皮改变和上皮下纤维化的组织重塑发生在EE中，并涉及促纤维化分子TGF21。然而，上皮和平滑肌变化的机制在很大程度上尚不清楚；更严重的EE表型的遗传因素尚不清楚。情感表达没有替代标记物。儿童必须反复进行侵入性内窥镜检查和组织活检以进行诊断和治疗。我们的情感表达中心为研究情感表达的机制和理解它们对临床疾病的影响提供了必要的患者群体和协作互动。我们的第一个目标是基于我们的初步研究表明，EE中的一种新蛋白Cripto-1在儿科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的增殖，以及Cripto-1是否与TGF2家族成员一起起作用。我们将测定EE患者与正常患者食管中Cripto-1的表达，并确定Cripto-1是否作为EE的生物标志物。我们的第二个目标是基于我们目前的研究显示，EE患者的平滑肌被产生TGF21的肥大细胞浸润，TGF21增加食管平滑肌细胞收缩和磷酸化蛋白（EE中的一种新蛋白）。我们将使用原代食管平滑肌细胞、EE患者肌成纤维细胞、基因沉默技术和一种新型的体外收缩实验来描述TGF21介导的收缩是否依赖于磷蛋白。我们的第三个目标是基于我们的观察，TGF21启动子（C-509T）中的功能性单核苷酸多态性（SNP）与吞咽局部皮质类固醇后EE的溶解有关。将完成对EE患者和对照组的SNP分析，以确定CC基因型是否与治疗反应表型相关，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y -1的实验将确定TGF21启动子在EE中的体内占据和启动子活性。这些研究旨在为理解和护理情感表达患者提供创新技术和新的治疗靶点。

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.