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)是一种慢性变态反应性疾病，发病率不断上升，诊断为每高倍视野15-gt；15的酸非依赖性食道嗜酸性粒细胞增多症。食道上皮细胞增殖、平滑肌运动障碍和食道狭窄是EE的相关特征。EE的组织重塑伴随上皮样改变和上皮下纤维化，并涉及促纤维化分子TGF21。然而，上皮和平滑肌改变的机制很大程度上还不清楚；更严重的EE表型的遗传因素也是未知的。EE没有代理标记。儿童必须接受反复的侵入性内窥镜检查和组织活检进行诊断和治疗。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基本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在儿童EE患者的食道上皮细胞中增加。我们将使用原代食道上皮细胞来了解Cripto-1如何促进食道上皮细胞的增殖，以及Cripto-1是否与TGF2家族成员一起工作。我们将检测Cripto-1在EE患者和正常患者的食道中的表达，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，这些新发现表明EE患者的平滑肌被产生TGF21的肥大细胞渗透，并且TGF21既增加了食道平滑肌细胞的收缩，也增加了EE中的一种新蛋白-磷蛋白。我们将使用原代食道平滑肌细胞、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，即TGF21启动子(C-509T)上的功能性单核苷酸多态(SNP)与吞咽局部皮质类固醇后EE的解决相关。将完成对EE患者和对照的SNP分析，以确定CC基因是否与治疗反应表型相关，TT基因是否与更严重的EE表型相关。使用CC和TT型EE患者的原代细胞和染色质免疫沉淀检测YY-1的实验将确定TGF21启动子在EE中的体内占有率和启动子活性。这些研究旨在为EE患者的理解和护理提供创新的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.