Novel minimally invasive assessment of gastrointestinal inflammation in food alle

描述(申请人提供)：嗜酸性食管炎(EE)是一种日益公认的胃肠道疾病，约占儿童和成人吞咽困难和食物嵌塞病例的50%。慢性嗜酸性粒细胞炎症在EE中的长期后果包括食道重塑、上皮下纤维化、以及食道狭窄和狭窄，因此强调了这种疾病的重要性。翻译研究表明，90%的EE患者对饮食干预有反应，支持体液(IgE)和/或细胞介导的食物过敏的作用。由于RAST检测对识别致病食物变应原(S)的特异性较差，而SPTS仍是识别相关食物变应原的最佳检测方法，因此Ig E介导的肥大细胞(或嗜碱性细胞)反应在启动食管炎性级联反应中的作用仍不确定。EE的标准护理包括最初的内窥镜检查和活检以获取食道组织以确定嗜酸性粒细胞的数量，=15/HPF为诊断。到目前为止，还没有血清学或粪便分析提供与疾病进展与缓解的组织学证据相关的持久发现。重复内窥镜检查和活组织检查的作用仍在争论中，因为成本效益比尚不清楚，而且存在风险。我们假设并提供了初步结果，即Enterotest(Tm)，一种最初用于检测肠道贾第鞭毛虫的基于字符串的测试，可以用于在蛋白质和mRNA水平上评估食管炎(称为食管串测试或EST)。该项目的目标是：(1)测试EST作为一种新的、微创、廉价、灵敏和特异的方法来测量EE患者的食管炎，特别是组织嗜酸粒细胞、细胞因子和趋化因子的变化，以及CD23、FceRI和类胰蛋白酶的水平，以及(2)使用EST来监测营养或皮质类固醇治疗后的疾病状态。我们提出了两个完整的假设：(1)EST检测到的管腔炎症介质与食道活检确定的EE患者的粘膜炎症相关，(2)EST可用于疾病治疗的食管炎症的微创临床评估。目的1应用EST技术研究EE患者食管腔内微环境的炎症反应，通过检测嗜酸性粒细胞表达的炎性介质(嗜酸性粒细胞颗粒蛋白)，确定其与EE病组织病理学特征的关系。目的比较血浆、食道粘膜(活检)和管腔(EST)CD23、FceRI、类胰蛋白酶、Th1和Th2细胞因子、嗜酸细胞趋化蛋白-3和嗜酸性粒细胞颗粒蛋白生物标志物的定量测定，以确定治疗对EE患者食管炎性反应的影响。这些研究将开始验证EST作为监测EE中食管炎的微创诊断工具，CD23/FceRI/IgE的参与，以及使用EST代替重复的内窥镜和活检来确定治疗的组织病理学反应的有效性。(与公共卫生有关)食物过敏性疾病(包括嗜酸性胃肠道疾病，如嗜酸性食管炎)目前约占美国人口的4%至6%。对食道过敏症患者的护理是有限的，因为需要初始和重复的内窥镜和活组织检查来诊断和评估疾病缓解或进展方面的治疗效果。这项应用侧重于通过使用现有的微创技术Enterotest(Tm)来验证一种新的方法-食管串试验，以分析食管炎，并使用这一新的测试来评估治疗对食管炎的影响以及IgE介导的途径在嗜酸性食管炎发病机制中的参与。这些研究的结果将通过限制侵入性内窥镜检查的数量和护理所需的活检程序，并通过增加对嗜酸性食管炎和其他食物过敏性胃肠道疾病的发病机制的全面了解，来改善食物过敏和嗜酸性食管炎患者的生活质量。

DESCRIPTION (provided by applicant): Eosinophilic esophagitis (EE) is an increasingly recognized gastrointestinal disease that accounts for ~50% of pediatric and adult cases of dysphagia and food impaction. Long-term consequences of chronic eosinophilic inflammation in EE include esophageal remodeling, with subepithelial fibrosis, and esophageal narrowing and strictures, thus emphasizing the importance of this disease. Translational studies show that >90% of EE patients respond to dietary interventions, supporting a role for humoral (IgE) and/or cell-mediated food allergy. The role of IgE-mediated mast cell (or basophil) responses in initiating the esophageal inflammatory cascade remains uncertain since RAST testing alone shows poor specificity for identifying the offending food allergen(s), whereas SPTs remain the best available test to identify relevant food allergens. Standard of care for EE includes initial endoscopy with biopsy for obtaining esophageal tissue to determine the numbers of eosinophils, =15/hpf being diagnostic. No serological or stool analyses to date provide durable findings that correlate with histological evidence of disease progression vs. remission. The role of repeated endoscopy and biopsy is under debate, since the cost-benefit ratio is unknown and entails risks. We hypothesize and present preliminary results that the Enterotest,(tm) a string-based test first used for detection of intestinal Giardia, can be used to assess esophageal inflammation at the protein and mRNA levels (termed the Esophageal String Test or EST). The project's objectives are to: (1) test the efficacy of the EST as a novel, minimally invasive, inexpensive, sensitive and specific method for measuring esophageal inflammation in EE patients, particularly tissue eosinophilia, cytokine and chemokine profiles, and levels of CD23, FceRI and tryptase, and (2) use the EST to monitor disease status following nutritional or corticosteroid treatments. We propose two integrated hypotheses that: (1) luminal inflammatory mediators detected using EST correlate with mucosal inflammation as determined by esophageal biopsy in patients with EE, and (2) the EST can be used as a minimally invasive clinical assessment of esophageal inflammation for disease management. Aim 1 will characterize the inflammatory response in the esophageal luminal microenvironment of EE patients using the EST, performing measurements of specific eosinophil-expressed inflammatory mediators (eosinophil granule proteins), to determine their associations with the histopathologic features of EE disease. Aim 2 will determine the impact of treatment on the esophageal inflammatory response in EE patients comparing quantitative measurements of plasma, esophageal mucosa (biopsy) and luminal (EST) CD23, FceRI, tryptase, Th1 and Th2 cytokines, eotaxin-3, and eosinophil granule protein biomarkers to correlate changes with treatment-induced responses. These studies will begin to validate the EST as a minimally invasive diagnostic tool for monitoring esophageal inflammation in EE, the participation of CD23/FceRI/IgE, and the efficacy of using the EST to determine histopathologic responses to treatment in lieu of repeated endoscopies and biopsies. (PUBLIC HEALTH RELEVANCE) Food allergic diseases (including eosinophilic gastrointestinal diseases such as eosinophilic esophagitis) currently afflict an estimated 4-6% of the United States population. Care of patients with esophageal allergies is limited because of the need for initial and repeated endoscopies and biopsies for diagnosis and evaluation of the effectiveness of treatment in terms of disease remission or progression. This application focuses on studies validating a novel method, the Esophageal String Test, for analyzing esophageal inflammation through the use of minimally invasive existing technology, the Enterotest(tm), and using this novel test to evaluate the effects of treatment on esophageal inflammation and the participation of IgE-mediated pathways in the pathogenesis of eosinophilic esophagitis. Results from these studies will improve the quality of life for patients with food allergies and eosinophilic esophagitis by limiting the number of invasive endoscopy with biopsy procedures required for their care, and by increasing overall understanding of the pathogenesis of eosinophilic esophagitis and other food allergic gastrointestinal diseases.