Probing the Mechanisms of Fibroblast-Extracellular Matrix Interactions to Assess Disease Severity in Allergic Eosinophilic Esophagitis

项目摘要 嗜酸性食管炎（EoE）是一种慢性、抗原驱动的过敏性疾病， 呕吐、吞咽困难和儿童发育不良。由于慢性嗜酸性粒细胞性炎症， 或治疗无反应的EoE食管变得僵硬、狭窄和运动障碍，导致食物嵌塞。 组织重塑包括组织学上皮下纤维化、血管生成和平滑肌肥大， 是EoE并发症的潜在机制。上皮下细胞外基质（ECM）在肿瘤的发生发展中起着重要的作用。 在改变食管成纤维细胞和平滑肌功能中起重要作用。促纤维化因素可以成为 被困在ECM和基质刚性可以改变结构细胞功能。然而，我们目前缺乏一个 了解EoE ECM中纤维化的主要分子驱动因素，以及ECM之间的相互作用 成纤维细胞可反映EoE患者疾病的严重程度或预测食管狭窄的风险。此外，还有 没有简单的方法来衡量抗重塑化合物的治疗反应，这是患者护理的一个障碍， 药物开发这些问题造成了最佳患者护理的重大知识差距。虽然炎症 启动组织重塑，它既不是食管功能障碍的唯一传播者，也不是最好的预测因子。 病理性纤维化的标志物。在本申请中，我们提出使用来自人食管成纤维细胞的原代人食管成纤维细胞。 不同严重程度EoE和正常食管的患者，以了解ECM之间的相互作用， 和成纤维细胞，以便破译驱动成纤维细胞功能变化的最相关分子。到 为了实现这一点，我们将使用靶向蛋白质分析和无偏见的蛋白质组学方法。然后我们计划 使用我们的实验方法来评估潜在的抗重塑化合物阻断ECM的能力， 诱导成纤维细胞功能的变化，从而创造了一个潜在的个性化药物平台， 纤维化化合物我们将把我们的体外研究结果与体内蛋白质表达研究和长期研究结果结合起来， 长期疾病轨迹使用我们的良好表型纵向EoE队列。我们假设这些研究 将确定新的和相关的蛋白质，改变EoE的过程，并作为新的治疗靶点 治疗疾病并发症

Project Summary Eosinophilic esophagitis (EoE) is a chronic, antigen driven allergic disease that causes clinical symptoms of vomiting, trouble swallowing, and poor growth in children. Due to chronic eosinophilic inflammation, the untreated or therapy unresponsive EoE esophagus becomes rigid, narrowed, and dysmotile, resulting in food impactions. Tissue remodeling includes histologic subepithelial fibrosis, angiogenesis and smooth muscle hypertrophy and is the underlying mechanism for EoE complications. The subepithelial extracellular matrix (ECM) plays an integral role in changing esophageal fibroblast and smooth muscle function. Pro-fibrotic factors can become trapped in the ECM and matrix rigidity can alter structural cell functions. However, we currently lack an understanding of the main molecular drivers of fibrosis in the EoE ECM and if the interactions between the ECM and fibroblasts can reflect disease severity or predict the risk of esophageal narrowing in EoE. Further, there are no easy ways to gauge therapeutic response to anti-remodeling compounds—a hinderance for patient care and drug development. These issues create significant knowledge gaps for optimal patient care. While inflammation initiates tissue remodeling, it is neither the sole propagator of esophageal dysfunction nor the best predictive marker of pathological fibrosis. In this application we propose to use primary human esophageal fibroblasts from patients with varying severities of EoE and from normal esophagi to understand the interaction between the ECM and fibroblasts in order to decipher the most relevant molecules driving changes in fibroblast function. To accomplish this, we will use both targeted protein analysis and an unbiased proteomic approach. We then plan to use our experimental approach to assess the ability of potential anti-remodeling compounds to block ECM induced changes in fibroblast function, thereby creating a potential personalized medicine platform for anti- fibrotic compounds. We will align our in vitro findings with in vivo protein expression studies and with the long- term disease trajectory using our well phenotyped longitudinal EoE cohort. We hypothesize that these studies will determine novel and relevant proteins that alter the course of EoE and function as new therapeutic targets for disease complications.