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)是一种由抗原引起的慢性过敏性疾病，可引起临床症状 呕吐，吞咽困难，儿童发育不良。由于慢性嗜酸性炎症，未经治疗的 或者治疗无效的EoE食道变得僵硬、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 在改变食道成纤维细胞和平滑肌功能中的不可或缺的作用。促纤维化因子可以成为 陷于ECM和基质刚性可以改变结构细胞的功能。然而，我们目前缺乏一种 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 成纤维细胞可以反映EoE的疾病严重程度或预测食道狭窄的风险。此外，还有 没有简单的方法来衡量对抗重塑化合物的治疗反应-这是患者护理和 药物开发。这些问题为最佳病人护理创造了巨大的知识差距。同时发炎 启动组织重塑，它既不是食道功能障碍的唯一传播因子，也不是最好的预测因素 病理性纤维化的标志物。在本应用中，我们建议使用原代人食道成纤维细胞 不同程度EoE患者与来自正常食道的ECM之间的相互作用 以及成纤维细胞，以破译驱动成纤维细胞功能变化的最相关分子。至 要做到这一点，我们将使用有针对性的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评估潜在的抗重塑化合物阻断细胞外基质的能力 诱导成纤维细胞功能改变，从而打造潜在的个性化抗逆转录病毒药物平台 纤维状化合物。我们将把我们的体外研究结果与体内蛋白质表达研究以及长期的 使用我们的表型良好的纵向EoE队列的术语疾病轨迹。我们假设这些研究 将确定改变EoE进程并作为新的治疗靶点的新的相关蛋白质 用于治疗疾病并发症。

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.