Lysyl oxidase induced esophageal remodeling in eosinophilic esophagitis

项目摘要 嗜酸性食管炎(EoE)是一种以持续性过敏为特征的慢性衰弱疾病。 随着时间的推移会导致纤维化和狭窄的炎症。虽然它的特点是强烈的嗜酸性粒细胞和 肥大细胞浸润、病理性组织重塑事件导致不可逆转的纤维化和显著的临床 症状。这些重塑事件包括上皮基底细胞增生和上皮下纤维化。我们的 以前的研究已经发现赖氨酰氧化酶(LOX)，这是一种已知的交联胶原蛋白和增强组织的酶 僵硬是病理性食道重塑的关键调节因子。LOX在食道中强势表达 活动期EoE炎症患者的粘膜，尤其是纤维化者。在体外和体内的健壮 数据表明，转化生长因子信号在食道上皮细胞中促进脂氧合酶的产生，进而促进 上皮细胞(基底细胞增生)和成纤维细胞间隔的重塑。因此，我们寻求 明确上皮转化生长因子和脂氧合酶促进食道重塑的机制 利用这些机制进行临床应用。我们假设上皮性转化生长因子-LOX轴驱动基底 EoE中的细胞增生和纤维狭窄。在目标1中，我们将评估上皮细胞的自主作用 评估上皮转化生长因子如何在3D上皮细胞培养和培养中驱动LOX诱导的基底细胞增殖 染色质免疫沉淀分析。在目标2中，我们将评估上皮细胞的非细胞自主效应 通过评估转化生长因子和脂氧合酶对成纤维细胞活化和组织硬度的影响。最后，在目标3中，我们 将通过进行临床前的目的验证来评估这一途径的治疗和预后潜力 抗转化生长因子抗体治疗EoE炎症及验证EoE中LOX表达的研究 病人组织。这项研究的数据将为未来针对组织重塑的治疗以及工作提供信息 有助于在残疾发作之前识别纤维化疾病患者。 这项创新和假设驱动的研究得到了PI生成的强大初步数据的支持 而她得到了NIH职业K08奖的支持。PI独特地准备通过以下方式实现这些目标 她之前在研究上皮成纤维细胞串扰、EoE中LOX和3D机制方面的记录 上皮细胞培养和共培养模型。

Project Summary Eosinophilic esophagitis (EoE) is a chronic, debilitating disorder characterized by persistent allergic inflammation that over time leads to fibrosis and stricture. While it is characterized by intense eosinophil and mast cell infiltration, pathologic tissue remodeling events lead to irreversible fibrosis and significant clinical symptoms. These remodeling events include epithelial basal cell hyperplasia and subepithelial fibrosis. Our previous work has implicated lysyl oxidase (LOX), an enzyme known to cross-link collagen and potentiate tissue stiffness, as a key mediator of pathologic esophageal remodeling. LOX is robustly expressed in the esophageal mucosa of patients with active EoE inflammation, particularly in those with fibrosis. Robust in vitro and in vivo data suggest that TGF signaling in esophageal epithelium enhances LOX production, which in turn, promotes remodeling in both the epithelial (basal cell hyperplasia) and fibroblast compartments. We therefore seek to define the mechanisms whereby epithelial TGF and LOX promote remodeling in the esophagus and further exploit these mechanisms for clinical application. We hypothesize that the epithelial TGF-LOX axis drives basal cell hyperplasia and fibrostenosis in EoE. In Aim 1, we will evaluate the cell autonomous effects of epithelial LOX, evaluating how epithelial TGF drives LOX-induced basal cell hyperplasia in 3D epithelial culture and chromatin immunoprecipitation assays. In Aim 2, we will evaluate the non-cell autonomous effects of epithelial TGF and LOX by evaluating their effects on fibroblast activation and tissue stiffness. And finally, in Aim 3 we will evaluate the therapeutic and prognostic potential of this pathway by performing pre-clinical proof of purpose studies, treating mice with EoE inflammation with anti-TGF antibody and validating LOX expression in EoE patient tissue. Data from this study will inform future therapies targeting tissue remodeling as well as work towards identifying patients with fibrotic disease prior to the onset of disability. This innovative and hypothesis-driven study is backed by strong preliminary data generated by the PI while she was supported by an NIH career K08 award. The PI is uniquely poised to accomplish these aims with her previous track record in investigating mechanisms of epithelial-fibroblast crosstalk, LOX in EoE, and 3D epithelial culture and co-culture models.