Molecular Mechanisms of Epithelial Contribution to Esophageal Inflammation and Tissue Repair

项目总结 这项建议的首要目标是了解上皮对食道的作用。 核内关键炎性介质抑制物的作用机制研究 KappaB-KK亚基β(IKKβ)。这项建议的意义在于1)流行的 食道紊乱，这在美国和全世界都是一个沉重的负担，2)中央 上皮性IKKβ信号在多种生物学过程中的调节作用，如炎症， 多种组织和细胞类型中的免疫、细胞存活和细胞生长。私家侦探是一名早期调查员 世卫组织是上皮炎症和损伤、炎症介质、疾病动物模型以及 上皮生物学，并由一支精湛的研究团队提供支持，并由专家合作者补充。这里, 我们将利用新的小鼠模型和补充的体外系统，利用3D培养系统 为了验证这样一种假设，即激活上皮细胞IKKβ信号会产生一种促炎、促炎症- 增殖的，促进血管生成的环境，使平衡趋向于发展食道疾病，如 鳞状细胞不典型增生和癌症。为了探索这些过程，我们将进行三个相互关联的 明确的目标。在目标1中，我们将确定STAT3激活和IKKβ信号的功能相互作用。 食道上皮细胞的增殖反应。这将使用两个活动的小鼠进行 IKKB和STAT3缺失，以及三维(3D)小鼠食道器官。在目标2中，我们将定义 上皮细胞IKKβ信号转导通路在上皮-内皮细胞相互作用中的作用 状态为食道发育不良。在这里，我们将利用一种新的具有食道上皮细胞的转基因小鼠模型 用4-硝基喹啉-1-氧化物(4-NQO)处理IKKβ缺失，并进行三维血管网络形成实验。在AIM 3，我们将探讨上皮IKKβ信号在食道上皮炎症反应中的作用。 细胞从正常状态过渡到食道异型增生。这将进行新的转基因 4-NQO治疗的小鼠食道上皮IKKβ缺失模型。这些相辅相成 方法将确认和扩展我们的初步数据和我们最近发表在 胃肠病学。此外，拟议的研究将得到优秀的合作者的支持。 环境和特殊的资源和设施，可供私人投资。我们预计这些研究 将提供对调节正常食道上皮内稳态的因素的洞察， 微环境，以及在食道疾病中被破坏的途径，包括良性和恶性。

PROJECT SUMMARY The overarching goal of this proposal is to understand the contribution of the epithelium to esophageal inflammation and tissue injury through mechanistic studies of the key inflammatory mediator inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ). The significance of this proposal lies in 1) the prevalence of esophageal disorders, which are a significant burden in the U.S. and throughout the world and 2) the central role of epithelial IKKβ signaling in the regulation of diverse biological processes, such as inflammation, immunity, cell survival, and cell growth in numerous tissues and cell types. The PI is an early-stage investigator who is an expert in epithelial inflammation and injury, inflammatory mediators, animal models of disease, and epithelial biology, and is supported by a superb research team, complemented by expert collaborators. Here, we will take advantage of new mouse models and complementary in vitro systems utilizing 3D culture system to test the hypothesis that activation of epithelial IKKβ signaling produces a pro-inflammatory, pro- proliferative, pro-angiogenic milieu that tips the balance towards the development esophageal diseases such as squamous cell dysplasia and cancer. To explore these processes, we will undertake three interrelated Specific Aims. In Aim 1, we will determine the functional interplay of STAT3 activation and IKKβ signaling in the proliferative response of esophageal epithelial cells. This will be undertaken using mice with both active IKKb and STAT3 deletion, and three-dimensional (3D) mouse esophageal organoids. In Aim 2, we will define the role of epithelial IKKβ signaling in epithelial-endothelial cell interactions during the transition from a normal state to esophageal dysplasia. Here, we will utilize a novel transgenic mouse model with esophageal epithelial IKKβ deletion treated with 4-Nitroquinoline 1-oxide (4-NQO), and 3D vascular network formation assay. In Aim 3, we will investigate the role of epithelial IKKβ signaling in the inflammatory response of esophageal epithelial cells during the transition from a normal state to esophageal dysplasia. This will be undertaken new transgenic mouse model of esophageal epithelial IKKβ deletion that is treated with 4-NQO. These complementary approaches will confirm and extend the findings from our Preliminary Data and from our recent publication in Gastroenterology. Moreover, the proposed research will be supported by the superb and collegial intellectual environment and the exceptional resources and facilities available to the PI. We anticipate that these studies will provide insight into the factors that regulate normal esophageal epithelial homeostasis, the microenvironment, and the pathways that are disrupted in esophageal diseases, both benign and malignant.