Regulation of epithelial barrier

上皮屏障的调节

• 批准号: 10289802
• 负责人: STELLA ALIMPERTI
• 金额: $ 45.63万
• 依托单位: ADA SCIENCE AND RESEARCH INSTITUTE LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-10 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10289802
• 关键词: Ablation; Address; Affect; Agonist; Animal Experiments; Bacteria; Behavior; Binding; Biological Models; Cell Nucleus; Cell membrane; Cells; Connective Tissue; Cues; Cytoplasm; Data; Devices; Disease; E-Cadherin; Epithelial; Epithelial Attachment; Escherichia coli; Event; Experimental Models; FOXO1A gene; Fusobacterium nucleatum; Genes; Gingiva; Homeostasis; Human; Implant; In Vitro; Inflammation; Inflammatory; Link; Maintenance; Measures; Mechanical Stimulation; Mechanical Stress; Mediating; Metals; Mouth Diseases; Mus; Oral; Patients; Periodontitis; Process; RNA Interference; Regulation; Regulatory Pathway; Research; Role; Signal Transduction; Site; Surface; System; TLR4 gene; Testing; Tight Junctions; Tissues; Titanium; base; beta catenin; improved; in vivo; insight; interstitial; keratinocyte; knock-down; mechanical force; mechanical pressure; nonhuman tissue; novel; oral bacteria; oral cavity epithelium; peri-implantitis; pressure; spatiotemporal

Project summary Periodontitis and peri-implantitis are oral diseases characterized by loss of oral homeostasis and induction of inflammation. Although downstream inflammatory events that occur in connective tissue have been well recognized, the regulatory pathways involved in epithelial barrier function in these diseases has not been explored in detail. In this study, we aim to identify key spatiotemporal mechanisms that control epithelial barrier function. Although it is recognized that epithelial barrier function is critical it's regulation is not well understood. To address this, in vivo and in vitro experimental models, such as Epi-oral device, have been developed. Based on Prel Data we will investigate the role of FOXO1, E-cadherin and β−catenin in the maintenance and loss of barrier function through bacteria induced signaling. Preliminary Data demonstrate that pressure enhances epithelial barrier and directs E-cadherin/β−catenin to cell membranes and FOXO1 to the cytoplasm. E.coli LPS, a TLR4 agonist, reverses this behavior and directs FOXO1 and β−catenin to the nucleus and from cell membranes E-cadherin. These data serve as the basis to mechanistically investigate the role of mechanical pressure on promoting epithelial integrity and its disruption by E.coli LPS and F. nucleatum, an important oral bacterium, by use of the Epi-oral platform (Aim1). Next, we will investigate how epithelial attachment to titanium may affect barrier function in studies that will give insight into processes that are important in peri-implantitis. Specifically, we will examine how the interaction of keratinocytes-titanium, as underlying matrix, regulates keratinocytes in the presence of pressure or E.coli LPS and F. nucleatum (Aim2). Finally, we will investigate how FOXO1, β−catenin and E-cadherin are modulated in healthy tissue compared to inflamed tissue in vivo. Animal experiments will involve mechanistic studies to examine the role of upstream events in barrier function by lineage specific deletion of FOXO1 and TLR4 in keratinocytes in vivo, to examine dysregulation of β−catenin, E-cadherin and the formation of tight junctions. Similar studies will be examined in human tissue from non- inflamed gingiva, and gingiva from patients with periodontitis and peri-implantitis sites to determine if similar dysregulation occurs in these disease processes (Aim3). Given the recognized fundamental importance of the complexity in oral diseases, these studies may pave the way to identify novel targets for treatments against periodontitis and peri-implantitis.

项目摘要 牙周炎和种植体周围炎是口腔疾病，其特征在于口腔内稳态的丧失和诱导牙周炎。 炎症尽管发生在结缔组织中的下游炎症事件已经被很好地 虽然已经认识到，在这些疾病中涉及上皮屏障功能的调节途径尚未被发现， 详细探索。在这项研究中，我们的目的是确定关键的时空机制，控制上皮屏障， 功能虽然认识到上皮屏障功能是至关重要的，但其调节并不很清楚。 为了解决这个问题，已经开发了体内和体外实验模型，例如Epi-oral装置。基于 在Prel数据中，我们将研究FOXO 1、E-cadherin和β-catenin在维持和丢失细胞中的作用。 屏障功能通过细菌诱导的信号传导。初步数据表明，压力增加 上皮屏障，并将E-钙粘蛋白/β-连环蛋白引导至细胞膜，FOXO 1引导至细胞质。大肠杆菌LPS， 一种TLR 4激动剂，逆转这种行为并将FOXO 1和β-连环蛋白从细胞膜引导至细胞核 E-钙粘蛋白。这些数据作为基础，机械地研究机械压力对 促进上皮完整性和其被大肠杆菌LPS和F. nucleatum是一种重要的口腔细菌， 使用Epi-oral平台（Aim 1）。接下来，我们将研究上皮细胞如何附着在钛上， 影响屏障功能的研究，将深入了解种植体周围炎的重要过程。 具体来说，我们将研究如何相互作用的角质形成细胞钛，作为底层基质，调节 角质形成细胞在压力或大肠杆菌LPS和F.核质（Aim 2）。最后，我们将调查 与体内炎症组织相比，FOXO 1、β-连环蛋白和E-钙粘蛋白在健康组织中是如何调节的。 动物实验将涉及机制研究，以检查上游事件在屏障功能中的作用 通过体内角质形成细胞中FOXO 1和TLR 4的谱系特异性缺失，以检查β-连环蛋白的失调， 钙粘蛋白与紧密连接的形成。类似的研究将在来自非- 发炎的牙龈，以及牙周炎和种植体周围炎患者的牙龈，以确定 在这些疾病过程中发生失调（Aim 3）。鉴于公认的基本重要性， 由于口腔疾病的复杂性，这些研究可能为确定新的治疗靶点铺平道路， 牙周炎和种植体周围炎。