Mechanisms of essential calcium signaling during gastric epithelial wound healing

胃上皮伤口愈合过程中必需的钙信号传导机制

• 批准号: 8886564
• 负责人: MARSHALL H MONTROSE
• 金额: $ 35.57万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8886564
• 关键词: Actins; Agonist; Anti-Inflammatory Agents; Anti-inflammatory; Biochemical; Biological Assay; CDC42 gene; Calcium Signaling; Cell Line; Cells; Cellular Structures; Cytoskeleton; Dependence; Dinoprostone; Employee Strikes; Epithelial; Epithelium; Event; Exposure to; Gases; Gastritis; Healed; Healthcare Systems; Helicobacter Infections; Helicobacter pylori; Image; In Vitro; Infection; Intervention; Lead; Learning; Lesion; Link; MYLK gene; Measures; Mediating; Microscopic; Modeling; Molecular; Monomeric GTP-Binding Proteins; Mus; NHE2; Optics; Organoids; Outcome; Pathogenesis; Pathology; Pathway interactions; Peptic Ulcer; Pharmaceutical Preparations; Phospholipase C; Process; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Protein Kinase C; Pylorus; Regulation; Role; Signal Pathway; Signal Transduction; Speed; Stomach; Surface; Technology; Testing; Therapeutic; Tight Junctions; Time; Tissues; Toxic effect; Ulcer; Work; Wound Healing; base; cell motility; citrate carrier; extracellular; healing; improved; in vivo; inhibitor/antagonist; innovation; irritation; loris; migration; pathogen; public health relevance; repaired; research study; response; rho; tool; upper GI series

 DESCRIPTION (provided by applicant): Peptic ulcer disease and upper GI irritation are a significant burden in the US healthcare system and worldwide, caused in large part by the upper GI toxicity of ingested non-steroidal anti-inflammatory drugs (NSAIDs) and the pathogen H. pylori. The rationale for our work is that improved or alternative therapeutic strategies can be based on understanding the mechanisms of gastric defense rallied against such challenges. Preliminary studies show an essential role for both intracellular and extracellular Ca2+ mobilization in gastric repair of the surface epithelium. We hypothesize that this Ca2+ mobilization is a central regulator of repair that underlies the effects of diverse agonists shown to modulate the repair process, and that this pathway can be disrupted by NSAIDs and H. pylori. The objective of this application is to use the newly identified Ca2+ signals as heralds to identif the underlying upstream and downstream mechanisms mediating gastric repair, and as a tool to investigate if NSAIDs and/or H. pylori compromise repair via these pathways. We have pioneered optical technologies that allow real-time creation of focal damage and continual quantification of repair. Our work focusing on the repair of focal lesions in vivo will be extrapolated to experiments evaluating ulceration to enhance relevance to gastric pathologies observed clinically, and to study of gastric organoids as a potentially powerful new model of gastric epithelial function. Our first aim is to identify the initiating extracellular signals in damaged tissue that stimulate Ca2+ mobilization and gastric repair. We will focus on TFF2 and prostaglandins, shown to be essential to gastric repair, but whose role in Ca2+ signaling is untested. The second aim establishes the hierarchy of Ca2+-dependent intracellular signaling pathways that are essential to allow repair. Experiments are based on preliminary findings that show phospholipase C, and protein kinase C are required for efficient gastric repair in the intact stomach. Our third aim evaluates the downstream targets of Ca2+- dependent signaling that perform the work of gastric repair. We will evaluate the role of the NHE2 Na/H exchanger, shown to be essential for gastric repair as a downstream effector of TFF2. We will also evaluate Ca2+ -dependent regulation of the actin cytoskeleton, tight junctions, and the small GTPases that regulate these cellular structures during repair. All aims focus on the role of the cyclooxygenase (COX)/prostaglandin pathway inhibited by NSAIDs, and the first and third aims additionally examine changes in the studied pathways caused by infection with H. pylori. The outcomes will provide a unique window into understanding early factors in gastric pathogenesis, which have potential to lead to new targets and new strategies for interventions that can minimize gastric damage and speed healing.

 描述（由适用提供）：消化性溃疡疾病和上胃肠道刺激是美国医疗保健系统和全球范围内的显着燃烧，这在很大程度上是由于摄入的非甾体类抗炎药（NSAID）和幽门原体H.幽门原体的上胃肠道毒性所引起的。我们工作的理由是，改进或替代治疗策略可以基于理解胃防御机制反对此类挑战的机制。初步研究表明，细胞内和细胞外Ca2+动员在表面上皮的胃修复中的重要作用。我们假设这种CA2+动员是修复的中心调节剂，它是证明可以调节修复过程的发散激动剂的作用的基础，并且该途径可以被NSAIDS和H. Pylori破坏。该应用的目的是使用新鉴定的Ca2+信号作为预言，以识别介导胃修复的上游和下游机制，并作为研究NSAID和/或H. Pylori是否通过这些途径进行修复的工具。我们拥有开创性的光学技术，可以实时创建局灶性损害和维修的连续量化。我们的工作重点是修复体内局灶性病变，将推断出评估溃疡的实验，以增强与临床观察到的胃病理学相关的相关性，并研究胃官类型作为胃内上皮功能的潜在强大新模型。我们的第一个目的是确定损害组织中刺激CA2+动员和胃修复的细胞外信号。我们将专注于TFF2和前列腺素，这对胃修复至关重要，但在CA2+信号中的作用未经测试。第二个目标建立了Ca2+依赖性细胞内信号通路的层次结构，这对于允许修复至关重要。实验基于显示磷脂酶C的初步发现，并且需要在完整摊位中有效的胃修复蛋白激酶C。我们的第三个目标评估了执行胃修复工作的Ca2+ - 依赖性信号的下游目标。我们将评估NHE2 NA/H交换器的作用，该作用对于胃修复是TFF2的下游效应器所必需的。我们还将评估肌动蛋白细胞骨架，紧密连接的Ca2+依赖性调节，以及在修复过程中调节这些细胞结构的小GTP酶。所有目标都集中在NSAID抑制的环氧合酶（COX）/前列腺素途径的作用上，第一个和第三个目标还检查了由H.幽门螺杆菌感染引起的研究IOD途径的变化。结果将为理解胃发病机理的早期因素提供一个独特的窗口，这些因素有可能导致新的目标和新策略，以最大程度地减少胃损伤和速度愈合。