Crohns disease, COX2, tissue repair, and the resolution of inflammation

克罗恩病、COX2、组织修复和炎症消退

• 批准号: 9152999
• 负责人: David Meriwether
• 金额: $ 3.71万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-18 至 2017-09-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9152999
• 关键词: Abdominal colic; Ablation; Accounting; Animal Genetics; Animal Model; Arachidonic Acids; Attenuated; Biological Assay; CD7 gene; Cell Differentiation process; Cell Proliferation; Cholates; Cholic Acids; Chronic; Collection; Conditioned Culture Media; Coxibs; Crohn' s disease; Data; Development; Diet; Digestive System Disorders; Dinoprostone; Disease; Electrical Resistance; Enzymes; Epithelial; Equilibrium; Etiology; Feedback; Flare; Gastrointestinal tract structure; Genes; Genetic Polymorphism; Genetic Predisposition to Disease; Genetic study; Goals; Health; Heart; Human; Immune; Immune response; Immune system; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Infiltrate; Inflammatory Response; Interleukin-10; Intestines; Investigation; Kinetics; Lead; Link; Lipids; LoxP-flanked allele; Measurement; Measures; Mediating; Methods; Modeling; Molecular; Morbidity - disease rate; Mus; Mutation; Myelogenous; Nature; Pathogenesis; Pathology; Pathway interactions; Pattern; Pattern recognition receptor; Peptidoglycan; Phenotype; Predisposition; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Prostaglandins I; Proteomics; Regulation; Relapse; Reporting; Resistance; Resolution; Risk; Role; STAT3 gene; Signal Pathway; Signal Transduction; Stem cells; Structure; Susceptibility Gene; Symptoms; System; Techniques; Thromboxanes; Time; Tissues; Ulcerative Colitis; Uncertainty; Work; case-by-case basis; cell type; commensal microbes; crypt cell; cyclooxygenase 2; cytokine; cytotoxic; genetic association; in vivo; insight; intestinal epithelium; macrophage; mouse model; novel; pathogen; research study; tissue repair; transcription factor

 DESCRIPTION (provided by applicant): COX2 is a genetic susceptibility locus with respect to Crohn's disease (CD), but the nature of the link between COX2 and CD is unclear. Mice deficient in COX2 develop Crohn's disease-like inflammation when challenged with a cholate-containing diet. This project will investigate the mechanism behind the pathology in this model in order to gain insight into the etiology of CD. Pilot data implicates COX2 in both intestinal epithelial barrier integrity and the resolution of inflammation in this model, such that COX2 deficiency appears to inhibit epithelial tissue repair while also prolonging the inflammatory response. It is thus hypothesized that in the absence of COX2, a positive feedback loop develops between altered barrier integrity and prolonged inflammation, with each reinforcing the other. Aim 1 seeks to characterize the direct effects of cholate and COX2 on barrier integrity in this model. Time course experiments, in vitro studies, and bacterial ablation will all be employed to isolate the direct effects of cholate on barrier integrity from the confounding effects of inflammation. Barrier integrity will be partially characterized through the use of in vivo translocation assays together with measurements of trans-epithelial electrical resistance within a Ussing chamber. Aim 2 further investigates this hypothesis by studying the effect of COX2 on intestinal stem cell (ISC) proliferation and differentiation within a primary enteroid culture system. Preliminary data indicates that COX2 inhibition within crypts limits crypt expansion, while both exogenous PGE2 and macrophage conditioned media enhance expansion. The role of crypt-specific COX2 will be further investigated using crypt and stem cell- specific COX2 knock-outs. In parallel, the effect of COX2 in macrophages on ISC proliferation and differentiation will be more fully investigated using the enteroid culture system together with bot hypothesis- driven and discovery-driven LC-MS/MS techniques. Finally, Aim 3 will investigate this hypothesis by studying the effect of COX2 including macrophage COX2 on the resolution of inflammation in this model. LC-MS/MS methods for measuring a full panel of pro- inflammatory and inflammation resolving lipid signals will be developed. This LCMS panel will help determine the effect of COX2 on the kinetic balance of pro-inflammatory and inflammation resolving signals within the inflammatory microenvironment of the model. This panel, together with novel lipidomic and proteomic discovery, will also inform the investigation into the effect of macrophage COX2 on ISCs in Aim 2. Given the high morbidity associated with CD, this work will hopefully uncover targets for the amelioration or suppression of this chronic inflammatory disorder.

 描述（由适用提供）：COX2是关于克罗恩病（CD）的遗传敏感性基因座，但是COX2和CD之间的联系的性质尚不清楚。当含有巧克力酵母的饮食挑战时，缺乏COX2的小鼠会出现克罗恩病的炎症。该项目将研究该模型中病理背后的机制，以了解CD的病因。试点数据暗示了COX2在肠上皮屏障完整性和该模型中感染的分辨率中都暗示了COX2，因此COX2缺乏症似乎抑制了上皮组织修复，同时还延长了炎症反应。因此，假设在没有COX2的情况下，屏障完整性改变和长时间感染之间会形成正反馈环，每种反馈循环循环。 AIM 1旨在表征Chate和Cox2对屏障完整性的直接影响。时间过程实验，体外研究和细菌消融都将全部用于隔离巧克力对屏障完整性的直接影响与感染的混杂作用。屏障完整性将通过使用体内易位测定以及测量USSSing室内的跨上皮电阻的测量来部分表征。 AIM 2通过研究COX2对原代肠托动物培养系统内COX2对肠道干细胞（ISC）增殖和分化的影响进一步研究了这一假设。初步数据表明，隐窝内的COX2抑制限制了隐窝的扩展，而外源性PGE2和巨噬细胞的培养基增强扩展。隐窝特异性COX2的作用将使用隐窝和干细胞特异性COX2敲除进一步研究。同时，COX2在巨噬细胞中对ISC增殖的影响和分化将通过肠托体培养系统以及Bot假设驱动和发现驱动的LC-MS/MS技术进行更深入的研究。最后，AIM 3将通过研究包括巨噬细胞COX2对该模型中感染分辨率的COX2的影响来研究这一假设。将开发用于测量全面板促炎和感染脂质信号的LC-MS/MS方法。该LCMS面板将有助于确定COX2对模型炎症微环境中促炎和炎症解析信号的动力学平衡的影响。该面板以及新型的脂肪组和蛋白质组学发现，还将为AIM 2中巨噬细胞COX2对ISC的影响提供投资。鉴于与CD相关的高发病率，这项工作将有望发现对这种慢性炎症性疾病的改善或抑制的目标。