Interleukin-1 Beta Modulation of Intestinal Tight Junction Barrier

Interleukin-1 Beta 调节肠道紧密连接屏障

• 批准号: 8098031
• 负责人: THOMAS Y MA
• 金额: $ 31.98万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8098031
• 关键词: Antigens; Applications Grants; Biological Models; Biological Preservation; Crohn' s disease; Data; Defect; Development; Disease; Down-Regulation; Epithelial; Epithelial Cells; Functional disorder; Gastrointestinal tract structure; Gene Expression; Genes; Health; In Vitro; Inflammation; Inflammation Process; Inflammatory; Inflammatory disease of the intestine; Interleukin-1; Interleukin-1 beta; Interleukins; Intestines; Laboratories; Lead; Left; Mediating; Messenger RNA; MicroRNAs; Mitogen-Activated Protein Kinases; Molecular; Mus; Myosin Light Chain Kinase; Nuclear; Pathway interactions; Patients; Perfusion; Permeability; Play; Process; Proteins; Regulation; Role; Side; Signal Pathway; Signal Transduction; Suggestion; Testing; Therapeutic; Therapeutic Agents; Tight Junctions; Time; Treatment Efficacy; Up-Regulation; base; clinically relevant; cytokine; in vivo; in vivo Model; mRNA Expression; monolayer; mouse model; novel; novel therapeutics; occludin; prevent; protein expression; response; therapeutic target; transcription factor

DESCRIPTION (provided by applicant): Patients with Crohn's disease (CD) have a defective intestinal tight junction (TJ) barrier, manifested by an increase in intestinal paracellular permeability. The defective intestinal TJ barrier is an important pathogenic factor of CD that allows increased paracellular permeation of toxic luminal antigens, leading to intestinal inflammation. Interleukin-1( (IL-1(), a prototypical multi-functional pro-inflammatory cytokine, has been shown to play a central role in the intestinal inflammation process in the gastrointestinal tract. Recent studies indicate that an important pro-inflammatory action of IL-1( and other pro-inflammatory cytokines is to cause a functional opening of the intestinal TJ barrier, leading to an increase in paracellular permeability. The broad objectives of this grant application are to elucidate the cellular and molecular mechanisms that mediate the IL-1( induced increase in intestinal TJ permeability and to identify potential therapeutic targets to prevent the IL-1( induced increase in intestinal TJ permeability and subsequent development of intestinal inflammation, using an in-vitro (consisting of filter-grown Caco-2 intestinal epithelial monolayers) and a newly developed in-vivo mouse model system. Deciphering the intracellular pathways and the molecular mechanisms involved in IL-1( modulation of intestinal TJ barrier function has great potential scientific and clinical relevance in advancing novel scientific concepts regarding intestinal TJ barrier dysfunction during inflammation and for developing new therapeutic strategies that target the TJ barrier in preventing intestinal inflammation. Our preliminary studies suggested that the IL-1( induced increase in intestinal epithelial TJ permeability was mediated in part by: 1) activation of extracellular signal- regulated kinases 1 and 2 (ERK1/2) signaling cascade; 2) activation of myosin light chain kinase (MLCK) gene; and 3) microRNA (miRNA) induced degradation of occludin mRNA. Based on our preliminary data, we hypothesize that the IL-1( induced increase in intestinal TJ permeability is regulated in part by ERK1/2 signaling cascade induced activation of MLCK gene and subsequent increase in MLCK protein expression and by microRNA induced degradation of occludin mRNA and depletion of occludin protein. The proposed specific aims of this grant application are to: 1) delineate the intracellular signaling cascade and the molecular processes that mediate the IL-1( induced increase in intestinal TJ permeability; 2) delineate the cellular and molecular mechanisms that mediate the IL-1( induced down-regulation of occludin gene and protein expression; and 3) delineate the mechanisms that mediate the IL-1( induced increase in intestinal permeability in-vivo and the therapeutic implications of preservation of intestinal TJ barrier function in-vivo. PUBLIC HEALTH RELEVANCE: Patients with Crohn's disease have a leaky gut, characterized by an increase in intestinal permeability to harmful antigens in the intestinal lumen. The studies proposed in this grant application seek to determine the intracellular and molecular processes that contribute to the leaky gut of Crohn's disease. Interleukin-1( has been shown to play an important role in promoting intestinal inflammation in Crohn's disease and other inflammatory conditions of the gut. The focus of these studies will be to delineate the intracellular and molecular processes that mediate the interleukin-1( induced disturbance of intestinal barrier function. The proposed studies also seek to identify potential therapeutic targets and strategies to induce therapeutic re-tightening or normalization of leaky gut in Crohn's disease.

描述（由申请方提供）：克罗恩病（CD）患者存在肠紧密连接（TJ）屏障缺陷，表现为肠细胞旁通透性增加。缺陷的肠TJ屏障是CD的重要致病因素，其允许毒性管腔抗原的细胞旁渗透增加，导致肠道炎症。白细胞介素-1（IL-1）是一种典型的多功能促炎细胞因子，在胃肠道炎症过程中起着重要作用。最近的研究表明，IL-1（和其他促炎细胞因子）的重要促炎作用是引起肠TJ屏障的功能性开放，导致细胞旁通透性增加。这项资助申请的主要目的是阐明介导IL-1的细胞和分子机制。（诱导的肠TJ渗透性增加）和鉴定潜在的治疗靶点以预防IL-1（诱导的肠TJ渗透性增加和随后的肠道炎症发展，使用体外（由过滤生长的Caco-2肠上皮单层组成）和新开发的体内小鼠模型系统。解读参与IL-1（肠TJ屏障功能的调节）的细胞内途径和分子机制在推进关于炎症期间肠TJ屏障功能障碍的新科学概念和开发靶向TJ屏障以预防肠道炎症的新治疗策略方面具有巨大的潜在科学和临床相关性。我们的初步研究表明，IL-1诱导的肠上皮TJ渗透性的增加部分由以下介导：1）细胞外信号调节激酶1和2（ERK 1/2）信号级联的激活; 2）肌球蛋白轻链激酶（MLCK）基因的激活;和3）微小RNA（miRNA）诱导的闭合蛋白mRNA的降解。基于我们的初步数据，我们假设IL-1诱导的肠TJ渗透性的增加部分地由ERK 1/2信号级联诱导的MLCK基因的激活和随后的MLCK蛋白表达的增加以及由microRNA诱导的occludin mRNA的降解和occludin蛋白的消耗来调节。本基金申请的具体目标是：1）描述细胞内信号级联和介导IL-1的分子过程（诱导肠TJ渗透性增加; 2）描绘介导IL-1的细胞和分子机制，（诱导occludin基因和蛋白表达下调;以及3）描述了介导IL-1诱导的体内肠通透性增加的机制以及体内肠TJ屏障功能保留的治疗意义。公共卫生相关性：克罗恩病患者存在肠道渗漏，其特征是肠道对肠腔中有害抗原的渗透性增加。本研究申请旨在确定导致克罗恩病肠漏的细胞内和分子过程。白细胞介素-1（Interleukin-1，IL-1）已被证明在促进克罗恩病（Crohn's disease）的肠道炎症和肠道的其他炎症性病症中起重要作用。这些研究的重点将是描绘介导白细胞介素-1（诱导肠屏障功能障碍）的细胞内和分子过程。拟议的研究还寻求确定潜在的治疗靶点和策略，以诱导克罗恩病中漏肠的治疗性再收紧或正常化。