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: 7650889
• 负责人: THOMAS Y MA
• 金额: $ 36万
• 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/7650889
• 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; Gene Proteins; Genes; In Vitro; Inflammation; Inflammation Process; Inflammatory; Inflammatory disease of the intestine; Interleukin-1; Interleukin-1 alpha; Interleukin-1 beta; Interleukins; Intestines; Laboratories; Lead; Left; Mediating; Messenger RNA; MicroRNAs; Mitogen-Activated Protein Kinase 3; 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; public health relevance; 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（由申请人提供）：克罗恩病（CD）患者的肠道紧密连接（TJ）屏障有缺陷，表现为肠道旁细胞通透性增加。肠道 TJ 屏障缺陷是 CD 的重要致病因素，可增加有毒管腔抗原的细胞旁渗透，导致肠道炎症。白介素-1(IL-1()是一种典型的多功能促炎细胞因子，已被证明在胃肠道的肠道炎症过程中发挥核心作用。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道TJ屏障的功能性开放，导致细胞旁细胞增多) 渗透性。本拨款申请的主要目标是阐明介导 IL-1( 诱导肠道 TJ 通透性增加) 的细胞和分子机制，并确定潜在的治疗靶点，以防止 IL-1( 诱导肠道 TJ 通透性增加和随后肠道炎症的发展)，使用体外（由过滤器生长的 Caco-2 肠上皮细胞组成） 单层细胞）和新开发的体内小鼠模型系统。破译IL-1（肠道TJ屏障功能调节）涉及的细胞内途径和分子机制对于推进炎症期间肠道TJ屏障功能障碍的新科学概念以及开发针对TJ屏障预防肠道炎症的新治疗策略具有巨大的潜在科学和临床意义。 初步研究表明，IL-1 诱导的肠上皮 TJ 通透性增加部分是由以下因素介导的：1）细胞外信号调节激酶 1 和 2 (ERK1/2) 信号级联的激活； 2）肌球蛋白轻链激酶（MLCK）基因的激活； 3) microRNA (miRNA) 诱导 occludin mRNA 降解。基于我们的 根据初步数据，我们假设 IL-1（诱导的肠道 TJ 通透性增加部分受到 ERK1/2 信号级联诱导的 MLCK 基因激活和随后 MLCK 蛋白表达的增加以及 microRNA 诱导的 occludin mRNA 降解和 occludin 蛋白消耗的调节。本次拨款申请的具体目标是：1） 描述细胞内信号级联和介导IL-1（诱导肠道TJ通透性增加）的分子过程；2）描述介导IL-1（诱导occludin基因和蛋白表达下调）的细胞和分子机制；3）描述介导IL-1（诱导肠道通透性增加）的机制 体内肠道 TJ 屏障功能的保护和治疗意义。公共卫生相关性：克罗恩病患者患有肠漏症，其特点是肠道对肠腔中有害抗原的通透性增加。本拨款申请中提出的研究旨在确定导致克罗恩病肠漏的细胞内和分子过程。 Interleukin-1( 已被证明在促进克罗恩病和其他肠道炎症性疾病的肠道炎症中发挥重要作用。这些研究的重点将是描绘介导 interleukin-1( 诱导肠道屏障功能紊乱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，以诱导治疗重新收紧或正常化 克罗恩病中的肠漏。