Mechanisms of epithelial barrier dysfunction mediated by inflammatory cytokines

炎症细胞因子介导的上皮屏障功能障碍机制

• 批准号: 7539581
• 负责人: Christopher Weber
• 金额: $ 5.29万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7539581
• 关键词: Affect; American; Area; Biophysics; Brain; Cardiac; Cell membrane; Cells; Cellular biology; Clinical; Condition; Coupled; Cultured Cells; Data; Defect; Development; Diagnostic; Disease; Disruption; Electric Capacitance; Electrodes; Electrophysiology (science); Environment; Epithelial; Epithelial Cells; Event; Extracellular Space; Functional disorder; Future; Genetic Transcription; Goals; Health; Hour; Human; Immune; Immune system; Individual; Inflammatory; Inflammatory Bowel Diseases; Interleukin-13; Intestinal Diseases; Ions; Kidney; Knowledge; Laboratories; Liver; Lung; Malabsorption Syndromes; Measurement; Measures; Mediating; Medical; Medicine; Membrane; Mentors; Methods; Microscope; Molecular; Morbidity - disease rate; Nutrient; Operative Surgical Procedures; Organ; Pathologist; Pathology; Pathway interactions; Patients; Permeability; Personal Satisfaction; Physiological; Protein Isoforms; Proteins; Qualifying; Regulation; Research; Research Project Grants; Resistance; Resolution; Resources; Signal Pathway; Sodium-Calcium Exchanger; Standards of Weights and Measures; Structure; Surface; TNF gene; Techniques; Testing; Therapeutic; Thinking; Tight Junctions; Time; Time Study; Treatment Protocols; Voltage-Clamp Technics; Work; base; cytokine; day; design; gastrointestinal; improved; inhibitor/antagonist; innovation; insight; intracellular protein transport; millisecond; novel; novel strategies; professor; protein expression; protein transport; seal; skills; tool; voltage clamp

DESCRIPTION (provided by applicant): A single layer of epithelial cells forms the barrier between the harsh environment of the gut lumen and the interstitium. In intestinal diseases such as inflammatory bowel disease, tight junctions, which seal the intercellular space, become leaky and the barrier is compromised. Inflammatory cytokines are known to contribute to this barrier defect, howevertheir mechanisms of action are poorly defined. The long term objectives of this application are to understand the unique regulatory mechanism that different cytokines have on barrier function. My central hypothesis, based on the preliminary data, is that different cytokines affect barrier function via functionally distinct mechanisms. I will test this hypothesis using standard tools as well as a novel approach I have developed that will allow high resolution analysis of tight junction biophysics. The predominant methods used in this study will allow biophysical characterization of the tight junction barrier and the effects of inflammatory cytokines. The first aim will characterize the distinct effectsof different cytokines, specifically TNF and IL13,on barrier function. Methods will include traditional current clamp and ion selectivity measurements coupled with macromolecular flux studies. These functional measurements will be correlated with measurements of protein transcription and expression. The regulatory mechanisms involved will be assessed through pharmacologic manipulation. The second aim will take advantage of and refine the approach I have developed to make high resolution biophysical measurements of barrier dynamics under normal conditions. These techniques will allow measurement of barrier function over extremely small areas on a millisecond time scale. The third aim will combine observations and techniques of the first two aims to determine the distinct effects of different cytokines on the biophysics of tight junction barrier function at high resolution. Completion of these aims will have significant positive effects on human health because it will greatly enhance our understanding of specific mechanisms by which cytokines cause barrier dysfunction. This will allow development of specific therapies targeting distinct pathways of barrier dysfunction. This holds great promise as we enter the era of individualized medicine, since availability of agents targeting different mechanisms of barrier dysfunction will allow treatment regimens designed for individual patients. Additionally, the techniques developed in the second aim will have utility in studying tight junction biophysics in other organs, such as kidney, lung, liver, and brain, where tight junction disruption is thought to contribute to disease.

描述（由申请人提供）：单层上皮细胞形成肠腔恶劣环境和间质之间的屏障。在炎症性肠病等肠道疾病中，密封细胞间隙的紧密连接变得渗漏，屏障受到损害。已知炎症细胞因子会导致这种屏障缺陷，但其作用机制尚不清楚。该应用的长期目标是了解不同细胞因子对屏障功能的独特调节机制。基于初步数据，我的中心假设是不同的细胞因子通过功能不同的机制影响屏障功能。我将使用标准工具以及我开发的新方法来测试这个假设，该方法将允许对紧密连接生物物理学进行高分辨率分析。本研究中使用的主要方法将允许对紧密连接屏障和炎症细胞因子的影响进行生物物理表征。第一个目标是描述不同细胞因子（特别是 TNF 和 IL13）对屏障功能的不同影响。方法将包括传统的电流钳和离子选择性测量以及大分子通量研究。这些功能测量将与蛋白质转录和表达的测量相关。所涉及的调节机制将通过药理学操作进行评估。第二个目标将利用并完善我开发的方法，以在正常条件下对屏障动力学进行高分辨率生物物理测量。这些技术将允许在毫秒时间尺度上测量极小区域的屏障功能。第三个目标将结合前两个目标的观察和技术，以确定不同细胞因子对高分辨率紧密连接屏障功能的生物物理学的不同影响。这些目标的完成将对人类健康产生显着的积极影响，因为它将极大地增强我们对细胞因子导致屏障功能障碍的具体机制的理解。这将有助于开发针对屏障功能障碍的不同途径的特定疗法。随着我们进入个体化医疗时代，这带来了巨大的希望，因为针对不同屏障功能障碍机制的药物的可用性将允许为个体患者设计治疗方案。此外，第二个目标中开发的技术将可用于研究其他器官的紧密连接生物物理学，例如肾、肺、肝脏和大脑，这些器官中的紧密连接破坏被认为会导致疾病。