Deciphering the role of Claudin-23 in regulation of the intestinal epithelial barrier

解读 Claudin-23 在肠上皮屏障调节中的作用

• 批准号: 10646214
• 负责人: Kristen Michelle Lozada-Soto
• 金额: $ 4.28万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10646214
• 关键词: Actins; Active Sites; Adherent Culture; Antigens; Apical; Binding Sites; Biochemical; Biological Assay; Biopsy; Cell Line; Cell membrane; Cell physiology; Cells; Cellular biology; Chronic; Colon; Complement; Complex; Cytoplasm; Cytoskeleton; DNA Binding; Data; Development; Digestive System Disorders; Disease; Down-Regulation; Economic Burden; Environmental Risk Factor; Epithelial Cells; Epithelium; Etiology; Event; Family; Family member; Functional disorder; Gastrointestinal Diseases; Genetic Predisposition to Disease; Genetic Transcription; Goals; HIF1A gene; Health; Healthcare Systems; Homeostasis; Human; Hypoxia; Hypoxia Inducible Factor; Immune; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Integral Membrane Protein; Intercellular Junctions; Intestines; Investigation; Knock-out; Knockout Mice; Knowledge; Lateral; Link; Membrane; Messenger RNA; Modeling; Molecular; Mucous Membrane; Mus; Nutrient; Oral; Pathologic; Patients; Permeability; Physicians; Physiological; Positioning Attribute; Property; Proteins; Quality of life; Reagent; Regulation; Role; Scaffolding Protein; Scientist; Signal Transduction; Spatial Distribution; Specificity; Stimulus; Techniques; Therapeutic Intervention; Tight Junctions; Tissues; Training; base; career; chromatin immunoprecipitation; colonic crypt; differential expression; disease mechanisms study; fluorescein isothiocyanate dextran; gastrointestinal symptom; gut homeostasis; in silico; in vivo; intestinal barrier; intestinal crypt; intestinal epithelium; knock-down; mRNA Expression; member; monolayer; murine colitis; normoxia; novel; novel therapeutic intervention; overexpression; promoter; protein expression; seal; tool; transcription factor; uptake; villin

PROJECT SUMMARY Intestinal epithelial cells (IECs) constitute a selective barrier that regulates nutrient uptake and restricts antigen access from the lumen to the underlying mucosa. Permeability across the intestinal epithelium is dynamically regulated by a range of physiologic and pathologic stimuli. Patients with chronic inflammatory bowel disease (IBD), suffer from debilitating gastrointestinal symptoms that significantly impact their quality of life and pose a disproportionately large economic burden on the healthcare system. Although genetic susceptibility, environmental factors, and immune dysregulation are all critical contributors to the multifactorial etiology of IBD, there is increasing evidence suggesting a central role for intestinal epithelial barrier dysfunction in IBD patients. Intestinal barrier properties are regulated through dynamic remodeling and maturation of intercellular junctions along the lateral membrane of IECs. Tight junctions (TJs) are the most apical intercellular junctional complex and are composed of a highly organized array of transmembrane proteins and cytoplasmic scaffolding proteins that anchor the complex to the actin cytoskeleton. The claudin (CLDN) family of TJ transmembrane proteins is comprised of 27 members in humans that control barrier function by regulating the sealing properties of the TJ. In the colon, CLDNs are differentially expressed to spatially regulate barrier properties. Notably, CLDN expression is perturbed in IBD and has been linked to barrier compromise. However, the mechanisms by which IECs differentially express CLDNs to control barrier function and how CLDN dysregulation contributes to epithelial barrier compromise in IBD remain unclear. In this proposal we present compelling preliminary data implicating CLDN23, an understudied non-classical claudin family member, as a central orchestrator of CLDN complex stability and TJ barrier function in IECs. Moreover, we have observed that CLDN23 expression is dysregulated in IBD. Thus, my proposed studies will employ novel tools to investigate how CLDN23 controls intestinal epithelial barrier function using complementary in vitro and in vivo techniques. Successful completion of the proposed studies will expand our knowledge of basic mechanisms that control intestinal epithelial barrier function in health and disease, and will open new avenues for investigation of therapeutic interventions to restore barrier properties and ameliorate inflammation in IBD. Importantly, this scientific proposal and training will position me for a sustained career as a physician-scientist studying the mechanisms of disease of gastrointestinal inflammatory illnesses.

项目摘要 肠上皮细胞（IEC）构成了调节营养吸收并限制的选择性屏障 抗原从管腔进入基础粘膜。横跨肠上皮的渗透性为 由一系列生理和病理刺激动态调节。慢性炎症患者 疾病（IBD），胃肠道症状令人衰弱，严重影响其生活质量和 在医疗保健系统上造成不成比例的经济负担。虽然遗传敏感性，但 环境因素和免疫失调都是IBD多因素病因的关键因素， 越来越多的证据表明，IBD患者肠上皮屏障功能障碍的核心作用。 通过动态重塑和细胞间连接的成熟来调节肠道屏障特性 沿着IEC的外侧膜。紧密连接（TJ）是最根尖的细胞间连接络合物 由高度组织的跨膜蛋白和细胞质脚手架蛋白组成 将复合物锚定在肌动蛋白细胞骨架上。 TJ跨膜蛋白的Claudin（CLDN）家族是 由人类中的27名成员组成，通过调节TJ的密封特性来控制屏障功能。 在结肠中，CLDN差异表达为空间调节屏障特性。值得注意的是，CLDN 表达在IBD中受到干扰，并与障碍妥协有关。但是， IEC差异表达CLDN以控制屏障功能以及CLDN失调如何促进 IBD中的上皮屏障妥协尚不清楚。在此提案中，我们提出引人注目的初步数据 暗示了一个研究的非古典克劳丁家族成员的Cldn23，是CLDN的中央编排者 IEC中的复杂稳定性和TJ屏障功能。而且，我们已经观察到Cldn23的表达是 IBD失调。因此，我提出的研究将采用新颖的工具来研究CLDN23如何控制 肠上皮屏障功能使用互补的体外和体内技术。成功完成 拟议的研究将扩大我们对控制肠上皮屏障的基本机制的了解 在健康和疾病中发挥作用，并将为调查治疗干预措施开辟新的途径 IBD中的屏障特性和改善炎症。重要的是，这个科学的建议和培训将 将我定位为医生科学家的持续职业，研究疾病的机制 胃肠道炎症性疾病。