NF-kB /Beta-catenin interaction during intestinal inflammation

肠道炎症期间 NF-kB /β-catenin 相互作用

• 批准号: 7906796
• 负责人: Jun Sun
• 金额: $ 13万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7906796
• 关键词: Bacteria; Bacterial Proteins; Binding; Colitis; Cultured Cells; Cyclin D1; Data; Disease; Enterobacteriaceae; Environment; Epithelial Cells; Genes; Germ-Free; Health; IL8 gene; In Vitro; Inflammation; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Interleukin-10; Intestinal Mucosa; Intestines; Knowledge; Language; Maintenance; Modeling; Mono-S; Mus; NF-kappa B; Pathogenesis; Pathway interactions; Patients; Phosphorylation; Play; Property; Proteins; Research; Role; Signal Pathway; Ubiquitination; beta catenin; c-myc Genes; commensal microbes; in vivo; insight; microbial; novel strategies; response

DESCRIPTION (provided by applicant): Increasing evidence indicates that endogenous enteric bacteria play a crucial role in the pathogenesis of inflammatory bowel diseases (IBDs). It is clear that gut flora contribute to the elevated levels of activated NF-???in the intestinal mucosa of IBD patients. However, it is unclear which bacterial strain and/or bacterial ?products contribute to the pathogenesis and/or maintenance of IBD. Due to the complexity of the gut flora, identification of the specific causative microbial agents in IBD remains challenging, and the search for the causative microbial agents is intense. AvrA is a pathogenic gene of certain bacteria whose encoded protein is inserted into intestinal epithelial cells through a Type III secretory pathway. In previous studies, we demonstrated that AvrA stabilizes ?-catenin, which is a negative regulator of the NF-?? pathway in epithelial cells. We hypothesize that the bacterial effector AvrA inhibits the NF-???pathway and actives the ?-catenin pathway to inhibit intestinal inflammation. In this study, I will focus on the mechanism and effects of AvrA, using both in vitro and in vivo approaches. For in vitro studies, we will colonize epithelial cells with AvrA-sufficient or -deficient bacteria strains. We will determine how the bacterial effector AvrA actually regulates NF-??/?-catenin binding and identify post-transcriptional changes in regulators of the NF-?? and ?-catenin pathways induced by AvrA (e.g., phosphorylations, ubiquitinations). We will investigate expression changes induced by AvrA in downstream targets (i.e., IL-8, c-myc, cyclin D1) of the NF-?? and???-catenin pathways. For in vivo studies, we will employ IL-10-/- mice that develop spontaneous colitis, which has been shown to require colonic bacterial colonization for full expression. These IL-10-/- mice will initially be raised in germ-free environments, and will then be mono-associated with E.coli F18 or E.coli F18AvrA+ to determine the function of AvrA in inhibiting inflammation in vivo. Elucidating the mechanisms of AvrA regulating the NF-?? and ?-catenin signaling pathways will provide new insights into how bacteria-host interactions contribute to inflammation. These studies have direct relevance to a better understanding of diseases such as inflammatory bowel diseases and infectious colitis. Lay language: This proposal is aimed at understanding the mechanism and effects of the bacterial protein AvrA in inhibiting inflammation. We will focus on the mechanism and effects of AvrA in cultured cells colonized with bacterial strains (with or without AvrA expression) and in a mouse colitis model. The information obtained from our research will provide new insights into the way bacteria-host interactions contribute to inflammation. This has direct relevance to a better understanding of diseases such as inflammatory bowel diseases and infectious colitis.

描述（由申请人提供）：越来越多的证据表明，内源性肠道细菌在炎症性肠病（IBD）的发病机制中起着至关重要的作用。很明显，肠道植物群有助于激活NF-κ B水平的升高。在IBD患者的肠粘膜中。然而，目前还不清楚是哪种细菌菌株和/或细菌？产品有助于IBD的发病机制和/或维持。由于肠道植物群的复杂性，IBD中特定致病微生物的鉴定仍然具有挑战性，并且对致病微生物的寻找是激烈的。AvrA是某些细菌的致病基因，其编码的蛋白质通过III型分泌途径插入肠上皮细胞。在以前的研究中，我们证明了AvrA稳定？-连环蛋白是NF-κ B的负调节因子。pathway途径in epithelial上皮cell细胞.我们假设细菌效应子AvrA抑制NF-κ B的表达。途径和活动？catenin途径抑制肠道炎症。在这项研究中，我将集中在AvrA的机制和影响，在体外和体内的方法。对于体外研究，我们将用AvrA充足或缺乏的细菌菌株定殖上皮细胞。我们将确定细菌效应子AvrA实际上是如何调节NF-κ B/κ B的。连环蛋白结合，并确定转录后的变化，调节NF-？？然后呢？AvrA诱导的连环蛋白途径（例如，磷酸化、泛素化）。我们将研究AvrA在下游靶点（即，IL-8、c-myc、cyclin D1）的表达。然后呢？连环蛋白通路对于体内研究，我们将采用发展自发性结肠炎的IL-10-/-小鼠，其已显示需要结肠细菌定植以完全表达。这些IL-10-/-小鼠最初将在无菌环境中饲养，然后将与大肠杆菌F18或大肠杆菌F18 AvrA+单相关，以确定AvrA在体内抑制炎症中的功能。阐明AvrA调节NF-κ B的机制。然后呢？连环蛋白信号通路将为细菌-宿主相互作用如何促进炎症提供新的见解。这些研究与更好地理解炎症性肠病和感染性结肠炎等疾病直接相关。 外行语言：该提案旨在了解细菌蛋白AvrA抑制炎症的机制和作用。我们将专注于AvrA在细菌菌株（有或没有AvrA表达）定植的培养细胞和小鼠结肠炎模型中的机制和作用。从我们的研究中获得的信息将为细菌-宿主相互作用促进炎症的方式提供新的见解。这与更好地理解炎症性肠病和感染性结肠炎等疾病有直接关系。