Understanding the importance of IkB-b as a selective co-activator of NF-kB signaling

了解 IkB-b 作为 NF-kB 信号传导选择性共激活剂的重要性

• 批准号: 10153691
• 负责人: Sankar Ghosh
• 金额: $ 40万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10153691
• 关键词: Activator Appliances; Acute; Adaptive Immune System; Address; Amino Acids; Antibodies; Bacterial Infections; Binding; Biochemical; Biological Process; C-terminal; Cell physiology; Cells; ChIP-seq; Chemicals; Chronic; Clinical; Collagen Arthritis; Complement; Complex; Cytoplasm; DNA Binding; DNA analysis; Data; Data Set; Development; Embryonic Development; Enzyme-Linked Immunosorbent Assay; Event; Exposure to; Family; Genes; Genetic Transcription; Genomics; Goals; Infection; Inflammation; Interleukin-6; Knock-in Mouse; Knockout Mice; Knowledge; Laboratories; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mechanics; Mediating; Modeling; Molecular; Mus; Mutant Strains Mice; Mutation; NF-kappa B; Nuclear; Pathogenicity; Pathologic; Pathway interactions; Peptides; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Protein Isoforms; Proteins; Rheumatoid Arthritis; Role; Salmonella typhimurium; Sepsis; Septic Shock; Shapes; Signal Transduction; Stimulus; TNF gene; Techniques; Therapeutic Intervention; Toxic effect; Transcriptional Activation; base; comparative; cytokine; design; dimer; experimental study; in vivo; inhibitor/antagonist; macrophage; mouse model; mutant; novel; overexpression; pathogen; preference; prevent; programs; reconstitution; response; targeted treatment; therapeutic target; transcriptome; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT The transcription factors of the NF-kB family are central regulators of a wide variety of biological processes. This includes events as fundamental as embryonic development and cellular survival as well as the intricate responses of the innate and adaptive immune system. What has remained elusive are the mechanisms that allow this limited set of five proteins to orchestrate the radically different transcriptional programs which are required for these disparate cellular functions. Understanding the details of NF-kB-dependent transcription, however, is of paramount importance for efforts to achieve targeted therapeutic intervention in cases of pathological NF-kB activation, such as sepsis, rheumatoid arthritis and cancer. One of the factors that fine-tune NF-kB activation is the inhibitor of NF-kB, isoform b (IkB-b), which has been shown to serve an intriguing dual role as a conventional inhibitor, and atypical co-activator, of NF-kB- dependent transcription. In unstimulated cells, IkB-b sequesters NF-kB dimers in the cytoplasm and thus prevents transcriptional activation, whereas upon stimulation a nuclear form of IkB-b enhances transcription of a subset of NF-kB-dependent genes. Consequently, IkB-b-deficient macrophages produce starkly reduced levels of the cytokine TNF-a following exposure to an activating stimulus, whereas levels of the cytokine IL-6 are largely unaffected. The goal of this project is to comprehensively understand the role of IkB-b as a selective co-activator of NF-kB activation. This will be accomplished by rigorously characterizing the mechanism that underlies IkB-b function on a biochemical and molecular level. Furthermore, the full extent of IkB-b co-activator function and its effect on global transcription will be probed using cutting-edge genomics techniques, including RNA-seq and ChIP- seq. These data sets will then be integrated to create a holistic model of IkB-b function. Finally, the true physiological relevance of IkB-b will be determined in vivo with mouse models of acute and chronic inflammation, as well as bacterial infection.

项目概要/摘要 NF-kB 家族的转录因子是多种生物过程的核心调节因子。 这包括胚胎发育和细胞存活等基本事件以及复杂的事件 先天性和适应性免疫系统的反应。仍然难以捉摸的是其机制 允许这有限的五种蛋白质来协调完全不同的转录程序，这些程序是 这些不同的细胞功能所需的。了解 NF-kB 依赖性转录的细节， 然而，对于在以下情况下实现有针对性的治疗干预至关重要 病理性 NF-kB 激活，例如败血症、类风湿性关节炎和癌症。 微调 NF-kB 激活的因素之一是 NF-kB 同工型 b (IkB-b) 抑制剂，该抑制剂已被 显示出作为 NF-kB- 的常规抑制剂和非典型共激活剂的有趣双重作用 依赖性转录。在未刺激的细胞中，IkB-b 将 NF-kB 二聚体隔离在细胞质中，从而 阻止转录激活，而在刺激后，IkB-b 的核形式会增强转录 NF-kB 依赖性基因的一个子集。因此，IkB-b 缺陷型巨噬细胞产生的巨噬细胞明显减少。 暴露于激活刺激后细胞因子 TNF-a 的水平，而细胞因子 IL-6 的水平 基本上不受影响。 该项目的目标是全面了解 IkB-b 作为 NF-kB 选择性共激活剂的作用 激活。这将通过严格表征 IkB-b 功能背后的机制来实现 在生化和分子水平上。此外，IkB-b 共激活剂功能的全部范围及其效果 将使用尖端基因组学技术（包括 RNA-seq 和 ChIP）来探究全局转录的情况 序列。然后，这些数据集将被整合以创建 IkB-b 功能的整体模型。最后，真实的 IkB-b 的生理相关性将通过急性和慢性小鼠模型体内确定 炎症，还有细菌感染。