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: 10404063
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10404063
• 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; 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.

项目摘要/摘要 NF-KB家族的转录因子是各种生物学过程的中心调节剂。 这包括像胚胎发育和细胞生存一样基本的事件以及复杂的事件 先天和适应性免疫系统的反应。仍然难以捉摸的是机制 允许这套有限的五种蛋白质来协调截然不同的转录程序 这些不同的细胞功能所需。了解NF-KB依赖性转录的细节， 但是，对于在为实现有针对性的治疗干预措施的情况下， 病理NF-KB激活，例如败血症，类风湿关节炎和癌症。 微调NF-KB激活的因素之一是NF-KB的抑制剂，同工型B（IKB-B），它一直是 作为常规抑制剂和非典型共激活因子，表现出有趣的双重作用，NF-KB- 依赖的转录。在未刺激的细胞中，IKB-B隔离会在细胞质中的NF-KB二聚体，因此 防止转录激活，而刺激IKB-B的核形式会增强 NF-KB依赖性基因的子集。因此，IKB-B缺陷巨噬细胞产生明显的降低 暴露于激活刺激后的细胞因子TNF-A水平，而细胞因子IL-6的水平 在很大程度上不受影响。 该项目的目的是全面了解IKB-B作为NF-KB的选择性共激活器的作用 激活。这将通过严格表征IKB-B功能的机制来实现 在生化和分子水平上。此外，IKB-B共激活因子及其效果的全部范围 在全球转录上将使用尖端的基因组技术进行探测，包括RNA-Seq和ChIP- seq。然后将集成这些数据集以创建IKB-B函数的整体模型。最后，真实 IKB-B的生理相关性将用急性和慢性小鼠模型在体内确定 炎症以及细菌感染。