Mechanistically defining the role for each NFkB family member in pro-inflammatory macrophage activation

从机制上定义每个 NFkB 家族成员在促炎巨噬细胞激活中的作用

• 批准号: 10527312
• 负责人: Allison E Daly
• 金额: $ 3.74万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10527312
• 关键词: 2019-nCoV; Acute Respiratory Distress Syndrome; Affinity; Bacterial Infections; Binding; Binding Sites; Biochemical; Biological; Biological Models; Biological Process; Bone Marrow; COVID-19 patient; CRISPR/Cas technology; Cause of Death; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; ChIP-seq; Chromatin; Clinical; Combinatorics; Complex; Country; Coupled; Data; Data Set; Dependence; Deterioration; Developing Countries; Disease; Escherichia coli; Event; Family; Family member; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Homo; Immune; Immune System Diseases; Immune system; In Vitro; Income; Individual; Infection; Inflammatory; LCN2 gene; Lead; Link; Logic; Macrophage Activation; Measures; Modeling; Molecular; Mutate; Nuclear; Organ; Pathogenesis; Pathogenicity; Pathology; Patients; Pattern recognition receptor; Peritoneal Macrophages; Persons; Phenotype; Pregnancy; Proteins; Regulation; Reproducibility; Research; Research Proposals; Role; Sepsis; Signal Transduction; Stimulus; System; TNFRSF5 gene; Transactivation; Viral; Virus; Virus Diseases; World Health Organization; cecal ligation puncture; cell behavior; chemokine; combinatorial; cytokine; dimer; genetic regulatory protein; genome-wide; immune activation; immunoregulation; in vivo; insight; macrophage; member; monomer; mortality; mouse model; novel; patient population; programs; response; sepsis induced ARDS; systemic inflammatory response; therapeutic target; transcription factor; transcriptome; transcriptome sequencing

PROJECT SUMMARY The World Health Organization (WHO) estimates that ~6 million people die from sepsis each year, with the greatest burden felt in low to middle income countries. According to the WHO, one out of ten deaths during pregnancy is a result of maternal sepsis with over 95% of these cases arising in developing countries. Finally, the Centers for Disease Control and Prevention, estimates that 3-17% of all patients with COVID-19 will develop acute respiratory distress syndrome (ARDS), the primary cause of mortality in this patient population. Sepsis and ARDS, represent severe consequences of immune system overactivation and dysregulation during infection. Although the burden of this pathology is recognized, the molecular underpinnings of this clinical deterioration remain poorly understood. NFkB is a ubiquitous transcription factor that is activated during viral and bacterial infection to control innate immune cell behavior and is thus likely to be a key player in immune dysregulation. NFkB is a family of TFs containing 5 subunits (cRel, RelA, p50, p52, and RelB) that combinatorically interact to form 15 possible dimeric species to regulate gene expression. Although, NFkB is well studied, there remains a paucity of research investigating the function of individual NFkB dimers in immune cell activation. Preliminary analysis supports the central hypothesis that each dimer of NFkB has a unique biological function. Understanding the role of individual NFkB dimers could open avenues for the precise modulation of pro-inflammatory gene expression. Further, my data supports the hypothesis that cRel homodimers are necessary for the selective regulation of a small group of genes including Il12b; and p50 homodimers associate with a co-regulatory protein to activate key pro-inflammatory genes including Il6, Il1b and Lcn2. This research employs a bone marrow derived macrophage model system in combination with the extraction of primary peritoneal macrophages stimulated ex vivo with E. coli or in vivo with a cecal ligation and puncture model of sepsis, to define dimer specific roles of NFkB in macrophage activation and the molecular mechanisms underlying dimer specific gene regulation. This proposal will i) explore the mechanistic role of cRel homodimers in macrophage activation; and ii) investigate p50 homodimer specific gene regulation in macrophage activation. This research proposal focuses on cRel and p50 homodimers due to their potent activation downstream of viral and bacterial PRRs and their highly specific roles in immune cell activation. Upon completion, this research will provide novel insights into the regulatory logic employed by NFkB by revealing the underlying mechanisms that make dimer specific functions possible. Further, it could uncover specific therapeutic targets for immune system modulation to help slow or halt exaggerated immune cell activation seen in sepsis and ARDS.

项目摘要 世界卫生组织（WHO）估计每年约有600万人死于败血症， 低至中收入国家的最大负担。根据世卫组织的说法，十分之二的死亡 怀孕是孕产妇败血症的结果，其中95％的病例在发展中国家出现。最后， 疾病控制和预防中心估计所有Covid-19患者中有3-17％将发展 急性呼吸窘迫综合征（ARDS），这是该患者人群死亡率的主要原因。败血症 和ARDS，代表了免疫系统过度激活和感染期间失调的严重后果。 尽管确认了这种病理的负担，但该临床恶化的分子基础 保持不当理解。 NFKB是一种无处不在的转录因子，在病毒和细菌期间被激活 控制先天免疫细胞行为的感染，因此很可能是免疫失调的关键参与者。 NFKB是一个包含5个亚基（CREL，RELA，P50，P52和RELB）的TFS家族，它们合并与 表格15种可能的二聚体物种调节基因表达。虽然，对NFKB进行了充分的研究，但仍有一个 研究研究单个NFKB二聚体在免疫细胞激活中的功能的研究。初步的 分析支持一个中心假设，即NFKB的每个二聚体具有独特的生物学功能。理解 单个NFKB二聚体的作用可以为促炎基因的精确调节开放途径 表达。此外，我的数据支持以下假设，即CREL同型二聚体对于选择性是必要的 调节包括IL12B在内的一小部分基因；和P50同型二聚体与共调控蛋白相关 激活包括IL6，IL1B和LCN2在内的关键促炎基因。该研究采用骨髓 衍生的巨噬细胞模型系统与原发性腹膜巨噬细胞的提取结合 用大肠杆菌或体内刺激败血症的盲肠和穿刺模型，以定义二聚体 NFKB在巨噬细胞激活和二聚体特异性基因的分子机制中的特定作用 规定。该提议将i）探讨Crel同二聚体在巨噬细胞激活中的机械作用；和 ii）研究巨噬细胞激活中p50均二聚体特异性基因调节。这项研究建议重点 在CREL和P50同二聚体上，由于其在病毒和细菌PRR下游的有效激活及其 在免疫细胞激活中高度特异性的作用。完成后，这项研究将为您提供新的见解 NFKB采用的调节逻辑是通过揭示使二聚体特定功能的基本机制 可能的。此外，它可以发现免疫系统调制的特定治疗靶标，以帮助慢速或停止 败血症和ARDS中看到的夸张的免疫细胞激活。