Glucocorticoid-regulated transcription networks in macrophage biology

巨噬细胞生物学中糖皮质激素调节的转录网络

• 批准号: 10432063
• 负责人: INEZ ROGATSKY
• 金额: $ 49.28万
• 依托单位: HOSPITAL FOR SPECIAL SURGERY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10432063
• 关键词: ATAC-seq; Adipocytes; Adipose tissue; Amputation; Anti-Inflammatory Agents; Binding Sites; Biology; Blood Vessels; Bone Marrow; Cells; Chromatin; Chronic; Complex; Data; Disease; EP300 gene; Enhancers; Environment; Equilibrium; Event; Exposure to; Family; Fatty Acids; Fatty acid glycerol esters; GKLF protein; GRIP1 gene; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Goals; Growth; Health; Heart Diseases; Histopathology; IL4 gene; ITGAX gene; Immune; Impaired wound healing; In Vitro; Individual; Inflammation; Inflammatory; Insulin; Interleukin 4 Receptor; Interleukin-1 beta; Interleukin-4; Lead; Link; Liver; Maintenance; Maps; Mediating; Mediator of activation protein; Metabolic; Metabolic Diseases; Metabolic dysfunction; Molecular; Mus; NCOA2 gene; NRIP1 gene; Non-Insulin-Dependent Diabetes Mellitus; Nuclear; Nuclear Receptor Gene; Nuclear Receptors; Obesity; Outcome; PPAR gamma; Pathogenesis; Pathogenicity; Pathway interactions; Phenotype; Physiological; Population; Property; Proteins; Research; Role; STAT6 gene; Signal Transduction; Skin; Specific qualifier value; Stimulus; Structure; Surface; TNF gene; Testing; Thinness; Transforming Growth Factors; Ursidae Family; Vascular Endothelium; Wound models; blood damage; chemokine; chronic ulcer; cofactor; comorbidity; cytokine; epigenome; epigenomics; fatty acid oxidation; genome-wide; in vivo; macrophage; metabolic profile; monocyte; novel; oxidation; programs; promoter; response; single-cell RNA sequencing; skin damage; tissue repair; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; uptake; virtual; wound; wound healing

Macrophage (MΦ)-driven inflammation is central to the pathogenesis of metabolic disease in adipose tissue, liver, vascular endothelium and the skin. However, MΦ are extremely diverse in their ontogeny, epigenomes, transcriptomes and function. Indeed, `lean' adipose tissue M2-like MΦ are anti-inflammatory, facilitate fatty acid oxidation and sensitize adipocytes to insulin. In the skin, homeostatic tissue-repairing MΦ are essential responders to damage, and defective wound healing is a striking comorbidity of metabolic disorder. Signals for homeostatic MΦ programming (e.g., interleukin [IL]4 and glucocorticoids [GC]) initiate via cognate transcription factors (STAT6 and the GC nuclear receptor [GR]) gene expression cascades that ultimately converge upon the `master regulator' kruppel-like factor (KLF)4 that activates many M2-specific genes. Conversely, M1-like inflammatory MΦ bear the transcriptional signature of nuclear factor (NF)κB and overproduce mediators of chronic inflammation (TNF, IL1β, iNOS, Ccl2). Although an M1/M2 imbalance is strongly linked to metabolic dysfunction and impaired wound healing, the specific epigenomic and transcriptional networks underlying homeostatic polarization of different M2 populations remain obscure. In fact, only limited data exist on the mechanisms of KLF4 function as a transcription factor, and virtually none on its genome-wide distribution or role in programming individual M2 subsets. Unexpectedly, we discovered that a nuclear receptor cofactor – the GR-interacting protein (GRIP)1 – serves as a coactivator for KLF4 facilitating M2 polarization of mouse bone marrow-derived MΦ. Given that Klf4 itself is a GR target, GRIP1 could mediate the multi-level integration of M2 transcription programs in vivo when MΦ encounter distinct polarizing signals, e.g., GC and IL4, simultaneously. The objective of this application is to understand the epigenomics, transcriptomics and higher order chromatin interactions of homeostatic MΦ in vitro and in vivo. Our central hypothesis is that GC and IL4 create partially overlapping yet distinct chromatin environments in homeostatic MΦ, and that by serving as a shared cofactor for GR and KLF4, GRIP1 facilitates the physiologically relevant functional convergence of M2- like transcription programs in vivo. Our Specific Aims are to: 1) Dissect the global contribution of GRIP1 to GR and KLF4 enhancer formation and to the core M2-like transcription program through genome-wide approaches in GC- and IL4-polarized MΦ ex vivo; 2) Assess the impact of GRIP1 loss on the phenotypic and metabolic properties of homeostatic MΦ ex vivo and on their ability to undergo epigenomic and transcriptional programming, and support tissue repair in vivo; 3) Chart the first map of higher-order chromatin and GRIP1- dependent enhancer-promoter interactions in homeostatic MΦ, and identify the mechanistic determinants of the GRIP1:KLF4 cross-talk. The successful completion of this project will yield a comprehensive analysis of global chromatin interactions, the core transcriptome as well as GRIP1-dependent molecular mechanisms of homeostatic programming of MΦ which, unlike inflammatory MΦ activation, remains poorly defined.

巨噬细胞（MΦ）驱动的炎症是脂肪组织代谢疾病发病机制的核心， 肝脏、血管内皮和皮肤。然而，MΦ 在个体发育、表观基因组、 转录组和功能。事实上，“瘦”脂肪组织 M2 样 MΦ 具有抗炎作用，促进脂肪酸生成 氧化并使脂肪细胞对胰岛素敏感。在皮肤中，稳态组织修复 MΦ 至关重要 对损伤的反应和伤口愈合缺陷是代谢紊乱的一个显着的合并症。信号为 稳态 MΦ 编程（例如，白细胞介素 [IL]4 和糖皮质激素 [GC]）通过同源转录启动 因子（STAT6 和 GC 核受体 [GR]）基因表达级联最终汇聚于 激活许多 M2 特异性基因的“主调节器”kruppel 样因子 (KLF)4。相反，类似M1 炎症 MΦ 具有核因子 (NF)κB 的转录特征，并过度产生以下介质： 慢性炎症（TNF、IL1β、iNOS、Ccl2）。尽管 M1/M2 失衡与代谢密切相关 功能障碍和伤口愈合受损，特定的表观基因组和转录网络 不同 M2 群体的稳态极化仍然不清楚。事实上，关于这一点的数据非常有限。 KLF4 作为转录因子发挥作用的机制，实际上对其全基因组分布或 在对各个 M2 子集进行编程时的作用。出乎意料的是，我们发现了一个核受体辅助因子—— GR 相互作用蛋白 (GRIP)1 – 作为 KLF4 的共激活剂，促进小鼠骨的 M2 极化 骨髓来源的 MΦ。鉴于Klf4本身是GR靶标，GRIP1可以介导M2的多级整合 当 MΦ 同时遇到不同的极化信号（例如 GC 和 IL4）时，体内转录程序。 该应用程序的目的是了解表观基因组学、转录组学和更高阶的 体外和体内稳态 MΦ 的染色质相互作用。我们的中心假设是 GC 和 IL4 在稳态 MΦ 中创建部分重叠但不同的染色质环境，并且通过充当 GRIP1 是 GR 和 KLF4 的共享辅因子，可促进 M2- 的生理相关功能收敛 就像体内的转录程序一样。我们的具体目标是： 1) 剖析 GRIP1 对 GR 的全球贡献 和 KLF4 增强子形成以及通过全基因组方法形成核心 M2 样转录程序 在离体 GC 和 IL4 极化的 MΦ 中； 2) 评估GRIP1丢失对表型和代谢的影响 体外稳态 MΦ 的特性及其进行表观基因组和转录的能力 编程并支持体内组织修复； 3) 绘制第一张高阶染色质图和 GRIP1- 稳态 MΦ 中依赖的增强子-启动子相互作用，并确定其机制决定因素 GRIP1:KLF4 串扰。该项目的成功完成将产生全面的分析 全局染色质相互作用、核心转录组以及 GRIP1 依赖的分子机制 MΦ 的稳态编程与炎症性 MΦ 激活不同，其定义仍然不明确。