CREB Instruction of Macrophage Fate and Lung fluid homeostasis

CREB对巨噬细胞命运和肺液稳态的指导

• 批准号: 10305990
• 负责人: DOLLY MEHTA
• 金额: $ 15.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10305990
• 关键词: ATAC-seq; Acetyl Coenzyme A; Acute Lung Injury; Address; Adult Respiratory Distress Syndrome; Alveolar; Alveolar Macrophages; BACH2 gene; Binding; Biological Assay; Cell Nucleus; Cells; ChIP-seq; Chromatin; Critical Illness; Cyclic AMP; Cyclic AMP Response Element; Cyclic AMP-Responsive DNA-Binding Protein; Data; Development; Disease; Endothelium; Epigenetic Process; Epithelial; Exhibits; Extravasation; Fluid Balance; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Histone Acetylation; Homeostasis; ITGAM gene; ITGAX gene; Immune; Impairment; Inflammatory; Inflammatory Response; Injury; Instruction; Leukocytes; Life; Lung; Lung Capacity; Mitochondria; Mus; Myeloid Cells; Nuclear; Pathway interactions; Patients; Pharmacology; Plasma; Population; Production; Pseudomonas aeruginosa infection; Pyruvate Dehydrogenase Complex; RNA; Recovery; Regulation; Regulatory Element; Research; Role; Sentinel; Signal Pathway; Signal Transduction; Source; Testing; Therapeutic; Tissues; Transposase; Vascular Permeabilities; XCL1 gene; base; cell type; chromatin remodeling; cytokine; histone modification; injury and repair; insight; interstitial; lung injury; lung repair; lung vascular injury; macrophage; monocyte; mortality; multiple omics; novel; novel therapeutic intervention; prevent; programs; promoter; protein expression; pyruvate dehydrogenase kinase 4; repair strategy; restoration; single-cell RNA sequencing; tissue injury; tissue repair; transcription factor; transcriptome sequencing; vascular injury

Abstract Recovery from severe forms of inflammatory vascular injury, such as acute lung injury (ALI), depends on the lung's capacity to rapidly activate tissue repair pathways. Macrophages (Mφ), the most abundant sentinel immune cell type in the lung, are required for restoration of tissue-fluid homeostasis following injury, but the identity of the reparative Mφ subpopulations and how they are generated remain elusive. Here, we have discovered a subset of the alveolar Mφ (AMφ) population that is required to maintain lung fluid homeostasis basally and induce tissue repair after injury. Our Supporting Data show that: 1) loss of cAMP Response Element Binding (CREB) protein expression in myeloid cells (Creb∆LyzM mice) arrest a subpopulation of the AMφ lineage at the pre-AMφ stage, leading to decreased AMφ generation and increased lung vascular permeability basally; 2) Creb∆LyzM mice failed to resolve injury post-LPS challenge and died more rapidly after Pseudomonas aeruginosa infection; 3) transcriptome sequencing (RNAseq) and chromatin accessibility profiling (ATACseq) of flow-sorted CREB-null AMφ have markedly altered gene expression and chromatin remodeling; and 4) CREB is required to inhibit excessive production of nuclear acetyl-CoA from the pyruvate dehydrogenase complex (PDC) through synthesis of pyruvate dehydrogenase kinase 4 (PDK4). Based on these provocative Preliminary Data, in Aim#1, we will test the hypothesis that CREB transcriptionally promotes differentiation of this barrier reparative AMφ subpopulation. In Aim#2, we will investigate that CREB regulates reparative AMφ generation by controlling epigenetic modifications of histones thereby leading to AMφ gene transcription. Here, we will address the novel concept that the transcriptional activity of CREB is required to upregulate PDK4 expression, which in turn prevents transport of its target, PDC, from mitochondria to the nucleus thereby suppressing excessive nuclear acetyl Co-A generation and limiting epigenetic modifications of histones, leading to gene transcription that induces the barrier reparative AMφ population. These studies will employ a rigorous multi- omics approach (single-cell RNA-, ATAC-and Chip-sequencing) and functional assays in genetically altered mice including Rosa-CrebCxcxr3-ERT mice (in which CREB is conditionally deleted in monocytes/interstitial macrophages) to define the role of CREB transcriptional activity in generating the reparative AMφ population. Understanding how this barrier reparative AMɸ subset is generated should make it possible to enhance differentiation of this subset during injury by pharmacological or genetic means, thereby reducing the mortality of ALI and related conditions.

抽象的 从严重形式的炎性血管损伤（例如急性肺损伤（ALI））中恢复，取决于 肺部快速激活组织修复途径的能力。巨噬细胞（Mφ），最丰富的哨兵 肺中的免疫细胞类型是恢复受伤后组织流体稳态需要的，但是 修复Mφ亚群的身份及其如何产生仍然难以捉摸。在这里，我们有 发现了维持肺稳态所需的肺泡Mφ（AM或）种群的子集 基本并在受伤后诱导组织修复。我们的支持数据表明：1）营地反应丧失 髓样细胞（CREBΔlyzm小鼠）中的元素结合（CREB）蛋白表达阻碍了Amφ的亚群 谱系在前AMφ阶段，导致AMφ产生降低并增加了肺血管通透性 基本2）CrebΔlyzm小鼠无法解决LPS挑战后损伤，并在假单胞菌后迅速死亡 铜绿节感染； 3）转录组测序（RNASEQ）和染色质可及性分析（ATACSEQ） 流动的CREB-NULLAMφ显着改变了基因表达和染色质重塑。 4）Creb是 需要抑制丙酮酸脱氢酶复合物（PDC）的过量产生核乙酰-COA 通过丙酮酸脱氢酶激酶4（PDK4）的合成。基于这些挑衅性的初步数据， 在AIM＃1中，我们将测试CREB转录促进该障碍的分化的假设 修复性AMφ亚群。在AIM＃2中，我们将调查CREB调节修复Amφ的生成 通过控制组蛋白的表观遗传修饰，从而导致AMφ基因转录。在这里，我们会的 解决新的概念，即需要CREB的转录活动才能更新PDK4表达式， 反过 过度的核乙酰基共乙酰基共同生成和限制组蛋白的表观遗传修饰，导致基因 诱导屏障修复型AM或种群的转录。这些研究将采用严格的多种多样 OMICS方法（单细胞RNA-，ATAC和芯片序列）和一般改变的小鼠的功能测定 包括Rosa-Crebcxcxr3-ert小鼠（其中CREB在单核细胞/间质巨噬细胞中有条件地删除） 定义CREB转录活性在产生修复amφ种群中的作用。理解 如何产生这种障碍频率的AMɸ子集应使得增强这种差异化 药物或遗传手段受伤期间的子集，从而降低了ALI的死亡率和相关的死亡率 状况。