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

基本信息

批准号：
10491210
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
项目类别：
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/10491210
关键词：
ATAC-seq Acetyl Coenzyme A Acute Lung Injury Acute Respiratory Distress Syndrome Address 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

项目摘要

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症性血管损伤（例如急性肺损伤（ALI））中恢复取决于肺快速激活组织修复途径（Mφ）的能力，巨噬细胞是最丰富的前哨细胞。肺部的免疫细胞类型是损伤后恢复组织液稳态所必需的，但修复性 Mφ 亚群的身份以及它们是如何产生的仍然难以捉摸。发现了维持肺液稳态所需的肺泡 Mφ (AMφ) 群体的一个子集我们的支持数据表明：1) cAMP 反应丧失。骨髓细胞（CrebΔLyzM 小鼠）中的元素结合 (CREB) 蛋白表达抑制 AMφ 亚群 AMφ 前阶段的谱系，导致 AMφ 生成减少和肺血管通透性增加 2) CrebΔLyzM 小鼠在 LPS 攻击后基本上无法解决损伤，并且在假单胞菌感染后死亡更快铜绿假单胞菌感染；3) 转录组测序 (RNAseq) 和染色质可及性分析 (ATACseq) 流式分选的 CREB-null AMφ 显着改变了基因表达和染色质重塑；4) CREB 是抑制丙酮酸脱氢酶复合物 (PDC) 过度产生核乙酰辅酶 A 所需通过合成丙酮酸脱氢酶激酶 4 (PDK4) 基于这些令人兴奋的初步数据，在目标#1中，我们将检验 CREB 转录促进该屏障分化的假设在 Aim#2 中，我们将研究 CREB 调节修复性 AMφ 的产生。通过控制组蛋白的表观遗传修饰，从而导致 AMφ 基因转录。提出了 CREB 转录活性是上调 PDK4 表达所必需的新概念，这反过来又阻止其靶标 PDC 从线粒体转运至细胞核，从而抑制核乙酰辅酶 A 生成过多并限制组蛋白的表观遗传修饰，导致基因诱导屏障修复 AMφ 群体的转录这些研究将采用严格的多重方法。基因改造小鼠的组学方法（单细胞 RNA、ATAC 和芯片测序）和功能测定包括 Rosa-CrebCxcxr3-ERT 小鼠（其中单核细胞/间质巨噬细胞中有条件删除 CREB）定义 CREB 转录活性在生成修复性 AMφ 群体中的作用。这个屏障修复AMɸ子集是如何产生的应该可以增强这个子集的分化通过药理学或遗传手段造成损伤期间的子集，从而降低 ALI 及相关疾病的死亡率状况。