Role of FABP5 in COPD Exacerbations

FABP5 在 COPD 恶化中的作用

• 批准号: 10397397
• 负责人: Fabienne Gally
• 金额: $ 49.54万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10397397
• 关键词: Acute; Affect; Alveolar Macrophages; Anti-Inflammatory Agents; Apoptotic; Bacterial Infections; Binding; Biological Assay; Bronchitis; Cause of Death; Cells; Chronic Obstructive Pulmonary Disease; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Diagnosis; Disease; Early Intervention; Economic Burden; Enhancers; Excision; Fatty Acids; Feedback; Flare; Frequencies; Future; Gene Expression; Genes; Genomic approach; Goals; Hemophilus influenza infection; Homeostasis; Impairment; In Vitro; Individual; Infection; Inflammation; Inflammatory; Intervention; JUN gene; Link; Lung; Mediating; Messenger RNA; Modeling; Molecular; Mononuclear; Morbidity - disease rate; Mus; Mutagenesis; Nature; Nontypable Haemophilus influenza; Nuclear Receptors; PPAR gamma; Pathogenesis; Patients; Peripheral Blood Mononuclear Cell; Phagocytes; Pneumonia; Prevalence; Prevention strategy; Process; Proteins; Pulmonary Inflammation; Recording of previous events; Recurrence; Reporter; Reporting; Research; Resolution; Respiratory Tract Infections; Risk Factors; Role; Sampling; Site; Smoker; Structure of parenchyma of lung; Suppressor Genes; Testing; Therapeutic; United States; Viral; Wild Type Mouse; Work; base; cigarette smoke; cigarette smoking; clinically relevant; clinically significant; exposure to cigarette smoke; fatty acid-binding proteins; genetic approach; improved; in vivo; macrophage; monocyte; mortality; mouse model; novel; novel strategies; overexpression; oxidation; pre-clinical; prevent; programs; promoter; protein expression; public health relevance; recruit; repaired; response

PROJECT SUMMARY Chronic Obstructive Pulmonary Disease (COPD) is the third leading cause of death in the United States and its global prevalence continues to rise due to lack of an effective cure. Although cigarette smoke (CS) is by far the most important risk factor of COPD, repeated and sustained infections are clearly linked to disease pathogenesis (e.g., exacerbations). Interestingly, COPD exacerbations are a risk factor for having more exacerbations, suggesting that a vicious cycle of ineffective resolution of inflammation predisposes individuals for future episodes. Thus, identifying patients likely to have recurrent exacerbations may allow early implementation of appropriate preventive strategies and would be an important new approach in the treatment of the disease. We have discovered that Fatty Acid Binding Protein 5 (FABP5) is important for the resolution of inflammation. However we do not fully comprehend the mechanisms behind FABP5 protective role in COPD exacerbations. In this proposal we will focus on FABP5 in mononuclear phagocytes because these cells are at the center of the processes required for the resolution of inflammation, including engulfment of apoptotic cells and activation of anti-inflammatory nuclear receptors such as Peroxisome Proliferator-Activated Receptor γ (PPARγ). Our preliminary studies indicate impaired apoptotic cell engulfment by alveolar macrophages and reduced monocyte recruitment following bacterial infection in FABP5-/- mice. In addition, FABP5-/- mice have increased bacterial and viral-induced lung inflammation, compared to wild type (WT) mice. The clinical significance of these findings is supported by decreased FABP5 mRNA and protein levels in peripheral blood mononuclear cells (PBMCs) of COPD patients, especially among those with a history of episodes of bronchitis, pneumonia or COPD exacerbations and by the discovery of new SNPs in the FABP5 gene that are associated with severe exacerbations. We hypothesize that decreased FABP5 following CS exposure worsens infection- induced lung inflammation by preventing mononuclear lung phagocyte reprogramming, which is necessary for resolution of lung inflammation in COPD exacerbations. To test this hypothesis, we propose 3 specific aims. Under specific aim 1, we will test the hypothesis that CS impairs FABP5-mediated mononuclear lung phagocytes recruitment and efferocytosis. Using WT, FABP5-/- or FABP5-/- mice complemented with FABP5 in mononuclear phagocytes we will determine how FABP5 expression affects mononuclear phagocyte recruitment in response to infection in the presence or absence of CS. We will also determine how FABP5 expression affects mononuclear phagocyte engulfment capabilities following CS and/or infection in vivo and in vitro. Under aim 2, we will test the hypothesis that FABP5/PPARγ interaction is necessary to promote resolution of inflammation and return to homeostasis and determine whether CS impairs this interaction. We will manipulate FABP5 levels to demonstrate that the presence of FABP5 in mononuclear phagocytes is essential for PPARγ-mediated activation and functions. These functions include fatty acid β-oxidation and macrophage alternative programming for processing of engulfed materials. We will also determine whether PPARγ increases FABP5 expression as a positive feedback loop using ChIP, silencing, promoter reporter assay and mutagenesis. Under specific aim 3, we will test the hypothesis that the newly identified FABP5 SNPs associated with COPD exacerbations and alveolar macrophages exposed to CS have less FABP5 expression, less engulfment capabilities and less PPARγ activity. We propose to analyze FABP5 gene and protein expression using our existing samples from patients with or without the newly identified SNPs or alveolar macrophages from smokers and see whether FABP5 expression level correlates with SNP expression, CS exposure, and COPD exacerbations. Additionally, we propose to characterize the different regions encompassing the SNPs for their enhancer or suppressor gene activity and to dissect their enhancer function using multiplex CRISPR-based enhancer interference. And finally, we propose to look at inflammatory versus pro-resolving markers and efferocytosis capabilities of those cells. The completion of these aims will provide an in-depth understanding of the mechanisms by which CS inhibits FABP5 function and leads to over inflamed lung tissue. Ultimately, the goal of this research is to improve the preclinical diagnosis and treatment of COPD exacerbations, and thus could have a highly significant overall impact on what is currently an incurable disease.

项目总结 慢性阻塞性肺疾病(COPD)是美国第三大死因 由于缺乏有效的治疗方法，其全球流行率继续上升。虽然香烟烟雾(CS)是由 作为慢性阻塞性肺疾病最重要的危险因素，反复和持续的感染显然与疾病有关 发病机制(例如，病情恶化)。有趣的是，慢性阻塞性肺病的恶化是一个危险因素 病情恶化，表明炎症消解无效的恶性循环使个人 为未来的剧集做准备。因此，识别可能有复发恶化的患者可能有助于及早 实施适当的预防战略，并将是治疗的重要新方法 这种疾病的危害。我们发现，脂肪酸结合蛋白5(FABP5)对蛋白质的分解非常重要。 发炎。然而，我们并不完全理解FABP5在COPD中保护作用的机制 病情加重。在这项提案中，我们将重点研究单核吞噬细胞中的FABP5，因为这些细胞处于 炎症消退所需过程的中心，包括吞噬凋亡细胞 和抗炎核受体的激活，如过氧化体增殖物激活受体γ (PPARγ)。 我们的初步研究表明，肺泡巨噬细胞和 FABP5-/-小鼠在细菌感染后单核细胞募集减少。此外，FABP5-/-小鼠有 与野生型(WT)小鼠相比，细菌和病毒诱导的肺部炎症增加。临床部 外周血液中FABP5的mRNA和蛋白水平降低支持了这些发现的意义 COPD患者的单个核细胞(PBMC)，特别是有支气管炎病史的患者， 肺炎或COPD恶化以及FABP5基因中新的SNPs的发现与此相关 病情严重恶化。我们假设在CS暴露后FABP5的减少会加重感染- 通过防止单核肺吞噬细胞重编程来诱导肺部炎症，这是 慢性阻塞性肺疾病急性加重期肺部炎症的消退。为了验证这一假设，我们提出了三个具体目标。 在特定目标1下，我们将检验CS损害FABP5介导的单核肺吞噬细胞的假设 募集和泡沫化。用WT、FABP5-/-或FABP5-/-小鼠与FABP5互补的单核小鼠 吞噬细胞我们将确定FABP5的表达如何影响单核巨噬细胞的募集反应 在CS存在或不存在的情况下对感染的敏感性。我们还将确定FABP5的表达如何影响 体内和体外CS和/或感染后单核吞噬细胞的吞噬能力。在目标2下， 我们将验证FABP5/PPARγ相互作用对于促进炎症消退是必要的假设 并回到稳态，并确定CS是否损害了这种相互作用。我们将操纵FABP5级别 为了证明单核吞噬细胞中FABP5的存在是PPARγ介导的必需的 激活和功能。这些功能包括脂肪酸β氧化和巨噬细胞替代编程 用于处理被吞没的材料。我们还将确定PPARγ是否会增加FABP5的表达 正反馈环使用芯片、沉默、启动子报告实验和突变。在具体目标3下， 我们将检验这样一种假设，即新发现的FABP5 SNPs与COPD恶化和 暴露于CS的肺泡巨噬细胞FABP5表达减少，吞噬能力减弱，PPARγ减少 活动。我们建议使用我们现有的患者样本来分析FABP5基因和蛋白的表达 不管有没有新发现的SNPs或吸烟者的肺泡巨噬细胞，看看FABP5 表达水平与SNP表达、CS暴露和COPD恶化相关。此外，我们 建议表征围绕SNPs的增强基因或抑制基因的不同区域 并使用基于CRISPR的多重增强子干扰来剖析其增强子功能。最后， 我们建议观察这些细胞的炎性标志物和促分解标志物以及吞噬能力。 这些目标的完成将使我们深入了解CS通过哪些机制 抑制FABP5功能，导致肺组织过度炎症。归根结底，本研究的目标是改进 COPD病情加重的临床前诊断和治疗，因此可能在总体上具有非常重要的意义 对目前不治之症的影响。

项目成果

Altered Macrophage Function Associated with Crystalline Lung Inflammation in Acid Sphingomyelinase Deficiency.

酸性鞘磷脂酶缺乏症中与结晶性肺炎症相关的巨噬细胞功能改变。

• DOI: 10.1165/rcmb.2020-0229oc
• 发表时间: 2021
• 期刊: American journal of respiratory cell and molecular biology
• 影响因子: 6.4
• 作者: Poczobutt,JoannaM;Mikosz,AndrewM;Poirier,Christophe;Beatman,EricaL;Serban,KarinaA;Gally,Fabienne;Cao,Danting;McCubbrey,AlexandraL;Cornell,ChristinaF;Schweitzer,KellyS;Berdyshev,EvgenyV;Bronova,IrinaA;Paris,François;Petra
• 通讯作者: Petra

Macrophage programming is regulated by a cooperative interaction between fatty acid binding protein 5 and peroxisome proliferator-activated receptor γ.

• DOI: 10.1096/fj.202200128r
• 发表时间: 2022-05
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology