Quercetin prevents progression of COPD lung disease by modulating Foxo3Aactivity

槲皮素通过调节 Foxo3A 活性预防 COPD 肺部疾病的进展

基本信息

批准号：
9344531
负责人：
Umadevi Sivanappa Sajjan
金额：
$ 42.07万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9344531
关键词：
1-Phosphatidylinositol 3-Kinase Acetylation Affect Animals Anti-Inflammatory Agents Anti-inflammatory Antioxidants Apple Attenuated Autophagocytosis Autophagosome Berry Binding Biological Process Cause of Death Cell Culture Techniques Cell Nucleus Cells Characteristics Chronic Obstructive Airway Disease DNA Binding Data Deacetylase Diet Disease Disease Outcome Disease model Dose E-Cadherin EGF gene EP300 gene Elastases Epidermal Growth Factor Receptor Epithelial Cells FOXO3A gene Flavanol Flavonoids Food Genes Genetic Transcription Goals Goblet Cells Human IL8 gene In Vitro Infection Inflammation Inflammatory Ligands Lung Lung Inflammation Lung diseases Matrix Metalloproteinases Medicine Metaplasia Modeling Molecular Monitor Mucins Mucous body substance Mus Nuclear Nuclear Translocation Onions Outcome Oxidative Stress Patients Phase Phenotype Phosphorylation Phosphotransferases Pilot Projects Plants Predisposition Process Property Protein Phosphatase 2A Regulatory Subunit PR53 Publishing Pulmonary Emphysema Quercetin Resistance Respiratory physiology Rhinovirus SIRT1 gene Surface Testing Transactivation Viral Viral Load result Virus Replication airway epithelium airway inflammation antioxidant enzyme base chemokine cytokine genetic inhibitor improved in vitro Model in vivo inhibitor/antagonist insight knock-down mouse model overexpression p300/CBP-Associated Factor preclinical study prevent promoter public health relevance restoration transcription factor

项目摘要

DESCRIPTION (provided by applicant): Project Summary Quercetin, a plant flavonoid is a potent antioxidant and anti-inflammatory agent. We have performed preclinical studies assessing the suitability of quercetin in the treatment of chronic obstructive pulmonary disease (COPD), a third leading cause of death in the U.S. We showed that quercetin reduces lung inflammation, goblet cell metaplasia and progression of emphysema, in part by increasing expression of the deacetylase Sirt1 in elastase/LPS-exposed mice displaying typical features of COPD. Our pilot studies indicate that elastase/LPS-exposed mice show reduced nuclear FOXO3a, a transcription factor that negatively regulates inflammation and provide resistance to oxidative stress. Further, we found that COPD airway epithelial cells which maintain the phenotypic characteristics of the COPD airway epithelium also show reduced nuclear levels of Sirt1 as well as FOXO3a. Infection with human rhinovirus (RV), a common cause of COPD exacerbations further reduced nuclear Sirt1 and FOXO3a levels. Our pilot studies also show that quercetin promotes translocation of FOXO3a to the nucleus in elastase/LPS-treated mice. Further, quercetin treatment of COPD epithelial cell cultures reduced IL-8 expression and mucus metaplasia, while restoring nuclear FOXO3a levels. Finally, quercetin improved viral clearance after RV challenge in both elastase/LPS mice and COPD epithelial cells. The overall goal is to elucidate the mechanisms by which quercetin modulates nuclear FOXO3a and Sirt1 levels, thereby decreasing inflammation, goblet cell metaplasia and augment viral clearance in COPD models. To accomplish this goal, we propose the following specific Aims. 1. Determine the mechanisms by which quercetin modulates FOXO3a activity and inhibits overexpression of IL-8 and other chemokines and reduces lung inflammation in COPD models. Nuclear translocation of FOXO3a is regulated by phosphorylation and acetylation. We test the hypotheses that 1) quercetin treatment blocks Akt kinase activity, thereby increasing FOXO3A translocation to the nucleus; 2) quercetin-induced Sirt1 deacetylates nuclear FOXO3a, preventing phosphorylation by Akt export from the nucleus; and 3) Sirt1 and FOXO3a together block NF-κB binding to the CXCL-1, 2, 5 and 8 promoter, thereby attenuating cytokine expression and lung inflammation. 2. Examine the mechanisms by which quercetin decreases goblet cell metaplasia in COPD airways. Our pilot data indicate aberrant activation of EGFR in COPD cells as well as in COPD mouse model. We will test the hypotheses that 1) quercetin-induced nuclear FOXO3a positively regulates expression of E-cadherin, which sequesters EGFR at the epithelial cell basolateral surface; 2) quercetin directly inhibits EGFR activity; 3) quercetin, by restoring Sirt1 levels, inhibits matrix metalloproteinase (MMP) expression, thereby decreasing the availability of EGF ligands; and 4) reduced EGFR activity decreases expression of mucin genes, preventing goblet cell metaplasia. 3. Determine the mechanisms by which quercetin increases viral clearance in COPD airways. COPD exacerbations are associated with increased oxidative stress We will test the hypotheses that: 1) oxidative stress induces formation and accumulation of autophagosomes, providing a platform for viral replication; 2) RV infection, via its effects on Akt and CBP/p300, decreases nuclear FOXO3a and phase II antioxidant enzyme expression, amplifying derangements present in COPD cells; and 3) quercetin restores antioxidant enzyme expression by modulation of FOXO3a and Sirt1, decreasing autophagy and increasing viral clearance. Completion of these studies will provide important insights into mechanisms by which quercetin decreases airway inflammation, goblet cell metaplasia and viral clearance in chronic obstructive lung disease. These studies are required to determine the suitability of quercetin as an alternative complimentary medicine in the management of COPD lung disease.

描述（由申请人提供）：项目摘要槲皮素，一种植物类黄酮，是一种有效的抗氧化剂和抗炎剂。我们已经进行了临床前研究，评估槲皮素治疗慢性阻塞性肺病 (COPD) 的适用性，慢性阻塞性肺病是第三个主要原因。我们发现槲皮素可减少肺部炎症、杯状细胞化生和肺气肿的进展，部分是通过增加脱乙酰酶的表达来实现的暴露于弹性蛋白酶/LPS 的小鼠中的 Sirt1 显示出 COPD 的典型特征。我们的初步研究表明，暴露于弹性蛋白酶/LPS 的小鼠的核 FOXO3a 减少，FOXO3a 是一种负向调节炎症并提供对氧化应激的抵抗力的转录因子。维持 COPD 气道上皮表型特征的气道上皮细胞也显示出 Sirt1 和 FOXO3a 感染的核水平降低。人鼻病毒 (RV) 是 COPD 恶化的常见原因，我们的初步研究还表明，槲皮素可促进弹性蛋白酶/LPS 治疗的小鼠中 FOXO3a 易位至细胞核。此外，槲皮素治疗 COPD 上皮细胞。培养物减少了 IL-8 表达和粘液化生，同时恢复了核 FOXO3a 水平，最后，槲皮素提高了 RV 攻击后的病毒清除率。总体目标是阐明槲皮素调节核 FOXO3a 和 Sirt1 水平的机制，从而减少慢性阻塞性肺病模型中的炎症、杯状细胞化生并增强病毒清除。提出以下具体目标 1. 确定槲皮素调节 FOXO3a 活性并抑制 IL-8 和其他物质过度表达的机制。在 COPD 模型中，FOXO3a 的核易位受到磷酸化和乙酰化的调节。我们测试了以下假设：1) 槲皮素治疗阻断 Akt 激酶活性，从而增加 FOXO3A 向细胞核的易位；2) 槲皮素诱导的 Sirt1 去乙酰化核。 FOXO3a，防止 Akt 从细胞核输出磷酸化；3) Sirt1 和 FOXO3a共同阻断 NF-κB 与 CXCL-1、2、5 和 8 启动子的结合，从而减轻细胞因子表达和肺部炎症 2. 检查槲皮素减少 COPD 气道中杯状细胞化生的机制。 COPD 细胞以及 COPD 小鼠模型中的 EGFR 我们将测试以下假设：1) 槲皮素诱导的核 FOXO3a 正向调节 EGFR 的表达。 E-钙粘蛋白，将 EGFR 隔离在上皮细胞基底外侧表面；2) 槲皮素直接抑制 EGFR 活性；3) 槲皮素通过恢复 Sirt1 水平，抑制基质金属蛋白酶 (MMP) 表达，从而降低 EGF 配体的可用性； EGFR 活性降低会降低粘蛋白基因的表达，从而防止杯状细胞化生 3. 确定其机制。槲皮素增加 COPD 气道中的病毒清除率与氧化应激增加有关。我们将测试以下假设：1) 氧化应激诱导自噬体的形成和积累，为病毒复制提供平台；2) RV 感染，通过其对 RV 的影响； Akt 和 CBP/p300，降低核 FOXO3a 和 II 相抗氧化酶表达，放大 COPD 细胞中存在的紊乱；3) 槲皮素通过调节 FOXO3a 和 Sirt1 恢复抗氧化酶的表达，减少自噬和增加病毒清除。这些研究的完成将为了解槲皮素减少慢性阻塞性肺病中气道炎症、杯状细胞化生和病毒清除的机制提供重要见解。需要确定槲皮素作为治疗慢性阻塞性肺疾病的替代补充药物的适用性。