HDAC6 regulates cigarette smoke-induced endothelial barrier dysfunction and lung injury

HDAC6 调节香烟烟雾引起的内皮屏障功能障碍和肺损伤

• 批准号: 9285844
• 负责人: Qing Lu
• 金额: $ 32.18万
• 依托单位: OCEAN STATE RESEARCH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9285844
• 关键词: Acute Lung Injury; Address; Adult Respiratory Distress Syndrome; Adverse effects; Affect; Animal Model; Bacterial Infections; Binding; Blood Circulation; Blood Vessels; Cell Culture Techniques; Cell physiology; Cessation of life; Characteristics; Data; Deacetylase; Deacetylation; Disease; Dynamin; Endothelial Cells; Endothelium; Epidemiology; Functional disorder; Goals; HDAC6 gene; Human; Impairment; In Vitro; Individual; Infection; Injury; Knockout Mice; Life; Link; Lung; Maintenance; Mediating; Mediator of activation protein; MicroRNAs; Microtubule-Associated Proteins; Microtubules; Mission; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Morbidity - disease rate; Organ; Oxidants; Pattern; Permeability; Phosphorylation; Population; Predisposition; Preventive; Proteins; Pseudomonas; Pseudomonas Infections; Pseudomonas aeruginosa; Public Health; Quality Control; Reactive Oxygen Species; Research; Research Personnel; Respiratory Tract Infections; Risk Factors; Role; Smoker; Syndrome; Technology; Therapeutic; Tubulin; United States National Institutes of Health; alpha Tubulin; cigarette smoke-induced; cigarette smoking; clinical risk; environmental tobacco smoke exposure; epidemiology study; in vivo; innovation; insight; lung injury; mitochondrial dysfunction; mortality; novel; novel therapeutic intervention; novel therapeutics; oxidation; pre-clinical; premature; prevent; segregation

Cigarette smoke (CS) affects 1 billion people worldwide and causes 6 million premature deaths from associated diseases each year. Acute lung injury (ALI)/ acute respiratory distress syndrome (ARDS) is a life- threatening condition with 40% mortality. Emerging epidemiology studies have shown that CS predisposes lungs to infections and increases the likelihood of ALI/ ARDS. However, the mechanism of adverse effects of CS on ALI is unknown. There is an urgent need for strategies to prevent and treat ALI/ ARDS in smokers. The objectives of this application is to use human lung microvascular endothelial cells (LMVEC) and preclinical animal models to identify mechanisms by which CS impairs pulmonary endothelial barrier function and increases vulnerability to ALI after bacterial infection and to develop strategies to prevent and restore endothelial barrier function and ALI in susceptible populations. This study uses state-of-art technologies and comprehensive approaches to address a central hypothesis that CS increases endothelial cell permeability and predisposes lungs to ALI after infection by a mechanism in which oxidants from CS and mitochondria cause Akt oxidation and subsequent GSK3β-dependent activation of histone deacetylase 6 (HDAC6), this results in microtubule deacetylation, leading to translocation of dynamin-related protein (Drp)1 to mitochondria, culminating in mitochondrial fission and dysfunction and subsequent release of mitochondrial damage- associated molecular patterns (mtDAMPs). Aim 1: We will first determine the role of HDAC6 in CS priming for ALI after Pseudomonas aeruginosa infection by using global HDAC6 knockout mice, endothelium-specific lentiviral HDAC6 miRNA silencing in vivo, and LMVEC isolated from HDAC6 knockout mice; we will then determine the mechanism by which CS exposure leads to HDAC6 activation. Aim 2: We will determine the role of HDAC6 in CS-induced mitochondrial fission and dysfunction and their roles in mediating increased endothelial permeability and priming for ALI after P. aeruginosa infection. We will first characterize the effects of CS exposure on Drp1 translocation and on mitochondrial fission, fusion, function, and release of mtDNA in vitro and in vivo; we will then determine the effect of blocking HDAC6 or α-tubulin deacetylation on the adverse effects of CS on mitochondria in vitro and in vivo; finally, we will determine the roles of mitochondrial fission, mitochondrial ROS and mtDAMPs in CS-induced increased endothelial permeability in vitro and CS priming for ALI in vivo. This study will provide innovative insights into the effect of CS on lung endothelial barrier function. Establishment of the links among HDAC6, mitochondrial fission and mtDAMPs may result in new therapeutic approaches to CS-induced vascular injury in the lung and potentially also in the systemic circulation.

香烟烟雾 (CS) 影响着全世界 10 亿人，并导致 600 万人过早死亡 每年都会发生相关疾病。急性肺损伤（ALI）/急性呼吸窘迫综合征（ARDS）是一种致命的疾病 死亡率为 40% 的威胁状况。新兴流行病学研究表明，CS 易诱发 肺部感染并增加 ALI/ARDS 的可能性。然而，不良反应的机制 ALI 上的 CS 未知。迫切需要预防和治疗吸烟者 ALI/ARDS 的策略。这 本申请的目标是使用人肺微血管内皮细胞 (LMVEC) 和临床前 动物模型以确定 CS 损害肺内皮屏障功能的机制 细菌感染后增加 ALI 的脆弱性并制定预防和恢复策略 易感人群的内皮屏障功能和 ALI。这项研究使用了最先进的技术和 解决 CS 增加内皮细胞通透性这一中心假设的综合方法 并通过来自 CS 和线粒体的氧化剂的机制使肺部在感染后易患 ALI 导致 Akt 氧化和随后 GSK3β 依赖性的组蛋白脱乙酰酶 6 (HDAC6) 激活，这 导致微管脱乙酰化，导致动力相关蛋白 (Drp)1 易位至线粒体， 最终导致线粒体裂变和功能障碍以及随后释放的线粒体损伤- 相关分子模式（mtDAMP）。目标 1：我们首先确定 HDAC6 在 CS 启动中的作用 使用全局 HDAC6 敲除小鼠，内皮特异性，铜绿假单胞菌感染后发生 ALI 体内慢病毒 HDAC6 miRNA 沉默，以及从 HDAC6 敲除小鼠中分离出的 LMVEC；然后我们会 确定 CS 暴露导致 HDAC6 激活的机制。目标 2：我们将确定 HDAC6 在 CS 诱导的线粒体分裂和功能障碍中的作用及其在介导增加中的作用 铜绿假单胞菌感染后内皮通透性和 ALI 启动。我们首先将描述效果 CS 暴露对 Drp1 易位以及线粒体裂变、融合、功能和 mtDNA 释放的影响 体外和体内；然后我们将确定阻断 HDAC6 或 α-微管蛋白脱乙酰化对不良反应的影响 CS对体外和体内线粒体的影响；最后，我们将确定线粒体裂变的作用， CS 诱导的体外内皮通透性增加和 CS 启动中的线粒体 ROS 和 mtDAMP 体内ALI。这项研究将为 CS 对肺内皮屏障功能的影响提供创新的见解。 HDAC6、线粒体裂变和 mtDAMP 之间联系的建立可能会带来新的治疗方法 治疗 CS 引起的肺部血管损伤以及可能的全身循环血管损伤的方法。