DCAF7/HDAC4/TFEB axis in acute lung injury

DCAF7/HDAC4/TFEB 轴在急性肺损伤中的作用

基本信息

批准号：
10413846
负责人：
Yuan Liu
金额：
$ 47.17万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10413846
关键词：
Acute Acute Lung Injury Acute Respiratory Distress Syndrome Adrenal Cortex Hormones Alveolar Alveolus Attenuated Autophagocytosis Bacteria Bacterial Infections Behavior Bioenergetics Biogenesis Cell Death Cell Nucleus Cells Data Defect Disease Dose Energy Supply Epithelial Epithelial Cells Equilibrium Exhibits FDA approved Functional disorder HDAC4 gene Half-Life Histone Deacetylase Inhibitor Homeostasis Housekeeping Hypoxemia Immune Immunity Impairment Inflammasome Inflammation Inflammatory Inflammatory Response Injury Innate Immune System Libraries Link Lung Lysosomes Mass Spectrum Analysis Mechanical ventilation Mediating Mitochondria Mitochondrial DNA Modality Modeling Molecular Natural Immunity Oxygen Oxygen Consumption Pathway interactions Patients Pattern Peptide Hydrolases Permeability Pharmaceutical Preparations Pneumonia Process Proteins Proteomics Pulmonary Inflammation Reactive Oxygen Species Resolution Role Scheme Severities Signal Transduction Stress Structure of parenchyma of lung Testing Therapeutic Transcription Coactivator Work airway epithelium alveolar epithelium base cytokine cytokine release syndrome improved inhibitor knock-down mitochondrial dysfunction molecular modeling mortality new therapeutic target novel overexpression pathogen preservation prevent protein degradation receptor recruit screening small molecule targeted treatment therapeutic development transcription factor ubiquitin-protein ligase

项目摘要

ABSTRACT Acute Respiratory Distress Syndrome (ARDS) or Acute Lung Injury (ALI) is an acute lung inflammatory process that is associated with an overall mortality ranging from 35% to 50%, and pneumonia is one of the most frequent causes. The central pathophysiology of ARDS/ALI is injury to the epithelium in the airway/alveoli, and overproduction of inflammatory factors in lung tissue. Mitochondria provide the energy supply for numerous cellular activities, and are also the powerhouses of immunity. Bacterial infection impairs mitochondria, and dysfunctional mitochondria need to be cleared through lysosomal degradation, a process termed autophagy/mitophagy. Endogenous damage-associated molecular patterns DAMPs (mtDNA, ATP, ROS) released from damaged mitochondria activate inflammasomes and inflammation. We identify that the deficiency of TFEB, a master transcription factor of autophagy and lysosome biogenesis, impairs mitophagy leading to deleterious DAMP and pro-inflammatory cytokines release, thus activating inflammasomes and inflammation. Through screening a library of FDA-approved drugs (1068 drugs), we identify that the HDAC inhibitor Panobinostat elevates TFEB protein abundance by inhibiting HDAC4. Through unbiased proteomic mass spectrometry analysis, we identify DCAF7 as a substrate receptor for CRL4 ubiquitin E3 ligase recruiting TFEB. DCAF7 overexpression dose-dependently decreases TFEB protein abundance. DCAF7 knockdown prolongs TFEB protein half-life and accumulates its protein levels in the nucleus. Specifically, our preliminary data suggest that 1) bacterial infection decreases TFEB protein levels; 2) TFEB deficiency aggravates harmful DAMP and cytokine release that in turn postpone inflammation resolution; 3) Panobinostat-mediated HDAC4 inhibition preserves TFEB proteins and alleviates bacteria-induced lung inflammation; 4) CRL4 ubiquitin E3 ligase subunit DCAF7 recruits TFEB for proteasomal degradation; 5) a novel small molecule DCAF7 inhibitor BC1753 protects TFEB against degradation. These data led to our hypothesis that inhibiting DCAF7/HDAC4 axis to prevent TFEB from degradation will attenuate bacterial infection-induced lung inflammation and improve inflammation resolution. We will conduct mechanistic studies to determine if TFEB exerts an essential role in bacterial lung inflammation through inhibiting necroptosis (Aim 1). We will examine the role of HDAC4 in regulating TFEB protein stability and lung inflammatory responses in experimental ALI models (Aim 2). We will also test if the small molecule DCAF7 inhibitor alleviates bacterial lung inflammation through preventing TFEB protein degradation (Aim 3). This will be the first study to intervene DCAF7/HDAC4/TFEB axis for inflammatory disease such as ARDS/ALI. Execution of this project will lay the groundwork for a fundamental, paradigm-changing therapeutic advance to regulate innate immunity and treat lung inflammation that will ultimately set the stage for a new translational initiative.

抽象的急性呼吸窘迫综合征（ARDS）或急性肺损伤（ALI）是急性肺与总体死亡率相关的炎症过程范围从35％到50％，并且肺炎是最常见的原因之一。 ARDS/ALI的中央病理生理学是气道/肺泡中的上皮损伤，肺部炎症因子过多生产组织。线粒体为众多细胞活动提供能源供应，也是免疫力。细菌感染会损害线粒体和功能失调线粒体需要通过溶酶体降解来清除，这一过程称为自噬/线粒体。内源性损伤相关的分子模式潮湿（mtDNA，，， ATP，ROS）被损坏的线粒体释放，激活炎症和炎症。我们确定TFEB的缺乏，自噬的主转录因子和溶酶体生物发生，会损害线粒体，导致有害潮湿和促炎性细胞因子释放，从而激活炎症和炎症。通过筛选 FDA批准的药物库（1068种药物），我们确定HDAC抑制剂Panobinostat 通过抑制HDAC4来提高TFEB蛋白丰度。通过公正的蛋白质组学质量光谱分析，我们将DCAF7识别为CRL4泛素E3连接酶的底物受体招募TFEB。 DCAF7过表达剂量依赖性地降低了TFEB蛋白丰度。 DCAF7敲低延长TFEB蛋白半衰期，并在核。具体而言，我们的初步数据表明1）细菌感染降低了TFEB 蛋白质水平； 2）TFEB缺乏症加剧有害潮湿和细胞因子释放推迟炎症分辨率； 3）Panobinostat介导的HDAC4抑制保存TFEB 蛋白质和减轻细菌引起的肺部炎症； 4）CRL4泛素E3连接酶亚基 DCAF7招募蛋白酶体降解的TFEB； 5）一种新型的小分子DCAF7抑制剂 BC1753可保护TFEB免受降解。这些数据导致了我们抑制的假设 DCAF7/HDAC4轴以防止TFEB降解会减弱细菌感染引起的肺部炎症并改善炎症解决方案。我们将进行机械研究确定TFEB是否通过抑制在细菌肺炎症中发挥重要作用坏死作用（AIM 1）。我们将研究HDAC4在调节TFEB蛋白稳定性中的作用实验性ALI模型中的肺炎症反应（AIM 2）。我们还将测试是否小分子DCAF7抑制剂通过预防TFEB来减轻细菌肺炎症蛋白质降解（AIM 3）。这将是第一个干预DCAF7/HDAC4/TFEB轴的研究用于炎症性疾病，例如ARDS/ALI。该项目的执行将为基本的，改变范式的治疗先进，以调节先天免疫和治疗肺部炎症最终将为一项新的翻译计划奠定基础。