Epigenetic Regulation in Acute Lung Injury

急性肺损伤的表观遗传调控

基本信息

批准号：
8958016
负责人：
Chunbin Zou
金额：
$ 38.5万
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：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8958016
关键词：
Acetylation Acetyltransferase Acute Acute Lung Injury Adult Respiratory Distress Syndrome Affect Affinity Alveolar Alveolar Cell Anticoagulants Apoptosis Argatroban Attenuated Bacteria Bacterial Infections Binding Biology Cartoons Cell Cycle Cell Death Cell Death Induction Cell Survival Cells Cessation of life ChIP-seq Chromatin Complex Computer Simulation Coronary Arteriosclerosis DNA Repair Data Deacetylation Development Docking Dose Enzymes Epigenetic Process Epithelial Epithelial Cells Excision FDA approved Functional disorder Genes Genetic Transcription HDAC1 gene Histone H4 Human Infection Inflammation Inflammatory Label Leucine Zippers Life Link Lipopolysaccharides Lung Lysine Marketing Mediating Messenger RNA Modeling Modification Molecular Morbidity - disease rate Mouse Cell Line Multi-Drug Resistance Mus Necrosis Nosocomial pneumonia Nuclear Organ Orphan Pathogenesis Patients Pharmaceutical Preparations Platelet Factor 4 Play Pneumonia Population Preclinical Drug Evaluation Protein Dephosphorylation Proteins Proteolysis Pseudomonas Infections Pseudomonas aeruginosa Public Health Recruitment Activity Regulation Role Sepsis Severities Signal Pathway Signal Transduction Signal Transduction Pathway Site Structure of parenchyma of lung System Testing Thrombin Ubiquitin Ubiquitination Virulent Work base cell injury cytotoxic cytotoxicity epigenetic regulation histone acetyltransferase improved in vivo inflammatory lung disease inhibitor/antagonist insight malignant breast neoplasm microbial models and simulation mortality multicatalytic endopeptidase complex novel overexpression pathogen pneumocyte protein protein interaction public health relevance respiratory response senescence small hairpin RNA small molecule therapeutic target translational approach ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Acute lung injury and acute respiratory distress syndrome (ALI/ARDS) are severe respiratory illnesses occurring after pneumonia or sepsis. Cell death is a hallmark of ALI/ARDS. Pseudomonas aeruginosa is a major bacterial isolate from pneumonia that rapidly develops multi-drug resistance in subjects who develop ALI/ARDS. The underlying mechanism (s) of how epigenetics affects the pathobiology of ARDS is unknown. Our exciting preliminary results demonstrate that a multifunctional chromatin modulator, Mortality factor 4 like 1 (Morf4l1) epigenetically induces cell death in response to P. aeruginosa infection. Morf4l1 protein levels are elevated in human pneumonia lung tissue and in lipopolysaccharide (LPS) treated or P. aeruginosa infected lung epithelial cells. Interestingly, Morf4l1 is a highly cytotoxic protein that is rapidly eliminated in cells by an orphan ubiquitin E3 ligase subunit calld Fbxl18. However, P. aeruginosa infection protects Morf4l1 from degradation by increasing Morf4l1 acetylation that antagonizes its ubiquitination and removal in cells. Overexpression of Morf4l1 leads to profound epithelial cell death and is lethal to mice. Preliminary data also suggest that P. aeruginosa infection enhances Morf4l1 nuclear entry that may allow it escape its cytosolic degradation to facilitate its epigenetic function to control inflammatory gene transcription. Depletion of Morf4l1 by shRNA in mice effectively represses P. aeruginosa induced cell death. Importantly, by in silico drug screening, we found an effective Morf4l1 antagonist, argatroban, an FDA approved anticoagulant currently in the market. Argatroban tightly binds to Morf4l1, inhibits its epigenetic function, reduces Morf4l1 mediated cell death, an promotes cell survival during P. aeruginosa infection. In addition, administration of argatroban improves survival, suppresses cell death, and ameliorates inflammation in experimental murine models of ALI. These data led to our hypothesis that Pseudomonas infection protects Morf4l1 from SCF-Fbxl18 mediated degradation through its subcellular relocation and modification, thereby epigenetically modulating inflammation and triggering cell death. As a corollary to this hypothesis, we will test if targeting Morf4l1 with an FDA approved drug may attenuate the severity of experimental ALI. We will test this hypothesis by executing two specific aims: (1): to determine if P. aeruginosa infection triggers signal transduction to activate lysine acetyltransferase Gcn5 to interact and catalyze Morf4l1 acetylation, and the acetylation increases Morf4l1 protein levels that protects it from E3 ubiquitin ligase SCFFbxl18 mediated ubiquitin-proteasomal proteolysis. (2): to identify if P. aeruginosa infection triggers Morf4l1 dephosphorylation that enhances Morf4l1 nuclear entry to leads to epigenetic changes and cell death. Last we will target Morf4l1 with a small molecule (argatroban) to attenuate acute lung injury. These studies will provide a significant conceptual advance regarding epigenetic regulation in the pathogenesis of ALI/ARDS. The project also provides new insight into the molecular control of a poorly described chromatin modulator (Morf4l1) in the regulation of cell death and its pathological role in ALI/ARDS. The study identifies an off-labeling effect of an FDA approved drug that paves a new way for pharmacotherapeutic targeting of Morf4l1 in ALI/ARDS pathogenesis.

描述（由申请人提供）：急性肺损伤和急性呼吸窘迫综合征 (ALI/ARDS) 是肺炎或败血症后发生的严重呼吸系统疾病，是 ALI/ARDS 的标志，是从肺炎中分离出来的主要细菌。发生 ALI/ARDS 的受试者会迅速产生多重耐药性表观遗传学如何影响 ARDS 病理学的潜在机制尚不清楚。死亡因子 4 样 1 (Morf4l1) 是一种多功能染色质调节剂，可表观遗传地诱导细胞死亡，以响应铜绿假单胞菌感染。 Morf4l1 蛋白水平在人肺炎肺组织和脂多糖 (LPS) 或铜绿假单胞菌感染的肺上皮细胞中升高，Morf4l1 是一种高细胞毒性蛋白，可被称为 Fbxl18 的孤儿泛素 E3 连接酶亚基快速消除。 , 铜绿假单胞菌感染通过增加 Morf4l1 乙酰化来保护 Morf4l1 免遭降解拮抗 Morf4l1 在细胞中的泛素化和去除，导致严重的上皮细胞死亡，并且对小鼠是致命的。初步数据还表明，铜绿假单胞菌感染增强了 Morf4l1 的核进入，可能使其逃避胞质降解，从而促进其表观遗传功能。控制小鼠炎症基因转录。通过 shRNA 消除 Morf4l1 可有效抑制铜绿假单胞菌诱导的细胞死亡。重要的是，通过计算机药物筛选，我们发现了一种有效的 Morf4l1 拮抗剂阿加曲班，这是一种 FDA 批准的目前市场上的抗凝剂，阿加曲班与 Morf4l1 紧密结合，抑制其表观遗传功能，减少 Morf4l1 介导的细胞死亡，并促进疟原虫感染期间的细胞存活。此外，给予阿加曲班可提高实验小鼠的存活率、抑制细胞死亡并改善炎症。这些数据得出了我们的假设，即假单胞菌感染通过其亚细胞重定位和修饰保护 Morf4l1 免受 SCF-Fbxl18 介导的降解，从而表观遗传调节炎症并触发细胞死亡。作为该假设的推论，我们将测试是否针对 Morf4l1。使用 FDA 批准的药物可能会减轻实验性 ALI 的严重程度，我们将通过执行两个具体目标来检验这一假设：(1)：确定 P.铜绿假单胞菌感染触发信号转导，激活赖氨酸乙酰转移酶 Gcn5 相互作用并催化 Morf4l1 乙酰化，乙酰化增加 Morf4l1 蛋白水平，保护其免受 E3 泛素连接酶 SCFFbxl18 介导的泛素蛋白酶体蛋白水解。 Morf4l1 去磷酸化可增强 Morf4l1 进入核，从而导致表观遗传变化和细胞死亡。最后，我们将用小分子（阿加曲班）靶向 Morf4l1，以减轻急性肺损伤。这些研究将为 ALI 发病机制中的表观遗传调控提供重要的概念进展。 /ARDS 还为一种鲜为人知的染色质调节剂 (Morf4l1) 在细胞死亡调节中的分子控制提供了新的见解。该研究确定了 FDA 批准的药物的标签外效应，为针对 ALI/ARDS 发病机制的 Morf4l1 药物治疗开辟了新途径。