Dysregulation of the NFkB/Sox18 Axis During Ventilator-Induced Lung Injury (VILI)

呼吸机引起的肺损伤 (VILI) 期间 NFkB/Sox18 轴失调

• 批准号: 10094246
• 负责人: Stephen M Black
• 金额: $ 43.35万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-05 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10094246
• 关键词: Acute Lung Injury; Adult Respiratory Distress Syndrome; Attenuated; Candidate Disease Gene; Capillary Endothelial Cell; Critical Care; Critical Illness; DNA Methylation; Data; Development; Down-Regulation; Endothelial Cells; Endothelium; Epigenetic Process; Event; Exhibits; Exposure to; Extravasation; Functional disorder; Genetic; Goals; Histone Deacetylase; Histone Deacetylase Inhibitor; Human; I Kappa B-Alpha; Inflammation Mediators; Inflammatory; Intercellular Junctions; Intervention; Life; Lipopolysaccharides; Liquid substance; Lung; Lung diseases; Mechanical Stress; Mechanical ventilation; Mediating; Mitochondria; Modification; Mus; Nuclear; Oxidative Stress; Patients; Peptides; Periodicity; Permeability; Phase; Phosphorylation; Post-Translational Protein Processing; Pre-Clinical Model; Predisposition; Proteins; Publishing; Reactive Oxygen Species; Reagent; Refractory; Regulation; Respiratory Failure; Role; SOX18 gene; Savings; Signal Transduction; Signaling Protein; Single Nucleotide Polymorphism; Stretching; Syndrome; Testing; Therapeutic; Tidal Volume; Tight Junctions; Transcriptional Regulation; Translating; Treatment Efficacy; United States; Ventilator-induced lung injury; cytokine; improved; individualized medicine; inflammatory lung disease; insight; lung injury; mechanical force; mortality; new therapeutic target; nicotinamide phosphoribosyltransferase; nitration; novel; preservation; prevent; promoter; receptor; recruit; response; shear stress; synergism; therapeutic target; transcription factor

PROJECT SUMMARY: Mechanical ventilation creates excessive mechanical stress that directly augments lung injury in critically ill patients with respiratory failure due to ARDS, a syndrome known as ventilator-induced lung injury (VILI). There have been little insights into the specific mechanisms involved in VILI-induced pulmonary capillary endothelial cell (EC) leakage, a key therapeutic target in ARDS/VILI. Project #1 will examine how excessive mechanical stress suppresses SOX18 expression, a transcription factor (TF) intimately involved in preservation of EC barrier integrity via transcriptional regulation of the tight junction protein, claudin 5 (CLDN5). We have shown that fluid shear stress increases Sox18 expression, however, EC exposed to 18% cyclic stretch (CS) or LPS exhibit decreased Sox18 expression. In addition, enhanced mouse lung Sox18 expression attenuates EC barrier disruption induced by exposure to high tidal volume mechanical ventilation or LPS. Project #1 will investigate mechanisms by which LPS and excessive mechanical stress reduce Sox18 expression in both human lung EC and the mouse lung. We will explore how the initial inflammatory phase of VILI, via inflammatory signaling evoked by nuclear factor (NF)-κB, suppresses SOX18 expression. Further, Project #1 will extend our published data and examine how excessive mechanical stress regulates NF-κB signaling proteins via increased ROS- mediated post-translational NF-κB modifications (PTMs) including nitration and phosphorylation. These events, in concert with recruitment of histone deacetylases, appear to be critical to suppressing SOX18 promoter activity. Specific Aim #1 (SA #1) will define the role of NF-κB signaling in mechanical stress-mediated downregulation of SOX18 and CLDN5 expression and loss of EC junctional integrity. SA #1 will also investigate the influence of ARDS/VILI related SOX18 and CLDN5 promoter SNPs on Sox18 and CLDN5 expression in response to mechanical stress. SA #2 will define the role of nitration-mediated PTMs in the mechanical stress- mediated activation of NF-κB signaling and investigate the involvement of the mechanosensitive receptor, TRPV4. SA #3 will translate SAs #1 & 2 data into actionable information and define the therapeutic efficacy of directly increasing Sox18 expression, preventing the epigenetic down-regulation of SOX18 (HDAC inhibitors), preventing nitration-mediated NF-κB activation (shielding peptides) and reducing mitochondrial-derived ROS (mitochondrial targeted peptides). Together with each PPG Project and the expertise residing in each PPG Core, these explorations will advance our understanding of the interplay between genetics, epigenetics and PTMs in regulating the SOX18-CLDN5 axis and EC barrier integrity. Further, our studies will promote a more thorough understanding of VILI pathobiology while highlighting the application of individualized therapies for the critically ill.

项目总结： 机械通风产生过多的机械应力，直接加重危重病患者的肺损伤 由于ARDS导致呼吸衰竭的患者，这种综合征被称为呼吸机诱导肺损伤(VILI)。那里 对VILI诱导的肺毛细血管内皮细胞参与的具体机制知之甚少 细胞(EC)渗漏是ARDS/VILI的关键治疗靶点。项目1将检查过度机械 应激抑制与EC保存密切相关的转录因子SOX18的表达 通过紧密连接蛋白CLDN5(CLDN5)的转录调节，屏障的完整性。我们已经证明了 流体剪切力增加Sox18的表达，然而，暴露于18%循环拉伸(CS)或内毒素的EC表现出 降低Sox18的表达。此外，增强的小鼠肺组织Sox18表达可减弱EC屏障 暴露于大潮气量机械通气机或脂多糖引起的干扰。项目1将调查 内毒素和过度机械应激降低人肺内皮细胞SOX18表达的机制 还有老鼠的肺。我们将探索VILI的初始炎症阶段是如何通过炎症信号 受核因子-κB诱导，抑制SOX18的表达。此外，项目1将延长我们发布的 数据，并研究过度机械应力如何通过增加ROS来调节NF-κB信号蛋白。 介导的翻译后核因子-κB修饰(PTM)，包括硝化和磷酸化。这些事件， 与组蛋白脱乙酰酶的募集一起，似乎是抑制SOX18启动子的关键 活动。具体目标#1(SA#1)将确定核因子-κB信号在机械应力调节中的作用 SOX18和CLDN5表达下调，EC连接完整性丧失。SA#1还将调查 ARDS/VILI相关SOX18和CLDN5启动子SNPs对SOX18和CLDN5表达的影响 对机械应力的反应。SA#2将定义硝化介导的PTM在机械应力中的作用- 介导NF-κB信号的激活，并研究机械敏感受体的参与 TRPV4.SA#3将把SAS#1和SAS#2数据转换为可操作的信息，并定义 直接增加SOX18的表达，防止SOX18(HDAC抑制剂)的表观遗传下调， 防止硝化介导的NF-κB激活(屏蔽肽)和减少线粒体来源的ROS (线粒体靶向肽)。连同每个PPG项目和每个PPG中的专业知识 核心，这些探索将促进我们对遗传学、表观遗传学和 PTMS在调节SOX18-CLDN5轴和EC屏障完整性中的作用。此外，我们的研究将促进更多 对VILI病理生物学有透彻的了解，同时强调个体化治疗的应用 病危的人。