Antioxidants regulators in VALI

VALI 中的抗氧化剂调节剂

• 批准号: 7548530
• 负责人: Sekhar P. Reddy
• 金额: $ 42.71万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7548530
• 关键词: Acute Lung Injury; Antioxidants; Canis familiaris; Cells; Clinical Research; Collaborations; Complex; Development; Enzymes; Epithelial Cells; Event; Family member; Gene Proteins; Genetic; Genomics; Human; Hyperoxia; Injury; Knock-out; Knockout Mice; Lung; Mechanical ventilation; Molecular; Molecular Genetics; Molecular Profiling; Molecular Target; Mus; NF-E2-related factor 2; Oxidants; Oxidative Stress; Phase; Play; Production; Protein Overexpression; Proteins; Proteomics; Reactive Oxygen Species; Research Project Grants; Role; Stress; Stretching; System; Techniques; Testing; Transcription Factor AP-1; Transgenic Mice; Up-Regulation; Validation; Ventilator; base; fos-related antigen 1; insight; lung injury; mutant; protein expression; repaired; response; transcription factor

The overall aim of this SCCOR application is to understand the complex events related to ventilator-associated lung injury (VALI). In Project 6, we will determine the role of oxidative stress in VALI. Oxidative stress has been generally implicated in acute lung injury through production of reactive oxygen species (ROS). To detoxify ROS, lung cells express a network of integrated antioxidant enzymes (AOE) consisting of both classical and phase II detoxifying enzymes. However, if the oxidant stress is overwhelming, ROS may suppress or inactivate the AOE system resulting in cellular damage. We have recently shown by genetic and functional analysis that NF-E2 related factor 2 (Nrf2) plays a critical role in hyperoxia-induced lung injury. This transcription factor is involved in lung epithelial cell repair and stretch and regulates the induction of phase II AOEs. We have also shown that Nrf2-knockout mice are more susceptible to hyperoxia-induced lung injury. In collaboration with the various Cores of the SCCOR, genomic analysis revealed an upregulation of Nrf2 and AP-1 family members, such as fos-related protein-1 (Fra-1), as well as specific AOEs in the lungs of mice and dogs exposed to mechanical ventilation (MV). Thus, we hypothesize that activation of transcription factors such as Nrf2 and Fra-1 and their downstream effectors plays a critical role in the modulation of VALI. To test this hypothesis, we propose the following Specific Aims: 1) Determine the molecular responses of the lung in response to MV and hyperoxia in the development of VALI. We will compare gene and protein expression profiles and markers of injury in the lungs of mice and dogs exposed to MV and hyperoxia; 2) Examine the role of Nrf2 and identify its downstream effectors in VALI; and 3) Examine the role of Fra-1 and identify its downstream effectors in VALI. For these studies, we will use respective knockout and/or transgenic mice with specific lung overexpression of mutant or wildtype proteins of Nrf2 and Fra-1, as well as genomics and proteomics techniques in collaboration with the various Cores. Cross species comparison and validation of these results in human will be done through collaboration with other Basic and Clinical Research Projects, respectively. Results of these studies will provide further insight into the role of oxidative stress as well as the molecular and genetic basis of VALI. The identification of molecular targets will aid in the development of new strategies aimed at minimizing the potential I hazardous effects of MV and hyperoxia.

此 SCCOR 应用的总体目标是了解与呼吸机相关肺损伤 (VALI) 相关的复杂事件。在项目 6 中，我们将确定氧化应激在 VALI 中的作用。氧化应激通常与活性氧 (ROS) 产生引起的急性肺损伤有关。为了解毒 ROS，肺细胞表达一个由经典解毒酶和 II 期解毒酶组成的整合抗氧化酶 (AOE) 网络。然而，如果氧化应激势不可挡，ROS 可能会抑制或灭活 AOE 系统，从而导致细胞损伤。我们最近通过遗传和 功能分析表明NF-E2相关因子2（Nrf2）在高氧诱导的肺损伤中发挥着关键作用。该转录因子参与肺上皮细胞修复和拉伸，并调节 II 期 AOE 的诱导。我们还表明，Nrf2 敲除小鼠更容易受到高氧引起的肺损伤。与 SCCOR 的各个核心合作，基因组分析揭示了 Nrf2 和 AP-1 家族成员的上调，例如 fos 相关蛋白 1 (Fra-1)，以及小鼠和狗肺部的特定 AOE暴露于机械通气（MV）。因此，我们假设 Nrf2 和 Fra-1 等转录因子及其下游效应子的激活在 VALI 的调节中发挥着关键作用。为了检验这一假设，我们提出以下具体目标：1）确定肺在 VALI 发展过程中对 MV 和高氧的分子反应。我们将比较小鼠肺部的基因和蛋白质表达谱以及损伤标志物， 暴露于MV和高氧环境的狗； 2）考察Nrf2在VALI中的作用并鉴定其下游效应子； 3) 检查 Fra-1 的作用并确定其在 VALI 中的下游效应器。对于这些研究，我们将使用具有特定肺过表达 Nrf2 和 Fra-1 突变型或野生型蛋白的相应敲除和/或转基因小鼠，以及与各个 Core 合作的基因组学和蛋白质组学技术。这些结果在人类中的跨物种比较和验证将分别通过与其他基础和临床研究项目的合作来完成。这些研究的结果将进一步深入了解氧化应激的作用以及 VALI 的分子和遗传基础。鉴定 分子靶标的研究将有助于制定新策略，旨在最大限度地减少 MV 和高氧的潜在危险影响。