Oxidative Stress and Pathological Glutamate Release in Stroke

中风中的氧化应激和病理性谷氨酸释放

• 批准号: 10547819
• 负责人: Alexander A Mongin
• 金额: $ 36.45万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10547819
• 关键词: 7-nitroindazole; Acute; Adult; Affect; Alteplase; American; Animals; Anions; Antioxidants; Astrocytes; Basic Science; Behavioral; Biological Assay; Blood - brain barrier anatomy; Blood Preservation; Blood Vessels; Blood flow; Brain; Brain Diseases; Brain Edema; Brain Injuries; Breeding; Cause of Death; Cell Membrane Permeability; Cells; Cerebrovascular Circulation; Chemicals; Clinical; Clinical Trials; Coagulation Process; Complex; Data; Developed Countries; Development; Dextrans; Disabled Persons; Extravasation; FDA approved; Family; Free Radical Scavengers; Genetic; Glutamate Receptor; Glutamates; Goals; Histologic; Human; Injury; Ischemia; Ischemic Brain Injury; Ischemic Penumbra; Japan; Knockout Mice; Lasers; Leucine-Rich Repeat; Link; LoxP-flanked allele; Measures; Mediating; Medicine; Microdialysis; Microfilaments; Middle Cerebral Artery Occlusion; Mission; Mitochondria; Modeling; Molecular; Mus; National Institute of Neurological Disorders and Stroke; Nature; Neuroglia; Neurons; Neurotransmitters; Nitrogen; Oxidants; Oxidation-Reduction; Oxidative Stress; Oxygen; Pathologic; Pathway interactions; Permeability; Pharmaceutical Preparations; Process; Production; Property; Publishing; Rattus; Reactive Oxygen Species; Receptor Activation; Research; Research Personnel; Role; Signal Transduction; Site; Specificity; Stroke; Superoxide Dismutase; Superoxides; Swelling; Tamoxifen; Testing; Therapeutic; Therapeutic Agents; Tissues; United States National Institutes of Health; Viral; Work; acute stroke; behavioral outcome; brain size; channel blockers; clinically relevant; conditional knockout; disability; improved; inhibitor; innovation; ion channel blocker; knock-down; leucine-rich repeat protein; mimetics; neuroprotection; neurovascular; novel therapeutics; overexpression; oxidative damage; pharmacologic; phenylmethylpyrazolone; sex; side effect; stroke model; stroke patient; stroke therapy; tempol; therapeutically effective; tissue injury; tool

SUMMARY Stroke is the fifth leading cause of death and the leading cause of adult long-term disability in the U.S. and other industrialized nations. Yet only one therapeutic agent (the clot-dissolving drug tPA) is approved for acute treatment. Among the critical injurious factors in stroke, oxidative stress is thought to contribute to the terminal steps of tissue damage. Antioxidants and free radical scavengers are highly protective in animal stroke models. Yet, for poorly understood reasons, they have shown limited or no benefits in clinical trials. In the prior NIH- sponsored project, we collected strong data which may help to revise our understanding of the mechanisms contributing to oxidative injury in ischemia: (1) In a rat model of stroke, we found that the superoxide dismutase (SOD) mimetic tempol was more protective than clinically used antioxidants. (2) Potent protection by tempol correlated with reduction of redox-sensitive glutamate release in the ischemic penumbra. (3) Glutamate release was mediated by at least two redox-sensitive mechanisms: volume-regulated anion channels (VRAC) and Ca2+- dependent changes in membrane permeability. (4) The glutamate-permeable VRAC was composed of proteins from the leucine-rich repeat-containing family 8 (LRRC8). We have assembled a synergistic team of investi- gators and propose to use highly innovative molecular and animal tools to test the HYPOTHESIS that reactive oxygen species (ROS, particularly superoxide anions) propagate and amplify stroke injury via stimulation of redox-sensitive glutamate release in the clinically relevant penumbra. In the planned studies we will address the following critical questions: (1) Is the LRRC8A-containing VRAC a viable target for neuroprotection in stroke? (2) Is glutamate release via the heteromeric LRRC8 channels responsible for tissue injury? (3) What is the chemical nature of the tempol-targeted ROS and the cellular site of their production? (4) Does glutamate release via VRAC drive disruption at the neurovascular interface (changes in blood flow and BBB integrity)? The immediate goal of the proposed work is to identify new molecular mechanisms that govern oxidative brain injury and determine the protective actions of antioxidants. Our long-term objective is to provide a blueprint for the development of new effective stroke therapies based on SOD mimetics and/or VRAC blockers.

总结 中风是第五大死亡原因，也是美国和其他国家成人长期残疾的主要原因。 工业化国家。然而，只有一种治疗药物（血栓溶解药物tPA）被批准用于急性 治疗在中风的关键损伤因素中，氧化应激被认为有助于终末 组织损伤的步骤。抗氧化剂和自由基清除剂在动物中风模型中具有高度保护作用。 然而，由于不太清楚的原因，它们在临床试验中显示出有限或没有益处。在之前的NIH- 赞助的项目，我们收集了强有力的数据，这可能有助于修改我们对机制的理解 在缺血性氧化损伤中的作用：（1）在大鼠脑卒中模型中，我们发现超氧化物歧化酶（SOD） (SOD)模拟Tempol比临床使用的抗氧化剂更具有保护作用。(2)tempol的强力保护 与缺血半暗带中氧化还原敏感性谷氨酸释放的减少相关。(3)谷氨酸释放 至少由两种氧化还原敏感机制介导：容量调节阴离子通道（VRAC）和Ca 2 +- 膜通透性的依赖性变化。(4)谷氨酸渗透性VRAC由蛋白质组成 富含亮氨酸的重复序列家族8（LRRC 8）。我们组建了一个协同的投资团队- 并建议使用高度创新的分子和动物工具来测试反应性的假设， 氧物质（ROS，特别是超氧阴离子）通过刺激传播和放大中风损伤 在临床相关半影区的氧化还原敏感性谷氨酸释放。在计划的研究中，我们将 解决以下关键问题：（1）含LRRC 8A的VRAC是否是神经保护的可行靶点 在中风？(2)通过异聚LRRC 8通道释放谷氨酸是组织损伤的原因吗？(3)什么 是tempol靶向ROS的化学性质和它们产生的细胞位点？(4)谷氨酸盐 通过VRAC驱动神经血管界面中断释放（血流和BBB完整性变化）？的 这项工作的近期目标是确定新的控制氧化脑的分子机制 并确定抗氧化剂的保护作用。我们的长远目标是提供一个蓝图， 用于开发基于SOD模拟物和/或VRAC阻断剂的新的有效中风疗法。

项目成果

Conditional deletion of LRRC8A in the brain reduces stroke damage independently of swelling-activated glutamate release.

• DOI: 10.1016/j.isci.2023.106669
• 发表时间: 2023-05-19
• 期刊: ISCIENCE
• 影响因子: 5.8
• 作者: Balkaya, Mustafa;Dohare, Preeti;Chen, Sophie;Schobler, Alexandra L.;Fidaleo, Antonio M.;Nalwalk, Julia W.;Sah, Rajan;Mongin, Alexander A.
• 通讯作者: Mongin, Alexander A.