mitoNEET as a therapeutic target for mitigating ischemic brain injury following MCAO

mitoNEET 作为减轻 MCAO 后缺血性脑损伤的治疗靶点

• 批准号: 10735923
• 负责人: Werner Geldenhuys
• 金额: $ 54.95万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735923
• 关键词: Acute; Address; Advanced Development; Apoproteins; Apoptosis; Area; Attenuated; Basic Science; Bioenergetics; Blood - brain barrier anatomy; Blood brain barrier dysfunction; Brain; Brain Injuries; Cardiovascular system; Cause of Death; Cell Respiration; Cells; Cellular Stress; Clinical Research; Communication; Consumption; Data; Disabled Persons; Disease; Endothelial Cells; Female; Foundations; Generations; Glucose; Goals; Health; In Vitro; Infarction; Iron; Ischemia; Ischemic Brain Injury; Ischemic Stroke; Knowledge; Libraries; Ligands; Link; Lipid Peroxidation; Literature; Mediating; Medical; Metabolic; Middle Cerebral Artery Occlusion; Mission; Mitochondria; Mitochondrial Proteins; Modeling; Mus; National Institute of Neurological Disorders and Stroke; Nervous System Trauma; Neurologic; Neurons; Oncogenic; Outer Mitochondrial Membrane; Oxidation-Reduction; Oxidative Stress; Oxygen; Parents; Pathway interactions; Phase; Pioglitazone; Play; Pre-Clinical Model; Proteins; Public Health; Rattus; Reactive Oxygen Species; Reperfusion Injury; Reperfusion Therapy; Research; Risk; Rodent; Role; Severities; Specificity; Stroke; Sulfur; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Tissues; United States National Institutes of Health; Work; aged; blood-brain barrier disruption; blood-brain barrier function; blood-brain barrier permeabilization; brain endothelial cell; brain tissue; cell injury; cerebral microvasculature; deprivation; design; disability; drug discovery; functional disability; functional improvement; improved; improved outcome; in vitro Model; injured; innovation; insight; interest; male; mitochondrial dysfunction; nervous system disorder; neuroinflammation; neuron loss; neuronal survival; neuropathology; neuroprotection; new therapeutic target; novel; novel therapeutics; post stroke; pre-clinical; preclinical study; repaired; response; sensor; stressor; stroke outcome; stroke recovery; targeted treatment; temporal measurement; therapeutic target; tool

PROJECT SUMMARY Accumulating evidence suggests that following ischemic stroke hypoperfused brain tissue is functionally disabled as electrical communication among penumbral neurons is disrupted due to marked reductions in oxidative metabolism. Thus, it is apparent that mitochondrial dysfunction plays a central role in the degree of neuronal cell death encountered following ischemic brain injury; however, mitochondria have been slow to be fully investigated. Through a serendipitous discovery of an off-target therapeutic effect of pioglitazone, a small mitochondrial iron-sulfur cluster protein, mitoNEET (mNT) was identified that has renewed interest in therapeutic targeting of mitochondria. MitoNEET is embedded in the outer mitochondrial membrane and acts as a redox and pH sensor to regulate mitochondrial bioenergetics, especially in response to cellular stress. Using pioglitazone as our parent compound, we designed NL-1, a first-in-class ligand with high specificity for mN. Using NL-1, we have demonstrated marked improvements in stroke neuropathology and functional impairment following transient middle cerebral artery occlusion (MCAO) in mice and rats. The objective of this proposal is to address fundamental gaps in knowledge regarding how mNT works within the brain to mitigate acute ischemic brain injury. Our central hypothesis is that modulation of mNT acts to improve vulnerable neurons within the penumbra by reducing the vicious cycle of excessive iron-induced lipid peroxidation and increased neuronal death. Based on a strong body of prior literature and pilot data, we postulate the initial target of activity for NL-1 is the cerebral microvasculature; thus, we propose two specific aims to test our hypothesis. In specific aim 1, we will test if mNT selective ligand, NL-1, mitigates ferroptosis using a 4 cell Transwell in vitro model of the blood- brain barrier following oxygen-glucose deprivation with reperfusion. Whereas, in specific aim 2, we will test if mNT ligand, NL-1, reduces brain iron accumulation and blood-brain barrier dysfunction post-MCAO. Successful completion of the proposed research is expected to provide a: (1) greater understanding of how & where mNT mitigates brain injury following ischemic stroke; (2) new insight into the impact of mitochondrial dysfunction & diminished bioenergetics on ischemic stroke outcomes; & (3) strong scientific foundation for an interventional therapeutic approach for treating ischemic stroke. The mechanistic & preclinical data obtained through these studies will serve as critical milestones for advancing the development of mitochondria-targeted therapies for treating neurological injuries & disease.

项目概要 越来越多的证据表明，缺血性中风后，灌注不足的脑组织会出现功能障碍 由于氧化显着减少，半暗神经元之间的电通讯被破坏 代谢。因此，很明显，线粒体功能障碍在神经元细胞的程度中起着核心作用。 缺血性脑损伤后死亡；然而，线粒体要完全发挥作用却很缓慢。 调查了。通过偶然发现吡格列酮（一种小药物）的脱靶治疗作用 线粒体铁硫簇蛋白 mitoNEET (mNT) 的发现重新引起了人们对治疗的兴趣 线粒体靶向。 MitoNEET 嵌入线粒体外膜，充当氧化还原和 pH 传感器调节线粒体生物能量，特别是响应细胞应激。使用吡格列酮 作为我们的母体化合物，我们设计了 NL-1，一种对 mN 具有高特异性的一流配体。使用 NL-1，我们 已证明中风神经病理学和功能障碍有显着改善 小鼠和大鼠的短暂性大脑中动脉闭塞（MCAO）。该提案的目的是解决 关于 mNT 如何在大脑内发挥作用以减轻急性脑缺血的知识存在根本差距 受伤。我们的中心假设是，mNT 的调节可以改善大脑中脆弱的神经元。 通过减少过量铁诱导的脂质过氧化和增加神经元的恶性循环来消除半暗带 死亡。基于大量的先前文献和试点数据，我们假设了 NL-1 的初始活动目标 是脑微血管系统；因此，我们提出两个具体目标来检验我们的假设。在具体目标1中， 我们将使用 4 细胞 Transwell 体外血液模型来测试 mNT 选择性配体 NL-1 是否能减轻铁死亡 氧糖剥夺再灌注后的脑屏障。然而，在具体目标 2 中，我们将测试是否 mNT 配体 NL-1 可减少 MCAO 后的脑铁积累和血脑屏障功能障碍。成功的 完成拟议的研究预计将提供：（1）更好地了解 mNT 的方式和地点 减轻缺血性中风后的脑损伤； (2) 对线粒体功能障碍影响的新见解& 缺血性中风结果的生物能减弱； & (3) 介入治疗的坚实科学基础 治疗缺血性中风的治疗方法。通过这些获得的机械和临床前数据 研究将成为推进线粒体靶向疗法开发的关键里程碑 治疗神经损伤和疾病。