Role of prohibitin in ischemic brain injury

抑制素在缺血性脑损伤中的作用

• 批准号: 10201370
• 负责人: Ping Zhou
• 金额: $ 49.92万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201370
• 关键词: Address; Affect; Astrocytes; Bioenergetics; Biological Assay; Brain; Brain Injuries; Calcium; Cell Culture Techniques; Cell Survival; Cells; Cellular Assay; Cerebrum; Coculture Techniques; Complement; Complex; Cysteine; Disease; Embryo; Environment; Funding; Gel; Genetic; Genus Hippocampus; Glucose; Ischemia; Ischemic Brain Injury; Ischemic Neuronal Injury; Ischemic Preconditioning; Knock-in; Knock-in Mouse; Knock-out; Knockout Mice; Life; Link; Maintenance; Mediating; Mitochondria; Mitochondrial Proteins; Modeling; Molecular; Molecular Weight; Mouse Protein; Mus; Mutant Strains Mice; Mutate; Mutation; Neuroglia; Neurologic; Neurons; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthetase Inhibitor; Open Reading Frames; Oxidative Phosphorylation; Oxygen; Patients; Play; Process; Production; Protein S; Proteins; Reactive Oxygen Species; Regulation; Respiratory Chain; Role; S-Nitrosoglutathione; S-Nitrosothiols; Somatic Mutation; Stress; Stroke; Subfamily lentivirinae; System; Testing; Transgenic Mice; Up-Regulation; Work; artery occlusion; base; cell type; conditional knockout; deprivation; in vivo; ischemic injury; mitochondrial dysfunction; mutant; nervous system disorder; neuron loss; neuronal survival; neuroprotection; novel; overexpression; prevent; prohibitin; protective effect; stroke risk; targeted treatment; treatment strategy

ABSTRACT The mitochondrial protein Prohibitin (PHB) is essential for life. Its importance to cellular activities is attested by the fact that deletion of PHB is embryonic lethal in mice and that to date no mutation has been found in the coding region of PHB in any disease conditions, indicating that PHB integrity is essential and that somatic mutation is detrimental. PHB has been shown to be critical for mitochondrial function in stress situations, as it is upregulated in ischemic preconditioning (IPC). In the previous funding period, using neuron specific PHB transgenic mice, we demonstrated that selective neuronal PHB expression leads to remarkable neuroprotection against middle cerebral arterial occlusion (MCAO) induced brain injury. However, how this important protein is functionally regulated in IPC, as well as how it is dysregulated in other neurological conditions, remain surprisingly unknown. In exploring the mechanisms of PHB regulation, we discovered that nitric oxide (NO) is required for both IPC and PHB upregulation. Therefore, we investigated the interaction between NO and PHB and found that NO modifies PHB post-translationally, through protein s-nitrosylation. In this renewal application, we propose to study the effects of PHB S- nitrosylation and the mechanisms underlying functional regulation of PHB by NO. Our central hypothesis is that PHB nitrosylation is critical for its neuroprotective function and that disturbances of PHB nitrosylation is detrimental. We will use a mutant knockin mouse, in which the sole cysteine residue of PHB protein is mutated so that PHB cannot be nitrosylated, and a PHB neuronal knockout mouse, in which PHB deletion is complemented by AAV expressing wild type or non-nitrosylated C69S mutant to analyze the mechanisms of NO regulation and the effects of loss of PHB nitrosylation on PHB function, in IPC. Three specific aims will systematically test the hypothesis. The results of the proposed studies will reveal a previously undescribed regulatory mechanism of PHB and could benefit patients at risk of stroke and other neurological conditions.

抽象的 线粒体蛋白抑制素 (PHB) 对于生命至关重要。它对细胞活动的重要性 事实证明，PHB 的缺失对小鼠胚胎是致命的，并且迄今为止没有突变 在任何疾病条件下的 PHB 编码区中都发现了 PHB 的完整性 是必不可少的，并且体细胞突变是有害的。 PHB 已被证明对于 线粒体在应激情况下的功能，因为它在缺血预适应（IPC）中上调。 在之前的资助期间，我们使用神经元特异性 PHB 转基因小鼠证明了 选择性神经元 PHB 表达可对中脑产生显着的神经保护作用 动脉闭塞（MCAO）引起的脑损伤。然而，这种重要的蛋白质如何发挥功能 IPC 中的调节，以及它在其他神经系统疾病中的失调情况，仍然存在 令人惊讶的是未知。在探索 PHB 调节机制时，我们发现硝酸 IPC 和 PHB 上调都需要氧化物 (NO)。因此，我们调查了 NO 和 PHB 之间的相互作用，发现 NO 通过翻译后修饰 PHB 蛋白质 s-亚硝基化。在此更新申请中，我们建议研究 PHB S- 的影响 亚硝基化和 NO 对 PHB 功能调节的潜在机制。我们的中央 假设是 PHB 亚硝基化对其神经保护功能至关重要，并且干扰 PHB 的亚硝基化是有害的。我们将使用突变型敲入小鼠，其中鞋底 PHB蛋白的半胱氨酸残基发生突变，使得PHB不能被亚硝基化，并且PHB 神经元敲除小鼠，其中 PHB 缺失由表达野生型的 AAV 补充 或非亚硝基化 C69S 突变体来分析 NO 调节机制以及 IPC 中 PHB 亚硝基化的丧失对 PHB 功能的影响。三个具体目标将系统地测试 假设。拟议研究的结果将揭示先前未描述的监管 PHB 的机制，可以使有中风和其他神经系统疾病风险的患者受益。