Redox Control of Seizure-Induced Neuroinflammation

氧化还原控制癫痫引起的神经炎症

• 批准号: 10302913
• 负责人: MANISHA N PATEL
• 金额: $ 2.34万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10302913
• 关键词: Address; Antioxidants; Astrocytes; Automobile Driving; Brain; Cells; Cysteine; Epilepsy; Genetic Transcription; Glial Fibrillary Acidic Protein; Goals; In Vitro; Inflammation; Inflammation Mediators; Ions; Laboratories; Lead; Link; Messenger RNA; Mitochondria; Modification; Mus; Mutate; Neurons; Nutrient; Oxidation-Reduction; Oxidative Stress; Prosencephalon; Proteins; Rattus; Reactive Oxygen Species; Research; Role; SOD2 gene; Seizures; Signal Transduction; Testing; Up-Regulation; Viral Vector; astrogliosis; base; in vivo; mitochondrial dysfunction; neuroinflammation; novel; parent grant; protein expression; therapeutic target

Mitochondrial Reactive Oxygen Species in Epilepsy-Associated Astrogliosis The parent grant focuses on the interplay between neuroinflammation and redox status. The supplemental research addresses a related but distinct goal to investigate the role of mitochondria in seizure-induced neuroinflammation. Mitochondria integrate the energy requirements of neuronal cells and circuits with nutrients, ions, inflammatory mediators and redox status. Mitochondrial dysfunction, oxidative stress and neuroinflammation have been linked with pathophysiological hyperexcitability associated with epilepsy. Neuroinflammation specifically has been identified as a therapeutic target for epilepsy, however the detailed mechanisms underlying seizure-induced neuroinflammation, including upregulation of astrocyte- specific glial fibrillary acidic protein (GFAP), remain at large. One clue regarding upregulation of GFAP arises from recent studies in our laboratory showing a robust transcriptional upregulation of GFAP in mice lacking the anti-oxidant mitochondrial manganese superoxide dismutase-2 (Sod2) in forebrain neurons, a finding replicated in primary neuronal-glial culture. In contrast, mixed cultures in which neurons were selectively depleted displayed no such upregulation. The goal of this project is to determine if mitochondrial reactive oxygen species (mtROS) generated within neurons can activate neuroinflammation via upregulation of GFAP expression as a result of posttranslational redox modification of a conserved cysteine (Cys294) in GFAP, resulting in long-lasting neuroinflammation. Aim 1 will determine if neuronal mtROS is sufficient to induce GFAP upregulation and astrogliosis in vitro using mixed rat neuronal culture and in vivo using Nex-Cre/Sod2f/f mice microinjected with viral vectors driving GCaMP6f within astrocytes. Aim 2 will determine if this astrogliosis involves redox modification of GFAP protein by mutating Cys294. These studies will reveal novel redox-based therapeutic targets to treat epilepsy.

癫痫相关星形胶质细胞增多症中的线粒体活性氧 家长资助的重点是神经炎症和氧化还原状态之间的相互作用。这 补充研究解决了一个相关但独特的目标，即调查 线粒体在癫痫发作引起的神经炎症中的作用。线粒体整合能量 神经元细胞和回路对营养素、离子、炎症介质和 氧化还原状态。线粒体功能障碍、氧化应激和神经炎症 与癫痫相关的病理生理性过度兴奋有关。神经炎症 已被明确确定为癫痫的治疗靶点，但详细信息 癫痫发作引起的神经炎症的机制，包括星形胶质细胞的上调 特定的胶质纤维酸性蛋白（GFAP）仍然逍遥法外。关于上调的一条线索 GFAP 来自我们实验室最近的研究，显示出强大的转录上调 缺乏抗氧化线粒体锰超氧化物歧化酶 2 的小鼠体内 GFAP 的变化 (Sod2) 存在于前脑神经元中，这一发现在原代神经元-胶质细胞培养物中得到了重复。相比之下， 神经元被选择性耗尽的混合培养物没有表现出这种上调。这 该项目的目标是确定线粒体活性氧 (mtROS) 是否产生 神经元内的 GFAP 表达上调，从而激活神经炎症 GFAP 中保守半胱氨酸 (Cys294) 的翻译后氧化还原修饰，导致 长期持续的神经炎症。目标 1 将确定神经元 mtROS 是否足以诱导 使用混合大鼠神经元培养物进行体外 GFAP 上调和星形胶质细胞增生，使用体内使用 Nex-Cre/Sod2f/f 小鼠显微注射了在星形胶质细胞内驱动 GCaMP6f 的病毒载体。目标2 将确定这种星形胶质细胞增生是否涉及通过突变对 GFAP 蛋白进行氧化还原修饰 半胱氨酸294。这些研究将揭示新的基于氧化还原的治疗靶点来治疗癫痫。