SULT4a1, a novel neuroprotective protein in stroke

SULT4a1，一种新型中风神经保护蛋白

• 批准号: 10308473
• 负责人: Shaida A. Andrabi
• 金额: $ 38.48万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10308473
• 关键词: Age; Back; Biological Assay; Biotin; Brain; Cause of Death; Cell Death; Cell Death Signaling Process; Cell Survival; Cells; Cessation of life; Co-Immunoprecipitations; Coagulation Process; Complex; Cytosol; Data; Disease; Drug Metabolic Detoxication; Electron Microscope; Enzymes; Evaluation; Experimental Models; Exposure to; Family; Functional disorder; Gene Targeting; Genus Hippocampus; Glucose; Homeostasis; Hydrogen Peroxide; Image; In Vitro; Inflammatory Response; Knockout Mice; Knowledge; Lead; Ligation; Literature; MG132; Magnetic Resonance Imaging; Mechanics; Mediating; Mediator of activation protein; Middle Cerebral Artery Occlusion; Mitochondria; Modeling; Morbidity - disease rate; Mus; Neurons; Oxidation-Reduction; Oxides; Oxygen; Pathway interactions; Pattern; Peroxides; Pharmacology; Phenotype; Play; Process; Proteins; Proteomics; Regulation; Role; Secondary to; Stains; Stroke; System; TXN gene; Therapeutic Intervention; Thrombectomy; Ubiquitin; Vertebrates; Viral; brain tissue; cell injury; deprivation; disability; in vivo; knock-down; live cell imaging; member; microscopic imaging; mitochondrial dysfunction; neuroprotection; new therapeutic target; novel; novel strategies; peroxiredoxin; peroxiredoxin I; post stroke; relating to nervous system; small hairpin RNA; stroke model; sulfotransferase; targeted treatment; therapeutic target; therapeutically effective; ubiquitin-protein ligase

Stroke remains a leading cause of death and morbidity in the USA and lacks effective therapeutic interventions. Redox imbalance and mitochondrial dysfunction are considered as leading causes of cell death in stroke. Identification of novel therapeutic targets that restore redox homeostasis, mitochondrial function, and cell survival is a critical need. Deregulation in peroxiredoxins (PRDXs) is one of the mechanisms leading to redox imbalance and mitochondrial dysfunction. PRDXs act as double-edged swards that the highly neuroprotective when inside the cells in reduced forms. However, when oxidized and released from damaged/dead cells, PRDXs can lead to secondary cell death signaling via inflammatory responses. Therefore, the regulation of PRDXs in stroke is a vital neuroprotective strategy. We identified that regulation of PRDXs by SULT4a1 (relatively less studies neuronal protein) is a critical neuroprotective function of SULT4a1 and may have a vital neuroprotective role in stroke. Loss of SULT4a1 in mouse neurons leads to the accumulation of oxidized PRDXs with subsequent redox imbalance, mitochondrial dysfunction, and cell death. SULT4a1 can interact with PRDX1, PRDX2, PRDX3, and PRDX5. These PRDXS (cytosolic-PRDX1 and PRDX2, and mitochondrial-PRDX3 and PRDX5) are crucial for cytosolic and mitochondrial redox homeostasis via detoxification of peroxides. During this process, PRDXs are oxidized and must be recycled back to reduced forms. Expression of SULT4a1 increases the levels of reduced PRDXs in peroxide-treated cells. SULT4a1 protein levels decrease in stroke via the ubiquitin proteasomal system. However, the role of SULT4a1 in stroke or the mediators that lead to loss of SULT4a1 in stroke are not known. We will use oxygen- glucose deprivation in mouse cortical neurons and a middle cerebral artery occlusion (MCAO) model of stroke in mice to identify the neuroprotective roles of SULT4a1 in stroke. We propose the following four aims to study the role of SULT4a1 in stroke: Specific Aim #1: Can SULT4a1 maintain redox homeostasis via PRDXs in stroke? Specific Aim #2: Can SULT4a1 rescue mitochondrial function in stroke? Specific Aim #3: Is SULT4a1 neuroprotective in stroke? Specific Aim #4: Identify the mechanisms leading to loss of SULT4a1 in stroke Our supporting data strongly favor our hypothesis and the proposed aims. Combining viral-mediated expression of SULT4a1 with Seahorse Flux analysis, electron microscope imaging, proximity ligation assays, proximity-dependent biotin identification, live-cell imaging, and MRI imaging in experimental models of stroke represents a novel approaches to identify the neuroprotection by SULT4a1 in stroke. These studies will reveal essential knowledge about the role of the novel SULT4a1-dependent pathways in stroke, and determine whether SULT4a1 can represent a credible target for therapeutic intervention in stroke.

中风仍然是美国死亡和发病的主要原因，并且缺乏有效的治疗方法 干预措施。氧化还原失衡和线粒体功能障碍被认为是细胞死亡的主要原因 中风。鉴定恢复氧化还原稳态、线粒体功能、 细胞的生存是一项至关重要的需求。过氧化还原蛋白 (PRDX) 的失调是导致 氧化还原失衡和线粒体功能障碍。 PRDX 是一把双刃剑，高度 当以还原形式存在于细胞内时具有神经保护作用。然而，当氧化并释放时 受损/死亡的细胞，PRDX 可以通过炎症反应导致二次细胞死亡信号。 因此，调节中风中的 PRDX 是一种重要的神经保护策略。我们确定了该法规 SULT4a1（相对较少的研究神经元蛋白）对 PRDX 的影响是关键的神经保护功能 SULT4a1 可能在中风中具有重要的神经保护作用。小鼠神经元中 SULT4a1 的缺失导致 氧化 PRDX 的积累导致氧化还原失衡、线粒体功能障碍和细胞 死亡。 SULT4a1 可以与 PRDX1、PRDX2、PRDX3 和 PRDX5 相互作用。这些 PRDXS（胞质-PRDX1 和 PRDX2、线粒体-PRDX3 和 PRDX5) 对于细胞质和线粒体氧化还原至关重要 通过过氧化物解毒实现体内平衡。在此过程中，PRDX 被氧化，必须回收 回到简化的形式。 SULT4a1 的表达增加了过氧化物处理中还原 PRDX 的水平 细胞。中风时，SULT4a1 蛋白水平通过泛素蛋白酶体系统降低。然而，角色 中风中的 SULT4a1 或导致中风中 SULT4a1 缺失的介质尚不清楚。我们将使用氧气- 小鼠皮质神经元的葡萄糖剥夺和中风的大脑中动脉闭塞（MCAO）模型 在小鼠中鉴定 SULT4a1 在中风中的神经保护作用。我们提出以下四个研究目标 SULT4a1 在中风中的作用： 具体目标 #1：SULT4a1 能否通过 PRDX 在中风中维持氧化还原稳态？ 具体目标 #2：SULT4a1 能否挽救中风患者的线粒体功能？ 具体目标 #3：SULT4a1 对中风有神经保护作用吗？ 具体目标#4：确定导致中风中 SULT4a1 缺失的机制 我们的支持数据强烈支持我们的假设和提出的目标。结合病毒介导 SULT4a1 的表达与 Seahorse Flux 分析、电子显微镜成像、邻近连接测定、 中风实验模型中的邻近依赖性生物素识别、活细胞成像和 MRI 成像 代表了一种识别 SULT4a1 在中风中神经保护作用的新方法。这些研究将揭示 关于新型 SULT4a1 依赖性途径在中风中的作用的基本知识，并确定 SULT4a1 是否可以代表中风治疗干预的可靠靶标。