Constructing a Conditional Slc7a11 (xCT) Null Mouse

构建条件 Slc7a11 (xCT) 空鼠标

• 批准号: 8203292
• 负责人: SANDRA J HEWETT
• 金额: $ 14.7万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8203292
• 关键词: Alleles; Amino Acid Transporter; Animal Model; Antioxidants; Astrocytes; Behavioral; Brain; Brain Injuries; Cell Culture Techniques; Cell membrane; Cells; Cerebral Ischemia; Chemosensitization; Couples; Cyst; Cysteine; Cystine; Development; Economic Burden; Emotional; Environment; Enzymes; Excitatory Amino Acid Receptors; Exons; Free Radicals; Gene Expression; Generations; Genes; Genetic Transcription; Glutamate Receptor; Glutamate Transporter; Glutamates; Glutathione; Glycine; Goals; Health; Hour; Hypoxia; Inflammatory; Injury; Interleukin-12; Interleukins; Invaded; Ischemic Penumbra; Knock-out; Knockout Mice; Laboratories; Lead; Light; Literature; Maintenance; Mediating; Metabolism; Microglia; Morbidity - disease rate; Mus; Mutant Strains Mice; Nerve Degeneration; Neuronal Injury; Neurons; Organism; Oxidants; Oxidative Stress; Play; Prevention; Process; Production; Proteins; Public Health; Recycling; Role; Signal Transduction; Site; Stroke; Substrate Specificity; System; Tissues; Transgenic Mice; Translations; antiporter; cell type; cofactor; cytokine; design; driving force; extracellular; glutathione peroxidase; in vitro Model; in vivo; knockout animal; loss of function; loss of function mutation; new therapeutic target; prevent; stoichiometry; uptake

DESCRIPTION (provided by applicant): Injury to the brain caused by cerebral ischemia is a major public health concern. Studies have determined that the brain damage associated with cerebral ischemia is mediated by over-stimulation of excitatory amino acid receptors, oxidative stress, as well as inflammatory factors. Our laboratory demonstrated - using an in vitro model of the ischemic penumbra - that astrocyte-mediated alterations in system xc- (cystine/glutamate antiporter) contribute to the development and progression of inflammatory (IL-12-mediated) hypoxic neuronal injury. Thus, we believe that system xc- has the potential to be a novel therapeutic target for stroke. However, to validate this hypothesis, our results must be confirmed in vivo. Mice harboring a natural loss of function mutation in the Slc7a11 (sut) gene - which encodes for xCT, the light chain dictating substrate specificity in system xc- - and xCT null mutant mice have been described in the literature. However, these are global knockouts. To date no tissue-specific knockout for this allele exists. Hence, the overall goal of this project is to develop (Aim 1) and characterize (Aim 2) an astrocyte-specific conditional knockout mouse of the Slc7a11 gene for the ultimate use in in vivo. PUBLIC HEALTH RELEVANCE: Morbidity associated with cerebral ischemia (stroke) remains a huge emotional and economic burden due in large part to a void in treatment options to protect against secondary injury. It is our contention that successful completion of this application [i.e., the successful generation of a Scla711 (xCT) conditional knockout animal] will allow us to assess the viability of our hypothesis that astrocyte system xc- (cystine/glutamate transporter) is a novel therapeutic target in stroke.

描述（由申请人提供）：脑缺血引起的脑损伤是一个主要的公共卫生问题。研究表明，与脑缺血相关的脑损伤是由兴奋性氨基酸受体的过度刺激、氧化应激以及炎症因子介导的。我们的实验室使用缺血半暗带的体外模型证明，星形胶质细胞介导的 xc 系统（胱氨酸/谷氨酸逆向转运蛋白）的改变有助于炎症（IL-12 介导的）缺氧神经元损伤的发生和进展。因此，我们相信系统xc-有潜力成为中风的新治疗靶点。然而，为了验证这一假设，我们的结果必须在体内得到证实。文献中已经描述了 Slc7a11 (sut) 基因（编码 xCT，决定 xc 系统中底物特异性的轻链）中具有自然功能缺失突变的小鼠和 xCT 无效突变小鼠。然而，这些都是全球淘汰赛。迄今为止，还没有针对该等位基因的组织特异性敲除。因此，该项目的总体目标是开发（目标 1）并表征（目标 2）Slc7a11 基因星形胶质细胞特异性条件敲除小鼠，以最终在体内使用。 公共卫生相关性：与脑缺血（中风）相关的发病率仍然是巨大的情感和经济负担，这在很大程度上是由于缺乏防止继发性损伤的治疗方案。我们认为，成功完成该应用[即成功生成 Scla711 (xCT) 条件敲除动物]将使我们能够评估我们的假设的可行性，即星形胶质细胞系统 xc-（胱氨酸/谷氨酸转运蛋白）是中风的新型治疗靶点。