Targeted investigation of tissue specific oxidative stress in the etiology of ALS

ALS 病因中组织特异性氧化应激的针对性研究

• 批准号: 7930157
• 负责人: HOLLY VAN REMMEN
• 金额: --
• 依托单位: SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7930157
• 关键词: Actins; Affect; Alleles; Amyotrophic Lateral Sclerosis; Antioxidants; Astrocytes; Body Weight decreased; Breeding; Cells; Cessation of life; Clinical Trials; Coupled; DNA; Disease; Disease Progression; Enzymes; Etiology; Excision; F2-Isoprostanes; Gastrocnemius Muscle; Genetic Crossing Over; Glial Fibrillary Acidic Protein; Gulf War; Human; ITGAM gene; Incidence; Individual; Intervention; Investigation; Knock-out; Knockout Mice; Laboratories; Lipid Peroxidation; Lipid Peroxides; Lipids; Longevity; Measures; Mediating; Mediator of activation protein; Membrane; Microglia; Motor Neurons; Mus; Muscle; Muscle Weakness; Muscular Atrophy; Mutant Strains Mice; Neurodegenerative Disorders; Neuroglia; Neurologic; Neurons; Onset of illness; Oxidative Stress; Paralysed; Pathogenesis; Pathology; Patients; Play; Population; Proteins; Relative (related person); Rest; Role; Skeletal Muscle; Spinal Cord; Spinal cord injury; Testing; Therapeutic Intervention; Tissues; Transgenes; Transgenic Mice; Veterans; adduct; cell type; design; effective therapy; end stage disease; motor neuron degeneration; mouse model; mutant; nerve supply; neuromuscular; neuron loss; novel strategies; oxidation; oxidative damage; patient population; phospholipid-hydroperoxide glutathione peroxidase; recombinase; response; skeletal; therapeutic target

DESCRIPTION (provided by applicant): The purpose of this study is to determine the impact of lipid peroxide induced oxidative stress on individual tissues involved in the pathogenesis of Amyotrophic Lateral Sclerosis (ALS). Many studies have suggested an important role for oxidative stress in ALS; however, the specific types of oxidative damage and the target cells that are important in ALS have not been elucidated. This information could be instrumental in helping to design much needed effective therapeutic interventions for this disease. In this proposal, we will test the hypothesis that an increase in lipid peroxides plays a critical role in the initiation and/or progression of ALS and that the cell types involved in the disease may be differentially affected. To do this, we will use a mouse model developed in our laboratory to manipulate the levels of lipid hydroperoxides, a conditional knockout mouse carrying floxed alleles for phospholipid hydroperoxide glutathione peroxidase (Gpx4flfl mice). Gpx4 plays a critical role in protecting membranes by removal of lipid hydroperoxides. We will measure the effect of increased lipid peroxides in specific tissues/cell types on disease initiation and progression by crossing the Gpx4flfl to mice carrying the G93AGur1 ALS mutant SOD1 transgene. Through targeted expression of Cre recombinase to specific tissues (e.g., motor neurons, skeletal muscle, microglia and astrocytes) in the Gpx4flfl conditional knockout ALS mutant mice, we will generate tissue-specific deletion of Gpx4, a key antioxidant enzyme, and increase lipid hydroperoxide induced oxidative stress in the targeted tissues/cell types. We will measure disease onset (loss of body weight and neurologic score), progression (rotarod, motor neuron numbers, loss of muscle innervation) and lifespan in wild type control mice, G93A1Gur transgenic mice, Gpx4flfl mice x tissue specific Cre mice and G93A1Gur Gpx4flfl x tissue specific Cre mice. We will also measure oxidative damage to lipid (F2-isoprostanes, neuroprostanes), DNA (oxo8dG) and protein (carbonyls, HNE adducts) in spinal cord and gastrocnemius muscle at pre-symptomatic, onset and end stages of the disease to determine the impact of increased oxidative stress on disease initiation and progression in these tissues. The studies we have proposed will increase our understanding of the effect of increased oxidative stress in general, and lipid peroxides in particular, in specific tissues/cells potentially involved in the initiation and progression of the neurodegenerative disease ALS. This novel approach to measure tissue specific effects of a specific type of oxidative damage has significant potential to generate information that will facilitate the design of effective treatments not only for ALS, but also for and other diseases involving motor neuron degeneration. PUBLIC HEALTH RELEVANCE: ALS is a debilitating neurodegenerative disease that currently has no effective cure. ALS is a particularly important issue for the VA as recent studies have shown that the incidence of ALS among Gulf War Veterans is significantly higher than expected relative to the rest of the population. The lack of sustained effective treatment for patients with ALS, coupled with the inherent difficulty in performing clinical trials in this patient population, highlights the critical importance of defining pathophysiological mechanisms and identifying potential therapeutic targets. Establishing a causative role for oxidative stress in specific tissues in ALS will offer new potential strategies for therapeutic interventions. In addition, ALS is a neurodegenerative disease that has common pathologies with other diseases involving spinal cord degeneration and muscle atrophy. Developing interventions to target disease progression in ALS will have important cross-over benefits for treatment of diseases involving spinal cord injury or neuromuscular degeneration.

描述（由申请人提供）： 本研究的目的是确定脂质过氧化物诱导的氧化应激对肌萎缩侧索硬化（ALS）发病机制中涉及的个体组织的影响。许多研究表明氧化应激在ALS中起重要作用;然而，氧化损伤的具体类型和在ALS中重要的靶细胞尚未阐明。这些信息可能有助于设计急需的有效的治疗干预措施，这种疾病。在这项提案中，我们将测试的假设，脂质过氧化物的增加在ALS的启动和/或进展中起着至关重要的作用，并参与疾病的细胞类型可能会受到不同的影响。为此，我们将使用我们实验室开发的小鼠模型来操纵脂质氢过氧化物的水平，这是一种携带磷脂氢过氧化物谷胱甘肽过氧化物酶floxed等位基因的条件性敲除小鼠（Gpx 4flfl小鼠）。gpx 4通过清除脂质过氧化氢在保护膜中起着关键作用。我们将通过将Gpx 4flfl与携带G93 AGur 1 ALS突变体SOD 1转基因的小鼠杂交来测量特定组织/细胞类型中脂质过氧化物增加对疾病起始和进展的影响。通过将Cre重组酶靶向表达至特定组织（例如，运动神经元、骨骼肌、小神经胶质细胞和星形胶质细胞），我们将产生Gpx 4（一种关键的抗氧化酶）的组织特异性缺失，并在靶组织/细胞类型中增加脂质过氧化氢诱导的氧化应激。我们将测量野生型对照小鼠、G93 A1 Gur转基因小鼠、Gpx 4flfl小鼠x组织特异性Cre小鼠和G93 A1 Gur Gpx 4flfl x组织特异性Cre小鼠中的疾病发作（体重减轻和神经学评分）、进展（转棒、运动神经元数目、肌肉神经支配的丧失）和寿命。我们还将在疾病的症状前、发作和结束阶段测量脊髓和腓肠肌中脂质（F2-异前列烷、神经前列烷）、DNA（oxo 8dG）和蛋白质（羰基、HNE加合物）的氧化损伤，以确定氧化应激增加对这些组织中疾病发生和进展的影响。我们提出的研究将增加我们对一般氧化应激增加的影响的理解，特别是脂质过氧化物，在可能参与神经退行性疾病ALS的起始和进展的特定组织/细胞中。这种测量特定类型氧化损伤的组织特异性效应的新方法具有产生信息的显著潜力，这些信息将有助于设计有效的治疗方法，不仅用于ALS，还用于其他涉及运动神经元变性的疾病。 公共卫生关系： ALS是一种使人衰弱的神经退行性疾病，目前还没有有效的治疗方法。ALS是VA的一个特别重要的问题，因为最近的研究表明，海湾战争退伍军人中ALS的发病率明显高于其他人群的预期。ALS患者缺乏持续有效的治疗，加上在该患者人群中进行临床试验的固有困难，突出了定义病理生理机制和确定潜在治疗靶点的至关重要性。建立氧化应激在ALS特定组织中的致病作用将为治疗干预提供新的潜在策略。此外，ALS是一种神经退行性疾病，与涉及脊髓变性和肌肉萎缩的其他疾病具有共同的病理学。开发针对ALS疾病进展的干预措施将对治疗涉及脊髓损伤或神经肌肉变性的疾病具有重要的交叉益处。