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: 8259062
• 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/8259062
• 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）： 这项研究的目的是确定脂质过氧化物诱导的氧化应激对参与肌萎缩性侧面硬化症发病机理（ALS）发病机理的单个组织的影响。许多研究表明，ALS氧化应激具有重要作用。但是，尚未阐明特定类型的氧化损伤和重要的ALS靶细胞。这些信息可能有助于设计对该疾病的急需有效的治疗干预措施。在此提案中，我们将检验以下假设：脂质过氧化物的增加在ALS的起始和/或进展中起关键作用，并且疾病中涉及的细胞类型可能受到差异影响。为此，我们将使用在实验室中开发的小鼠模型来操纵脂质氢过氧化物的水平，这是一种有条件的敲除小鼠，载有磷脂氢过氧化物谷胱甘肽过氧化物酶（GPX4FLFL小鼠）。 GPX4在通过去除脂质氢过氧化物来保护膜中起着至关重要的作用。我们将通过将GPX4FLFL越过携带G93AGUR1 ALS突变体Sod1转基因的小鼠来衡量脂质过氧化物增加对疾病起始和进展的影响。通过在GPX4FLFL条件敲除ALS突变小鼠中的特定组织（例如运动神经元，骨骼肌，小胶质细胞和星形胶质细胞）的靶向表达，我们将产生GPX4的组织特异性缺乏，这是一种关键的抗氧化剂氧化剂氧化剂和型号的氧化型氧化氧化物，并构成了氧化剂的氧化剂。我们将测量疾病发作（体重和神经系统评分的降低），进展（Rotarod，运动神经元数，肌肉神经的丧失）和野生型对照小鼠的寿命，G93A1GUR转基因小鼠，GPX4FLFL FRENSEIC，GPX4FLFL FLLFL小鼠X组织特定的CRE小鼠和G93A1GUR GPX4FL X Telmute X Telmuite Cre tistue Cre cre cyce。我们还将测量对脊髓和胃内胃中胃中胃的蛋白质（碳纤维杆菌），DNA（Oxo8DG）和蛋白质（碳碳，HNE加合物）的氧化损伤，在疾病前，发作和结束阶段，在疾病的疾病前和结束阶段，在疾病对氧化压力上的影响和进展的影响。我们提出的研究将增加我们对一般氧化应激增加的影响，尤其是脂质过氧化物的影响，尤其是在可能与神经退行性疾病ALS的起始和进展有关的特定组织/细胞中。这种测量特定类型的氧化损伤组织特异性影响的新方法具有产生信息的重要潜力，这些信息将不仅可以促进ALS的有效治疗，而且还可以促进涉及运动神经元变性的其他疾病和其他疾病。 公共卫生相关性： ALS是一种令人衰弱的神经退行性疾病，目前尚无有效治愈。对于VA来说，ALS是一个特别重要的问题，因为最近的研究表明，相对于其他人口，海湾战争退伍军人中ALS的发病率显着高于预期。缺乏对ALS患者的有效治疗，再加上该患者人群进行临床试验的固有困难，强调了定义病理生理机制并鉴定潜在治疗靶标的至关重要的重要性。在ALS的特定组织中建立氧化应激的病因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，具有涉及脊髓变性和肌肉萎缩的其他疾病的常见病理。制定靶向疾病进展的干预措施将对涉及脊髓损伤或神经肌肉变性的疾病的治疗具有重要的跨界益处。