Glucocerebrosidase mutations in a mouse synucleinopathy model

小鼠突触核蛋白病模型中的葡萄糖脑苷脂酶突变

• 批准号: 7777864
• 负责人: ROBERT L NUSSBAUM
• 金额: $ 13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7777864
• 关键词: Accounting; Affect; Alleles; Alzheimer' s Disease; Animals; Anterior Horn Cells; Artificial Chromosomes; Auditory; Autonomic Dysfunction; Autonomic nervous system; Autopsy; Brain; Brain Diseases; Brain region; Breeding; Catecholamines; Characteristics; Corpus striatum structure; Cultured Cells; Cytoplasmic Protein; Defect; Dementia; Diffuse; Disease; Disease Progression; Disease susceptibility; Elderly; Enzymes; Etiology; Family; Frequencies; Functional disorder; Gastrointestinal Motility; Gaucher Disease; Gene Mutation; Gene Proteins; Genes; Genetic; Genetic Research; Goals; Half-Life; Human; Human Genetics; Infection; Injection of therapeutic agent; Intervention; Knockout Mice; Lewy Bodies; Lewy Body Disease; Lysosomal Storage Diseases; Mental Depression; Missense Mutation; Modeling; Motor; Movement Disorders; Mus; Mutation; Nerve Degeneration; Nervous System Part; Neuraxis; Neurodegenerative Disorders; Neurons; Odds Ratio; Oocytes; Parkinson Disease; Parkinsonian Disorders; Patients; Pattern; Phase; Phenotype; Phosphoproteins; Predisposition; Process; Proteins; Psychotic Disorders; RNA Interference; Reactive Oxygen Species; Regulatory Element; Research; Risk Factors; Rodent Model; Rotenone; SNCA gene; Signs and Symptoms; Stress; Subfamily lentivirinae; Substantia nigra structure; Technology; Testing; Transgenes; Transgenic Mice; Transgenic Organisms; United States; Virus; Visual Hallucination; alpha synuclein; base; cell motility; disability; dopaminergic neuron; dorsal motor nucleus; free radical oxygen; gain of function; gastrointestinal; glucosylceramidase; insight; knock-down; locus ceruleus structure; loss of function; mouse model; mutant; neuropathology; olfactory bulb; overexpression; presynaptic; promoter; protein aggregate; public health relevance; response; small hairpin RNA; synuclein

DESCRIPTION (provided by applicant): Parkinson Disease (PD) is a common, mostly sporadic neurodegenerative disease of the central and autonomic nervous systems affecting ~1 million people in the United States. Loss of substantia nigra (SN) dopaminergic neurons causes the characteristic parkinsonian movement disorder while non-motor disabilities including dementia, depression, gastrointestinal dysfunction, and autonomic instability arise from neurodegeneration in other parts of the nervous system. The cytoplasmic protein aggregates known as Lewy bodies, described nearly 100 years ago in the regions of the brain affected by PD, such as the substantia nigra, locus coeruleus, olfactory bulbs, and dorsal motor nucleus of the vagus, are considered to be the pathological hallmark of the disease. Diffuse Lewy body dementia (DLBD) accounts for ~10% of cases of all dementia, although some estimates suggest it is nearly as frequent as Alzheimer disease (AD) as a cause of dementia in the elderly. DLBD is clinically similar to AD but characteristically shows a higher frequency of psychosis with auditory and visual hallucinations and is often accompanied by the symptoms and signs of Parkinsonism. At autopsy, brains of DLBD patients have characteristic Lewy body aggregates throughout the cortex. The primary protein constituent of Lewy bodies is a-synuclein (aSyn), a 140 residue presynaptic phosphoprotein. Missense and increased copy number mutations in the gene for aSyn cause familial, autosomal dominant PD and DLBD. Since (1) aSyn-containing Lewy bodies are seen in both DLBD and PD, and (2) many of the patients in families with increased copy number mutations of aSyn have DLBD while others have PD11-15, it is very likely that the same underlying process is responsible for both conditions. The cause of most PD is unknown despite decades of research. Recently, a strong epidemological association has been found between PD/DLBD and heterozygosity for mutations in the beta-glucocerebrosidase gene (GBA), the gene in which homozygous mutations cause Gaucher disease (GD), a lysosomal storage disease. The odds ratio of finding a heterozygous GBA mutation in sporadic PD patients compared to controls ranges from the 13-28 depending on ethnic background. Thus, GBA mutations are a clear risk factor for PD but the mechanism is unknown. Being heterozygous for these mutations could increase susceptibility for PD either through haploinsufficiency of lysosomal glucocerebrosidase or through a toxic gain of function by the mutant gene product encoded by the mutant allele. The overall goal of this proposal is to develop a mouse model that uses expression of a mutant a-synuclein known to cause familial PD/DLBD to which we add either loss of function of glucocerebrosidase or expression of a mutant form of glucocerebrosidase to test for (1) changes in aSyn half-life and sensitivity to oxygen free radical challenge in cultured cells, and (2) motor and non-motor signs of PD in mice carrying pathogenic aSyn alleles and loss of function of glucocerebrosidase or expression of a mutant form of glucocerebrosidase. PUBLIC HEALTH RELEVANCE: Parkinson Disease and diffuse Lewy body dementia are common degenerative brain diseases affecting ~1 million people in the United States. The applicant wants to know if an authentic mouse model of these diseases can be created by combining genetic factors in a single mouse. These factors are the overexpression of a mutant form of 1-synuclein known to cause Parkinson disease in humans along with changes in the expression of the enzyme glucocerebrosidase, a Parkinson disease susceptibility risk factor. The ultimate goal is to create a mouse line that can be widely used to study in detail how familial Parkinson disease develops in order to identify points at which interventions could slow or stop progression of the disease.

描述（由申请人提供）：帕金森病（PD）是一种常见的、大多数为散发性的中枢和自主神经系统神经退行性疾病，影响美国约100万人。黑质（SN）多巴胺能神经元的损失引起特征性帕金森运动障碍，而非运动残疾包括痴呆、抑郁、胃肠功能障碍和自主神经不稳定由神经系统的其他部分中的神经变性引起。近100年前在受PD影响的大脑区域（如黑质、蓝斑、嗅球和迷走神经背侧运动核）中描述的称为路易体的细胞质蛋白聚集体被认为是该疾病的病理标志。弥漫性路易体痴呆（DLBD）占所有痴呆病例的约10%，尽管一些估计表明它几乎与阿尔茨海默病（AD）一样常见，是老年痴呆的原因。DLBD在临床上类似于AD，但特征性地显示出更高频率的精神病，伴有听觉和视觉幻觉，并且通常伴有帕金森综合征的症状和体征。在尸检中，DLBD患者的大脑在整个皮质中具有特征性的路易体聚集体。路易体的主要蛋白质成分是α-突触核蛋白（aSyn），一种140个残基的突触前磷蛋白。aSyn基因中的错义和拷贝数增加突变导致家族性常染色体显性PD和DLBD。由于（1）在DLBD和PD中均观察到含有aSyn的路易体，以及（2）aSyn拷贝数突变增加的家族中的许多患者患有DLBD，而其他患者患有PD 11 -15，因此很可能相同的潜在过程是导致这两种病症的原因。大多数PD的原因是未知的，尽管几十年的研究。最近，在PD/DLBD和β-葡糖脑苷脂酶基因（GBA）突变的杂合性之间发现了强烈的药理学关联，该基因中纯合突变导致戈谢病（GD），一种溶酶体贮积病。与对照组相比，在散发性PD患者中发现杂合GBA突变的比值比范围为13-28，具体取决于种族背景。因此，GBA突变是PD的明确风险因素，但机制尚不清楚。这些突变的杂合子可能通过溶酶体葡萄糖脑苷脂酶的单倍不足或通过突变等位基因编码的突变基因产物的毒性功能获得而增加PD的易感性。该提议的总体目标是开发一种小鼠模型，其使用已知引起家族性PD/DLBD的突变体α-突触核蛋白的表达，我们向其添加葡糖脑苷脂酶的功能丧失或葡糖脑苷脂酶的突变体形式的表达，以测试（1）培养细胞中aSyn半衰期和对氧自由基攻击的敏感性的变化，和（2）携带致病性aSyn等位基因和葡糖脑苷脂酶功能丧失或葡糖脑苷脂酶突变形式表达的小鼠中PD的运动和非运动体征。公共卫生相关性：帕金森病和弥漫性路易体痴呆是常见的退行性脑疾病，在美国影响约100万人。申请人想知道是否可以通过在单个小鼠中组合遗传因子来创建这些疾病的真实小鼠模型。这些因素是已知引起人类帕金森病的1-突触核蛋白的突变形式的过表达以及葡萄糖脑苷脂酶（帕金森病易感性风险因素）表达的变化沿着。最终目标是创建一个小鼠系，可广泛用于详细研究家族性帕金森病如何发展，以确定干预措施可以减缓或阻止疾病进展的点。