Mouse Models of Alexander Disease

亚历山大病小鼠模型

• 批准号: 7877765
• 负责人: ALBEE MESSING
• 金额: $ 36.77万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7877765
• 关键词: Adolescent; Adult; Affect; Alexander Disease; Animal Model; Antioxidants; Astrocytes; Behavioral; Binding; Biochemical; Blood Vessels; Cells; Child; Code; Collaborations; Crystallins; Deposition; Disease; Fiber; Functional disorder; Genes; Genetic; Glial Fibrillary Acidic Protein; Goals; Health; Heat shock proteins; Human; Intermediate Filaments; Knock-in Mouse; Knock-out; Mediating; Modeling; Mus; Mutation; Pathogenesis; Pathologic; Pathway interactions; Patients; Phenotype; Point Mutation; Production; Property; Proteins; Reporter Genes; Response Elements; Role; Specificity; Stress; Testing; Therapeutic Intervention; Transgenic Organisms; Ventricular; base; biological adaptation to stress; gain of function; in vitro Model; in vivo; leukodystrophy; mouse model; mutant; novel; programs; stress protein; transcription factor

DESCRIPTION (provided by applicant): Alexander disease (AxD) patients carry heterozygous mutations within the coding region of GFAP. To facilitate mechanistic studies on the pathogenesis of AxD, and provide animal models suitable for testing potential therapies, our long-term goal has been to generate mouse models for this disorder. We have now generated knock-in lines of mice carrying several of the most common GFAP mu-tations found in human AxD (equivalent to R79H, R239H, R239C, and R416W), and found that expression of the mutant GFAPs induces Rosenthal fibers (the hallmark pathologic feature), but the mice are viable. Moreover, expressing the mutant GFAPs in the context of appropriate genetic modifiers (such as elevating wild type GFAP) results in a lethal phenotype. In addition, altering GFAP expression either by simple over-expression or production of mutant GFAPs leads to induction of multiple stress pathways (aB-crystallin, Nrf2) that suggest specific hypotheses about pathogenesis and, ultimately, strategies for therapy. With the goals of understanding more precisely the consequences of abnormal GFAP expression in astrocytes, and of generating mouse models that reflect key phenotypic features of AxD in humans, we propose the following specific aims: In Aim 1 we will test the consequences of expressing mutant GFAP protein and/or GFAP over-expression, for comparison with known features of the Alexander phenotype in humans. These studies will include both in vitro models using primary cultures of mouse astrocytes, as well as the in vivo knock-in models carrying point mutations in the endogenous mouse GFAP gene. A range of properties will be examined, including biochemical, functional, and behavioral. Reversibility will also be assessed. In Aims 2 and 3 we will test the roles of the small stress protein aB-crystallin, and the transcription factor Nrf2, respectively, as modifiers, by crossing the GFAP mice with knockouts or with newly generated transgenics that over-express the two genes. Specific mechanisms by which excess aB-crystallin or Nrf2 might achieve rescue will be evaluated. These studies promise novel information on the pathological significance of mutant intermediate filament expression in astrocytes, will suggest mechanisms by which primary astrocyte dysfunction leads to generalized CMS disease, and will identify critical stress pathways that could ultimately serve as the basis for therapeutic interventions to mitigate the devastating effects of this disease.

描述（由申请人提供）：亚历山大疾病（AXD）患者在GFAP的编码区域内携带杂合突变。为了促进有关AXD发病机理的机理研究，并提供适合测试潜在疗法的动物模型，我们的长期目标是为这种疾病生成小鼠模型。现在，我们已经产生了携带人AXD中最常见的GFAP MU-tations的小鼠的敲门线（相当于R79H，R239H，R239C和R416W），发现突变GFAP的表达会诱发玫瑰植物纤维（Hallmark Pathologic特征）（Hallmark Pathologic特征），但小鼠的表达均匀。此外，在适当的遗传修饰剂（例如升高野生型GFAP）的背景下表达突变GFAP会导致致命的表型。此外，通过简单的过表达或突变GFAP的产生改变GFAP表达会导致诱导多种应力途径（AB-Crystallin，NRF2），这些途径提出了有关发病机理的特定假设，并最终提出了治疗策略。更精确地理解星形胶质细胞中GFAP表达异常的后果，以及产生反映人类AXD关键表型特征的鼠标模型的后果，我们提出了以下特定目的：在AIM 1中，我们将测试表达突变的GFAP GFAP蛋白质和/或GFAP过度表达的后果，以与Alexandy insexpysement insexpysempys相比。这些研究将包括使用小鼠星形胶质细胞的一级培养物以及内源性小鼠GFAP基因中带有点突变的体内敲入模型的体外模型。将检查一系列属性，包括生化，功能和行为。还将评估可逆性。在AIM 2和3中，我们将通过将GFAP小鼠与敲除或新产生的转基因跨越两个基因的新产生，分别测试小应力蛋白AB-晶状蛋白和转录因子NRF2的作用。将评估过多的AB晶状体或NRF2实现救援的特定机制。这些研究向星形胶质细胞中突变体中间丝表达的病理意义提供了新的信息，将提出机制，通过这些机制，主要的星形胶质细胞功能障碍会导致普遍的CMS疾病，并将确定最终可以作为治疗干预措施的临界应力途径，以减轻这种疾病的灾难性影响。