Cell biological analysis of CACH/VWM leukodystrophy

CACH/VWM 脑白质营养不良的细胞生物学分析

• 批准号: 6823914
• 负责人: CHRISTOPH PROSCHEL
• 金额: $ 18.04万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6823914
• 关键词: astrocytes; cell biology; cell population study; cellular pathology; disease /disorder etiology; gene dosage; gene mutation; glial fibrillary acidic protein; human tissue; hyperthermia; leukodystrophy; myelinopathy; oligodendroglia; physiologic stressor; tissue /cell culture

DESCRIPTION (provided by applicant): Diseases of militated white matter tracts in the central nervous system (CNS) represent one of the most frequent neurological disorders. Those known to be inheritable are referred to as leukodystrophies. In many instances the identification of specific genetic lesions has helped explain the pathology observed in patients. This has not been the case for the recently identified mutations in the subunits of translation initiation factor 2B (eIF2B) in patients diagnosed with autosomal recessive Childhood Ataxia with Diffuse CNS Hypomyelination (CACH)/Vanishing White Matter Disease (VWM; MIM 603896). The lack of a suitable experimental system in which to directly test the effect of elF2B mutations on the biology of neural cell populations is a major obstacle in understanding the etiology of CACH/VWM disease. By applying our extensive experience in the isolation and culture of neural stem ceils and progenitors from both the rodent and human CNS, we established cultures of ceils with the characteristics of neural precursors from the brain of a CACH/VWM disease patient with known mutations in the epsilon subunit of elF2B (EIF2B5). This is the first example of an in vitro system using primary ceils directly isolated from a leukodystrophy patient. Analysis of these cultures using lineage specific markers revealed the presence of neurons, oligodendrocytes and astrocytes. In light of the clinical pathology, we initially focused on the oligodendrocyte compartment. Contrary to our expectations, oligodendrocytes derived from the patient's brain appeared normal by morphological criteria and progressive maturation of oligodendroglial lineage cells could be observed. However, only few glial fiballry acidic protein (GFAP) expressing cells were present and most of these ceils exhibited an atypical morphology. Also, the induction of GFAP + astrocytes using otherwise highly pro-astrocytic conditions was severely impaired. This suggests that a mutation in eIF2B5 leads to a defect in the formation of astrocytes in CACH/VWM disease. This does not preclude that oligodendrocytes may also be directly affected. However, the possibility that a deficiency in astrocyte function may contribute to the loss of white matter in CACH/VWM leukodystrophy in a non autonomous manner has important consequences for therapeutic strategies. The goal of the proposed research is to exploit this unique in vitro system to study the biological effects of eiF2B5 mutations on different CNS lineages in CACH/VWM disease patients.

描述（由申请人提供）： 中枢神经系统（CNS）中的受损白色束疾病是最常见的神经系统疾病之一。那些已知是可遗传的被称为脑白质营养不良。在许多情况下，特定遗传病变的鉴定有助于解释在患者中观察到的病理。最近在诊断为常染色体隐性遗传性儿童共济失调伴弥漫性CNS髓鞘形成不足（CACH）/消失性白色物质病（VWM; MIM 603896）的患者中发现的翻译起始因子2B（eIF 2B）亚基突变并非如此。 缺乏合适的实验系统来直接测试eIF 2B突变对神经细胞群体的生物学的影响，这是理解CACH/VWM疾病的病因的主要障碍。通过应用我们在分离和培养来自啮齿动物和人CNS的神经干细胞和祖细胞方面的丰富经验，我们建立了具有来自CACH/VWM疾病患者的脑的神经前体细胞特征的细胞培养物，该患者在eIF 2B的EIF 2B 5亚基中具有已知突变。这是使用直接从脑白质营养不良患者分离的原代细胞的体外系统的第一个实例。 使用谱系特异性标记物对这些培养物的分析揭示了神经元、少突胶质细胞和星形胶质细胞的存在。根据临床病理学，我们最初专注于少突胶质细胞区室。与我们的预期相反，从患者的大脑中提取的少突胶质细胞的形态学标准似乎正常，并且可以观察到少突胶质细胞谱系细胞的逐渐成熟。然而，仅存在少数表达胶质纤维酸性蛋白（GFAP）的细胞，并且大多数这些细胞表现出非典型形态。此外，使用其他高度促星形胶质细胞条件的GFAP +星形胶质细胞的诱导严重受损。这表明eIF 2B 5突变导致CACH/VWM疾病中星形胶质细胞形成缺陷。这并不排除少突胶质细胞也可能直接受到影响。然而，星形胶质细胞功能缺陷可能导致CACH/VWM脑白质营养不良中白色物质以非自主方式丢失的可能性对治疗策略具有重要影响。该研究的目的是利用这种独特的体外系统来研究eiF 2B 5突变对CACH/VWM疾病患者不同CNS谱系的生物学效应。