iPSC-derived astrocytes to model Vanishing White Matter Disease

iPSC 衍生的星形胶质细胞用于模拟白质消失疾病

• 批准号: 9763666
• 负责人: CHRISTOPH PROSCHEL
• 金额: $ 19.99万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9763666
• 关键词: Address; Affect; Astrocytes; Ataxia; Autopsy; Biology; Biopsy Specimen; Blood specimen; Brain; CRISPR/Cas technology; Cell Culture Techniques; Cell Death; Cell Line; Cell Lineage; Cells; Childhood; Clinic; Complex; Cues; Data; Defect; Diffuse; Disease; Drug Screening; EIF2B2 gene; EIF2B5 gene; Etiology; Eukaryotic Initiation Factors; Exhibits; Generations; Genetic; Genotype; Glial Fibrillary Acidic Protein; Gliosis; Goals; Hereditary Disease; Homeostasis; Human; Human Biology; Impairment; In Vitro; Individual; Inflammatory; Lesion; Light; Modeling; Morphology; Mus; Mutation; Myelin; Nerve; Neuraxis; Neurons; Oligodendroglia; Pathology; Patients; Penetrance; Peptide Initiation Factors; Phenotype; Play; Pluripotent Stem Cells; Population; Property; Proteins; Protocols documentation; Resources; Rodent; Role; Sampling; Site; Suggestion; System; Testing; Tissue Sample; United States; White Matter Disease; biomedical referral center; brain cell; cell type; drug development; experience; insight; leukodystrophy; mouse model; mutant; nerve stem cell; nervous system disorder; oligodendrocyte precursor; precursor cell; prevent; protein complex; response; stem cell technology; tool; white matter

Vanishing White Matter (VWM) disease is one of the most prevalent leukodystrophies, and is caused by mutations in any of the five subunits of eukaryotic translation initiation factor EIF2B. Even though many different kinds of EIF2B mutations have been identified, it is still unclear how these mutations cause the dramatic loss of myelin. One of the biggest problems has been the lack of a suitable experimental system in which to test the effects of EIF2B mutations on brain cells. Our initial study of VWM patient-derived cells was made possible by the exceedingly rare opportunity to isolate live glial precursor cells from the brain of a VWM patient. This study revealed a defect in astrocytes. But the lack of suitable patient samples limits our ability to study the cellular causes and consequently prevent us from finding a treatment for this devastating disease. It is the goal of this proposal to exploit inducible pluripotent stem cell (iPSC) technology to derive neural stem cell cultures from VWM-patient with distinct EIF2B complex mutations. These VWM-neural stem cells will be used to test the genotype-phenotype relationship with respect to astrocyte differentiation and function. Our preliminary data demonstrate that iPSCs can readily be derived from VWM-patient cells and that these patient- derived PSCs can be induced to differentiate into neural stem cells and astrocyte precursors. Mutations in EIF2B appear to cause increased cell death, and impair expression of some astroglial lineage proteins, including the astroglial lineage determinant CREB3L1. Because of the critical role of astrocytes in maintaining CNS homeostasis, our findings raise two important questions: (1) Do VWM patients with distinct EIF2B mutations exhibit the same astrocyte defect? (2) How does impairment of EIF2B disrupt astrocyte biology? To address these questions, we will: (Aim 1) establish and characterize inducible pluripotent stem (iPS) cell lines from different VWM-disease patients, and (Aim 2) use these cells to examine astroglial differentiation, reactive gliosis and the ability to support survival and differentiation of oligodendrocytes.

消失性白色物质（VWM）病是最普遍的脑白质营养不良之一，并且由以下引起： 真核生物翻译起始因子EIF 2B五个亚基中任何一个的突变。尽管许多 尽管已经鉴定出不同类型的EIF 2B突变，但仍不清楚这些突变如何导致EIF 2B突变。 髓磷脂的大量流失 最大的问题之一是缺乏一个合适的实验系统来测试 EIF 2B突变对脑细胞的影响。我们对VWM患者来源的细胞的初步研究是通过以下方式实现的： 从VWM患者的大脑中分离活的胶质前体细胞的极其罕见的机会。本研究 发现了星形胶质细胞的缺陷但是缺乏合适的患者样本限制了我们研究细胞的能力。 导致并因此阻止我们找到治疗这种毁灭性疾病的方法。 该提案的目标是利用诱导性多能干细胞（iPSC）技术来获得神经干细胞。 来自具有不同EIF 2B复合突变的VWM患者的干细胞培养物。这些VWM神经干细胞将 用于检测星形胶质细胞分化和功能方面的基因型-表型关系。我们 初步数据表明，iPSC可以很容易地从VWM患者细胞中衍生出来，并且这些患者- 衍生的PSC可被诱导分化为神经干细胞和星形胶质细胞前体。突变 EIF 2B似乎引起细胞死亡增加，并损害一些星形胶质细胞谱系蛋白的表达， 包括星形胶质细胞谱系决定因子CREB 3L 1。 由于星形胶质细胞在维持中枢神经系统稳态中的关键作用，我们的发现提出了两个重要的 问题：（1）具有不同EIF 2B突变的VWM患者是否表现出相同的星形胶质细胞缺陷？(2)如何 EIF 2B损伤破坏星形胶质细胞生物学？为了解决这些问题，我们将：（目标1）建立和 表征来自不同VWM疾病患者的诱导型多能干（iPS）细胞系，和（目的2）使用 这些细胞检查星形胶质细胞分化，反应性胶质细胞增生和支持生存的能力， 少突胶质细胞的分化。