Contribution of astrocytes to mutant FUS-linked Amyotrophic Lateral Sclerosis

星形胶质细胞对突变型 FUS 相关肌萎缩侧索硬化症的影响

基本信息

批准号：
10160975
负责人：
PIERA PASINELLI
金额：
$ 34.13万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10160975
关键词：
AMPA Receptors Adult Affect Amyotrophic Lateral Sclerosis Astrocytes Autocrine Communication Automobile Driving Autopsy Biological Assay Biology Brain Cause of Death Cell Death Cell Survival Cells Cessation of life Complex Conditioned Culture Media DNA Development Disease Disease Progression Environment Exposure to Feedback Functional disorder Genes Genotype Goals Human In Vitro Inflammatory Link Measures Mediating Messenger RNA Modeling Molecular Motor Motor Neurons Mus Mutation Neurodegenerative Disorders Neuroglia Neurons Nuclear Pathogenesis Pathogenicity Pathologic Pathology Pathway interactions Patients Pharmacology Phenotype Physiology Play Proteins Publishing RNA-Binding Proteins Reporter Rodent Role Signal Transduction Spatial Distribution Spinal Cord Standardization System TNF gene TNFRSF1A gene Testing Therapeutic Time Tissues Toxic effect base cell injury comparative cytokine excitotoxicity experimental study fused in sarcoma genetic manipulation human disease in vivo in vivo Model individualized medicine induced pluripotent stem cell motor behavior motor disorder motor impairment mouse model mutant mutant mouse model neuron loss neurotoxic overexpression prevent promoter single cell analysis sporadic amyotrophic lateral sclerosis vector

项目摘要

ABSTRACT The goal of this project is to identify the mechanisms by which mutations in Fused in Sarcoma (FUS) induce pathological and pathogenic changes in astrocytes in FUS-linked Amyotrophic Lateral Sclerosis (FUS-ALS). Astrocytes are known to contribute to disease progression in some, but not all, forms of ALS and, until recently, not much was known about the role of astrocytes in FUS-ALS. Our lab has recently published the first in vitro evidence that astrocytes expressing mutant forms of FUS (mutFUS), but not wild-type FUS, exert toxicity on motor neurons through activation of the NFkB pathway leading to the release of secreted factor(s), and primarily TNFα (Kia & McAvoy et al., GLIA, 2018). In a TNFα−dependent manner, motor neurons exposed to the conditioned medium of mutFUS astrocytes show AMPA receptor alterations that sensitize them to excitotoxic damage, leading to cell death. Activation of astrocytic NFkB and TNFα release seems to be specific to mutFUS-ALS, underscoring the importance of dissecting disease mechanisms specific to each form of ALS in order to develop tailored therapies. In this proposal, we will focus on mutFUS-ALS and will study (1) the mechanisms by which disease-casuative mutations in FUS alter astrocyte biology and (2) how mutant FUS expressing astrocyte affect the viability of other cells in the spinal cord in vivo. The ultimate goal is to identify key pathways targeted by mutFUS to eventually develop specific therapies. Ultimately, our models of FUS-ALS may serve as a platform for a comparative analysis with models mimicking other forms of ALS, so to identify genotype- specific vs. converging and more global mechanisms of disease pathogenesis. For our studies, we will use in vitro and in vivo models consisting of (1) primary rodent astrocytes transduced with different FUS mutations as well as iPSc-derived astrocytes from FUS patients; (2) an in vivo mouse model of mutFUS ALS. The in vitro systems will allow us to dissect the precise mechanisms by which mutFUS targets the astrocytes and to determine how mutFUS alters astrocyte biology in patients. The mouse model will allow us to study how mutFUS astrocytes affect the cellular environment in a complex in vivo setting.

抽象的该项目的目的是确定融合中突变的机制肉瘤（FUS）诱导FUS连接的星形胶质细胞的病理和致病性变化肌萎缩性侧索硬化症（FUS-ALS）。已知星形胶质细胞在某些但不是全部的形式中有助于疾病进展 ALS，直到最近，星形胶质细胞在FUS-ALS中的作用还不多。我们的实验室最近发布了表达星形胶质细胞的第一个体外证据 FUS的突变形式（mutfus），而不是野生型FUS，对运动神经元发挥毒性通过激活NFKB途径，导致分泌因子的释放和以TNFα依赖的方式，电动机暴露于Mutfus星形胶质细胞条件介质的神经元显示AMPA接收器将它们感知到兴奋性毒性损伤，导致细胞死亡。激活星形胶质细胞NFKB和TNFα释放似乎特定于Mutfus-Als，强调剖析针对每种形式的ALS的疾病机制的重要性开发量身定制的疗法。在此提案中，我们将重点关注mutfus-als，并将研究（1） FUS中哪些疾病生产性突变改变了星形胶质细胞生物学，（2）突变体如何表达星形胶质细胞的FU会影响体内脊髓中其他细胞的活力。这最终目标是确定Mutfus瞄准的关键途径以最终发展具体疗法。最终，我们的Fus-Als模型可以作为一个平台与模仿其他形式的ALS的模型进行比较分析，因此要鉴定基因型 - 疾病发病机理的特定与融合和更多全球机制。为我们研究，我们将使用由（1）原发啮齿动物星形胶质细胞组成的体外和体内模型用不同的FUS突变以及IPSC衍生的星形胶质细胞翻译患者; （2）mutfus als的体内小鼠模型。体外系统将使我们剖析Mutfus靶向星形胶质细胞的精确机制和确定Mutfus如何改变患者的星形胶质细胞生物学。鼠标模型将允许美国研究Mutfus星形胶质细胞如何影响体内复杂的体内细胞环境环境。