Characterization of ATXN2 as a target for ALS in SCA2 motor neurons

ATXN2 作为 SCA2 运动神经元 ALS 靶标的表征

• 批准号: 9601486
• 负责人: Stefan M. PULST
• 金额: $ 19.06万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9601486
• 关键词: Affect; Aging; Algorithms; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Arsenites; Ataxia; Autophagocytosis; Biological Assay; Brain Stem; C9ORF72; CAG repeat; CRISPR/Cas technology; Cause of Death; Cell Line; Cells; Cellular Stress; Cerebellum; Clustered Regularly Interspaced Short Palindromic Repeats; Cytoplasm; Cytoplasmic Granules; Cytoplasmic Protein; DNA; Data; Dose; Economic Burden; Experimental Designs; Fibroblasts; Functional disorder; Funding; Future; Genes; Genetic; Grant; Health; Human; Individual; Inherited; Kainic Acid; Knockout Mice; Length; Link; Location; Longevity; Malignant Neoplasms; Measures; Messenger RNA; Metabolism; Modeling; Modification; Molecular; Motor; Motor Neurons; Mus; Mutate; Mutation; Nerve Degeneration; Nervous System Heredodegenerative Disorders; Neurodegenerative Disorders; Neurons; Nitric Oxide Donors; Pathologic; Pathology; Pathway interactions; Patients; Performance; Pharmaceutical Preparations; Phenotype; Physiology; Pluripotent Stem Cells; Population; Positioning Attribute; Proteins; Protocols documentation; Purkinje Cells; RNA; RNA-Binding Proteins; Research; Resources; Risk; Role; Skin; Spinal Cord; Stress; Testing; Thapsigargin; Therapeutic; Type 2 Spinocerebellar Ataxia; Work; base; burden of illness; dosage; experimental study; human disease; human pluripotent stem cell; improved; in vitro Model; induced pluripotent stem cell; inhibition of autophagy; molecular phenotype; mouse model; mutant; neuron loss; novel; polyglutamine; protein TDP-43; protein aggregate; response; superoxide dismutase 1; therapeutic target; therapy development; tool; transcriptome; transcriptomics

Project Summary/Abstract Neurodegenerative diseases represent an ever-increasing societal and economic burden with WHO estimates indicating that they will replace cancer as the 2nd leading cause of death by 2040. In neurodegenerative disease research, a wealth of pathways has been uncovered, but their direct and primary relevance to the respective human disease has been difficult to prove and targeting of pathways has remained difficult. The proposed work will characterize ATXN2 and pathways regulated by ATXN2 as therapeutic targets for amyotrophic lateral sclerosis (ALS), a disease burdening 1 out of 25,000 individuals. The ATXN2 gene is mutated in spinocerebellar ataxia type 2 (SCA2), a hereditary neurodegenerative disease affecting cerebellar Purkinje cells (PCs) and other neurons in the cerebellum and brainstem. Studies by us and others have demonstratedthat long normal CAG-repeat expansions in ATXN2 significantly increase the risk of ALS. More recently, survival (or lifespan) of some ALS mouse models and cultured ALS neuronal models were shown to be modified in association with ATXN2 mutation or overall level of expression. ATXN2 functions in mRNA metabolism based on its interactions with multiple RNA binding proteins (RBPs) including A2BP1/RBFOX1, DDX6, PABP1, and the ALS proteins TDP-43, FUS. Our most recent data demonstrate that mutant ATXN2 interacts with the stress granule (SG) RNA interacting protein Staufen1, and that this interaction has specific downstream effects on SGs. Since ALS motor neurons are also characterized by RNA granules, ATXN2 functions regulating staufen positive RNA granules in motor neurons may be relevant to ALS. The objective of this grant is to investigate ATXN2 as a therapeutic target for ALS using human pluripotent stem-cell-derived motor neurons from ALS and SCA2 patients. Three specific aims are proposed: 1) We will perform gene editing a panel of pluripotent stem cell lines (“CRISPR editing”) creating multiple pairs of cells that are genetically identical (“isogenic”) except for the edited change at of the ATXN2 gene. 2) We will then extensively characterize the phenotype of these cells vs ALS cells when differentiated to motor neurons neurons. 3) We will determine how lowering the expression of ATXN2 using our existing ATXN2 antisense oligonucleotide DNA drug alters the phenotypes of patient motor neurons generated in aim 1, including the dynamics of RNA granules in the presence of ATXN2 mutation. The proposed work will clarify the role for SGs in neurodegeneration and will aid in the identification of new avenues toward treatments of ALS and other degenerative ataxias including SCA2.

项目总结/摘要 据世卫组织估计，神经退行性疾病是一种日益加重的社会和经济负担 到2040年，它们将取代癌症成为第二大死亡原因。在神经退行性 疾病的研究，丰富的途径已经被发现，但他们的直接和主要的相关性， 各种人类疾病难以证实，并且靶向途径仍然很困难。的 拟议的工作将描述ATXN 2和ATXN 2调节的途径作为治疗靶点， 肌萎缩性侧索硬化症（ALS）是一种每25，000人中就有1人患有的疾病。ATXN 2基因是 脊髓小脑性共济失调2型（SCA 2）突变，SCA 2是一种影响小脑的遗传性神经退行性疾病， 小脑和脑干中的浦肯野细胞（PC）和其他神经元。我们和其他人的研究 证明ATXN 2中长的正常CAG重复扩增显著增加ALS的风险。更 最近，一些ALS小鼠模型和培养的ALS神经元模型的存活（或寿命）显示， 与ATXN 2突变或总体表达水平相关的修饰。ATXN 2在mRNA中的功能 基于其与多种RNA结合蛋白（RBP）（包括A2 BP 1/RBFOX 1）的相互作用的代谢， DDX 6、PABP 1和ALS蛋白TDP-43、FUS。我们最新的数据表明，突变ATXN 2 与应激颗粒（SG）RNA相互作用蛋白Staufen 1相互作用，并且这种相互作用具有特异性 对SG的下游影响。由于ALS运动神经元也以RNA颗粒为特征，ATXN 2 调节运动神经元中施陶芬阳性RNA颗粒的功能可能与ALS有关。的目标 该基金将研究ATXN 2作为ALS的治疗靶点， ALS和SCA 2患者的运动神经元。提出了三个具体目标：1）我们将执行基因 编辑一组多能干细胞系（“CRISPR编辑”），产生多对细胞， 除了ATXN 2基因的编辑改变之外，它们在遗传上是相同的（“同基因的”）。2)我们将广泛 表征这些细胞相对于ALS细胞在分化为运动神经元神经元时的表型。3)我们 将确定如何使用我们现有的ATXN 2反义寡核苷酸DNA降低ATXN 2的表达 药物改变了目标1中产生的患者运动神经元的表型，包括RNA动力学 在存在ATXN 2突变的情况下的颗粒。拟议的工作将澄清秘书长在以下方面的作用： 神经退行性变，并将有助于确定治疗ALS和其他疾病的新途径 退行性共济失调包括SCA 2。