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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要根据世界卫生组织的估计，神经退行性疾病代表着不断增加的社会和经济负担表明到 2040 年它们将取代癌症成为第二大死因。疾病研究中，已经发现了大量的途径，但它们与疾病有直接和主要的相关性。各自的人类疾病很难被证明，并且靶向途径仍然很困难。拟议的工作将描述 ATXN2 和 ATXN2 调节的通路作为治疗靶点肌萎缩侧索硬化症 (ALS) 是一种疾病，每 25,000 人中就有 1 人患有 ATXN2 基因。脊髓小脑共济失调 2 型 (SCA2) 发生突变，这是一种影响小脑的遗传性神经退行性疾病我们和其他人的研究表明，小脑和脑干中的浦肯野细胞（PC）和其他神经元。 ATXN2 中显示的长时间正常 CAG 重复扩增会显着增加 ALS 的风险。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）被证明与 ATXN2 突变或 ATXN2 mRNA 功能的总体表达水平相关的修饰。基于其与多种 RNA 结合蛋白 (RBP) 相互作用的代谢，包括 A2BP1/RBFOX1， DDX6、PABP1 和 ALS 蛋白 TDP-43、FUS 我们最新的数据表明突变 ATXN2。与应激颗粒 (SG) RNA 相互作用蛋白 Staufen1 相互作用，并且这种相互作用具有特异性由于 ALS 运动神经元也以 RNA 颗粒为特征，ATXN2 具有下游作用。运动神经元中staufen阳性RNA颗粒的调节功能可能与ALS相关。这笔赠款将利用人类多能干细胞来研究 ATXN2 作为 ALS 的治疗靶点提出了三个具体目标：1）我们将进行基因研究。编辑一组多能干细胞系（“CRISPR 编辑”），创建多对细胞，这些细胞基因相同（“同基因”），除了 ATXN2 基因的编辑变化 2) 我们通常会。表征这些细胞与 ALS 细胞分化为运动神经元时的表型 3) 我们。将确定如何使用我们现有的 ATXN2 反义寡核苷酸 DNA 降低 ATXN2 的表达药物改变目标 1 中产生的患者运动神经元的表型，包括 RNA 的动态存在 ATXN2 突变的颗粒拟议的工作将阐明 SGs 在 ATXN2 突变中的作用。神经退行性疾病，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 SCA2。