RNA Granules in Cerebellar Neurodegeneration

RNA 颗粒在小脑神经变性中的作用

基本信息

批准号：
9910465
负责人：
Daniel R Scoles
金额：
$ 45.49万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9910465
关键词：
Ablation Affect Aging Amyotrophic Lateral Sclerosis Ataxia Autophagocytosis Bacterial Artificial Chromosomes Binding Sites Brain Brain Stem Calcium Cause of Death Cells Cerebellum Cerebrum Cytoplasmic Granules Data Defect Dendrites Disease Disease model Drosophila genus Dyes Economic Burden Elements Fibroblasts Financial Hardship Fluorescent in Situ Hybridization Frequencies Functional disorder Genes Genetic Grant Half-Life Health Huntington Disease Image In Situ Hybridization In Vitro Inherited Investigation Label Malignant Neoplasms Measures Mediating Messenger RNA Metabolism Microscopy Modeling Motor Motor Neurons Movement Mus Mutation Myotonic dystrophy type 1 Nerve Degeneration Nervous System Heredodegenerative Disorders Neurodegenerative Disorders Neurons Oregon Pathogenesis Pathway interactions Patients Phenotype Physiology Population Production Protein Inhibition Proteins Proteome Proteomics Protocols documentation Publishing Purkinje Cells RNA RNA Binding RNA-Binding Proteins Research Resources Role SCA2 protein Slice Small Interfering RNA Spinocerebellar Ataxias Stress Structure Therapeutic Therapeutic Intervention Tissues Transgenic Mice Translations Type 2 Spinocerebellar Ataxia Work Yeasts base differential expression gain of function human disease improved in vivo in vivo monitoring induced pluripotent stem cell interdisciplinary approach lymphoblast mRNA Expression molecular phenotype mouse model mutant neurophysiology new therapeutic target overexpression polyglutamine protein TDP-43 stress granule targeted treatment therapeutic target trafficking transcriptome transcriptome sequencing

项目摘要

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 stress granules (SGs) in spinocerebellar ataxia type 2 (SCA2), a hereditary neurodegenerative disease affecting cerebellar Purkinje neurons (PNs) and other neurons in the cerebellum, brainstem and cerebrum. The cause of SCA2 is a gain-of-function CAG expansion in the ATXN2 gene resulting in an expanded polyglutamine (polyQ) in ataxin-2. ATXN2 was previously known to have functions in mRNA metabolism based on its interactions with multiple RNA binding proteins (RBPs) including A2BP1/RBFOX1, DDX6, PABP1, TDP-43, FUS. We have now demonstrated that mutant ATXN2 interacts with the SG protein Staufen1, and this interaction has specific effects on SGs. Staufen expression (but not DDX6 or PABPC1) is increased with ATXN2 mutation, and we have also identified specific downstream consequences of this interaction on the abundance of SG mRNAs directly interacting Staufen. The objective of the proposed research is to characterize mechanistically why Staufen expression increases with ATXN2 mutation, to characterize the proteomic and mRNA composition of SCA2 SGs in SCA2 patient fibroblasts and neurons and in SCA2 transgenic mice. SG elements that are identified by this work might be exploited therapeutically for treating SCA2. Three specific aims are proposed: 1) We will evaluate mutant ATXN2 inhibition of protein autophagy as a reason for staufen overexpression, and will characterize SCA2 SG rate of formation, half-life, and superstructure using superresolution microscopy, as well as SCA2 SG transcriptomes using purified SGs. 2) We will conduct FISH to localize SCA2 SG mRNAs in cultured SCA2 neurons and in cerebellar slices of SCA2 mice. 3) We will determine SCA2 mouse motor and neurophysiological phenotypes in SCA2 mice haploinsufficient for Staufen, and will record Purkinje cell firing frequencies simultaneously with imaging for TIA1 positive SGs and calcium abundance. This will determine whether abnormal SG trafficking/localization associates with abnormal PC physiology in SCA2 mice, depending on Staufen expression. The proposed work will clarify the role for SGs in neurodegeneration and will aid in the identification of new avenues towards treatments of SCA2 and other degenerative ataxias

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。这拟议的工作将表征 2 型脊髓小脑共济失调 (SCA2) 中的应激颗粒 (SG)，这是一种遗传性疾病影响小脑浦肯野神经元 (PN) 和小脑其他神经元的神经退行性疾病，脑干和大脑。 SCA2 的原因是 ATXN2 基因中的功能获得性 CAG 扩展导致 ataxin-2 中的聚谷氨酰胺 (polyQ) 扩展。 ATXN2 以前已知具有以下功能： mRNA 代谢基于其与多种 RNA 结合蛋白 (RBP) 的相互作用，包括 A2BP1/RBFOX1、DDX6、PABP1、TDP-43、FUS。我们现在已经证明突变体 ATXN2 与 SG 蛋白 Staufen1，这种相互作用对 SG 具有特定的影响。 Staufen 表达（但不是 DDX6 或 PABPC1) 随 ATXN2 突变而增加，我们还确定了特定的下游后果这种相互作用对直接与诗道芬相互作用的 SG mRNA 丰度的影响。拟议的目标研究的目的是从机制上描述为什么 Staufen 表达随着 ATXN2 突变而增加，表征 SCA2 患者成纤维细胞和神经元中 SCA2 SG 的蛋白质组和 mRNA 组成，在 SCA2 转基因小鼠中。这项工作确定的 SG 元素可能被用于治疗治疗SCA2。提出了三个具体目标：1）我们将评估突变体 ATXN2 对蛋白质的抑制作用自噬是诗道芬过度表达的一个原因，并将表征 SCA2 SG 的形成速率、半衰期、和超结构使用超分辨率显微镜，以及 SCA2 SG 转录组使用纯化的 SG。 2) 我们将进行 FISH 来定位培养的 SCA2 神经元和小脑切片中的 SCA2 SG mRNA。 SCA2 小鼠。 3) 我们将确定SCA2小鼠的运动和神经生理表型单倍体不足以满足诗道芬的要求，并且将在成像的同时记录浦肯野细胞放电频率 TIA1 阳性 SG 和钙丰度。这将确定 SG 贩运/本地化是否异常与 SCA2 小鼠中异常的 PC 生理学相关，具体取决于 Staufen 的表达。拟议的工作将阐明 SG 在神经退行性疾病中的作用，并有助于确定新的途径 SCA2 和其他退行性共济失调的治疗