Regulation of paraspeckles by STAU1 in neurodegenerative disease

STAU1 在神经退行性疾病中对 paraspeckles 的调节

• 批准号: 10668027
• 负责人: Daniel R Scoles
• 金额: $ 19.23万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10668027
• 关键词: ALS patients; Affect; Aging; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Apoptosis; Attenuated; Autophagocytosis; Bacterial Artificial Chromosomes; Binding; C9ORF72; CASP3 gene; Cell Culture Techniques; Cell Line; Cell Nucleus; Cell model; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Core Protein; Cortical Cord; Cytoplasm; Data; Disease; Disease model; Evaluation; Fibroblasts; Financial Hardship; Frontotemporal Dementia; Genes; Health; Human; Hyperactivity; In Vitro; Knockout Mice; Laboratories; Liquid substance; Mass Spectrum Analysis; Mediating; Messenger RNA; MicroRNAs; Molecular; Motor; Mus; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear; Nuclear Translocation; Pathology; Pathway interactions; Patients; Phase; Phenotype; Population; Proteins; RNA-Binding Proteins; Regulation; Reporter; Role; Safety; Small Interfering RNA; Spinal Cord; Stress; Therapeutic; Translating; Translational Repression; Translations; Type 2 Spinocerebellar Ataxia; Untranslated RNA; Western Blotting; differential expression; frontotemporal lobar dementia amyotrophic lateral sclerosis; heart function; improved; in vivo Model; induced pluripotent stem cell; interest; mRNA Transcript Degradation; mRNA Translation; motor behavior; mouse model; mutant; overexpression; protein TDP-43; sporadic amyotrophic lateral sclerosis; therapeutic target; trafficking; transcriptome

Project Summary/Abstract Nuclear paraspeckles function to regulate mRNA translation. mRNAs that are sequestered in paraspeckles are translationally silent while those not entering paraspeckles can translocate from the nucleus where the can effectively be translated. There is now accumulating evidence that paraspeckles are increased in number and size in neurodegenerative diseases, including Alzheimer’s disease (AD), frontotemporal dementia (FTD), and amyotrophic lateral sclerosis (ALS). RNA-binding proteins (RBPs) have a central role in neurodegenerative disease, but their functions in paraspeckles are not well understood, yet we confirmed a critical regulatory role of paraspeckles mediated by the stress-related RBP STAU1. In supporting studies, we discovered that STAU1 is overabundant in multiple neurodegenerative diseases including AD, FTD, and ALS, and that by normalizing STAU1 various molecular and motor phenotypes relevant to these diseases are improved or restored. Relevant to paraspeckles, STAU1 competes for mRNA binding at inverted repeat Alu sequences (IRAlus) with the paraspeckle core protein p54nrb and directs mRNA translocation to the cytoplasm, for active translation. The long noncoding RNA (lncRNA) NEAT1 is indispensable for paraspeckle formation, involving p54nrb interaction. In our preliminary studies we found that NEAT1 is strikingly downregulated in cells depleted of STAU1. NEAT1 also interacts with TDP-43 and is overabundant in AD, FTD, and ALS. Since both NEAT1 and STAU1 are overabundant in multiple neurodegenerative diseases, we hypothesize that normalization of STAU1 abundance will restore NEAT1 to normal levels, thereby restoring paraspeckle function. STAU1 will likely be a better therapeutic target than NEAT1 itself, because mice null for Neat1 have reduced cardiac function while mice null for Stau1 are viable and are characterized by no neurodegeneration. Our proposal is divided into two specific aims. The first aim will determine NEAT1 levels and paraspeckle localization as well as paraspeckle sizes and numbers in cell models that have been edited to express mutant ATXN2, that interacts with STAU1 in cytoplasmic phase-separated liquid droplets characterized by overabundant STAU1. We also evaluate Neat1 abundance and localization in a mouse model that we developed that overexpresses STAU1. In the second aim we characterize paraspeckles and NEAT1 in cultured human cortical neurons, with focus on TDP-43 pathology. The successful demonstration that NEAT1 is altered in diseases characterized by STAU1 overabundance will lead to new potential therapeutic targets for AD, FTD, ALS and other disorders with abnormal nuclear paraspeckle function, deeper understanding on the significance of NEAT1 and STAU1 to neurodegenerative disease, and will further support STAU1 as a therapeutic target.

项目概要/摘要 核 paraspeckles 的功能是调节 mRNA 翻译，这些 mRNA 被隔离在 paraspeckles 中。 平移沉默，而那些未进入paraspeckles的细胞可以从细胞核转位，在那里 现在有越来越多的证据表明副斑雀的数量和数量有所增加 神经退行性疾病的大小，包括阿尔茨海默病 (AD)、额颞叶痴呆 (FTD) 和 肌萎缩性脊髓侧索硬化症 (ALS) 在神经退行性疾病中发挥着重要作用。 疾病，但它们在副斑点中的功能尚不清楚，但我们证实了关键的调节作用 在支持研究中，我们发现 STAU1 介导的副斑点。 在包括 AD、FTD 和 ALS 在内的多种神经退行性疾病中过量存在，并且通过正常化 STAU1 与这些疾病相关的各种分子和运动表型得到改善或恢复相关。 对于 paraspeckles，STAU1 与反向重复 Alu 序​​列 (IRAlus) 竞争 mRNA 结合 paraspeckle 核心蛋白 p54nrb 并指导 mRNA 易位至细胞质，进行主动翻译。 长非编码 RNA (lncRNA) NEAT1 对于副斑形成不可或缺，涉及 p54nrb 相互作用。 我们的初步研究发现 NEAT1 在 NEAT1 耗尽的细胞中显着下调。 也与 TDP-43 相互作用，并且在 AD、FTD 和 ALS 中过量，因为 NEAT1 和 STAU1 都是。 STAU1 丰度在多种神经退行性疾病中过多，我们面临着 STAU1 丰度正常化的问题 将使 NEAT1 恢复到正常水平，从而恢复 STAU1 功能可能会更好。 治疗靶点高于 NEAT1 本身，因为 Neat1 缺失的小鼠心脏功能下降，而 NEAT1 缺失的小鼠心脏功能下降 对于Stau1来说是可行的并且特点是没有神经变性我们的建议分为两个具体的。 第一个目标将确定 NEAT1 水平和副散斑定位以及副散斑大小和 细胞模型中的数字已被编辑以表达突变体 ATXN2，该突变体与 STAU1 相互作用 我们还评估了 Neat1 的细胞质相分离液滴。 我们开发的过度表达 STAU1 的小鼠模型中的丰度和定位。 我们对培养的人类皮质神经元中的 paraspeckles 和 NEAT1 进行了表征，重点关注 TDP-43 病理学。 成功证明 NEAT1 在以 STAU1 过多为特征的疾病中发生改变将 为 AD、FTD、ALS 和其他核异常疾病带来新的潜在治疗靶点 paraspeckle功能，更深入地了解NEAT1和STAU1对神经退行性疾病的意义 疾病，并将进一步支持 STAU1 作为治疗靶点。