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.

项目摘要/摘要 核paraspeccckles功能调节mRNA翻译。在paraspecckles中隔离的mRNA是 在不输入paraspecckles的那些人可以从细胞核中翻译而来的静音 有效地翻译。现在有累积的证据表明paraspeccckles数量增加，并且 神经退行性疾病的大小，包括阿尔茨海默氏病（AD），额颞痴呆（FTD）和 肌萎缩性侧索硬化症（ALS）。 RNA结合蛋白（RBP）在神经退行性中具有核心作用 疾病，但是它们在paraspecckles中的功能尚不清楚，但我们确认了关键的调节作用 由应力相关的RBP STAU1介导的paraspeccckles。在支持研究中，我们发现Stau1 在包括AD，FTD和ALS在内的多种神经退行性疾病中过多，并且通过归一化 与这些疾病相关的各种分子和运动表型得到了改善或恢复。相关的 对于paraspeccckles，stau1竞争在倒重复序列（iralus）与 paraspecckle核心蛋白p54nrb并将mRNA转移到细胞质中，以进行主动翻译。这 长的非编码RNA（LNCRNA）Neat1对于paraspecckle的形成是必不可少的，涉及p54NRB相互作用。在 我们的初步研究发现，Neat1在耗尽stau1的细胞中明显下调。 Neat1 还与TDP-43相互作用，并且在AD，FTD和ALS中过多。由于Neat1和Stau1都是 在多种神经退行性疾病中过多，我们假设stau1抽象的归一化 将将Neat1恢复到正常水平，从而恢复paraspecckle函数。 Stau1可能会更好 治疗靶标比Neat1本身，因为NEAT1的小鼠无效心脏功能降低，而小鼠无效 因为stau1是可行的，并以无神经变性为特征。我们的建议分为两个特定的 目标。第一个目标将确定Neat1级别和paraspecckle定位以及paraspecckle的大小和 已编辑以表达突变atxn2的细胞模型中的数字，与stau1相互作用 细胞质分离的液滴，其特征是过多的stau1。我们还评估Neat1 我们开发过过表达stau1的小鼠模型中的抽象和定位。在第二个目标 我们在培养的人皮质神经元中表征了paraspeccckles和neat1，重点是TDP-43病理学。 成功的证明Neat1在以STAU1过度为特征的疾病中改变了 导致针对AD，FTD，ALS和其他核异常疾病的新潜在治疗靶标 拼贴功能，更深入地了解Neat1和stau1对神经退行性的重要性 疾病，并将进一步支持STAU1作为治疗靶点。