权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Disruption of nucleocytoplasmic transport in FUS-related neurodegenerative diseases

FUS 相关神经退行性疾病中核细胞质运输的破坏

基本信息

批准号：
9927700
负责人：
Daryl Angela Bosco
金额：
$ 81.49万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-15 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927700
关键词：
Affect Age Aging Amyotrophic Lateral Sclerosis Autopsy Biological Models Brain C9ORF72 Calpain Cell Culture Techniques Cell Fractionation Cell Line Cell Nucleus Cellular Stress Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complex Cytoplasm DNA Damage DNA-Binding Proteins Data Defect Disease Exhibits Fractionation Gene Expression Genomics Goals Human Huntington gene Impairment In Vitro Industrialization Inherited Knock-in Knock-in Mouse Laboratories Liquid substance Literature Mediating Modeling Morphology Motor Neurons Mus Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Nuclear Nuclear Envelope Nuclear Export Nuclear Pore Nuclear Pore Complex Nuclear Pore Complex Proteins Nucleocytoplasmic Transport Proteins Pathogenicity Pathway interactions Patients Phase Phase Transition Phenotype Pore Proteins Process Property Proteins Proteome Proteomics Publishing RNA RNA Transport Recombinant Proteins Research Stress Testing Therapeutic Tissues Toxic effect Transcript age related calpain inhibitor cell age density exportin 1 protein frontotemporal lobar dementia-amyotrophic lateral sclerosis human disease induced pluripotent stem cell industry partner inhibitor/antagonist insight link protein mouse model mutant novel nucleocytoplasmic transport therapeutic evaluation therapeutic target transcriptome sequencing

项目摘要

Impaired nucleocytoplasmic transport (NCT) of protein and RNA through the nuclear pore has recently emerged as a central mechanism in neurodegeneration. Indeed, we have recently shown that mutant huntingtin markedly exacerbates aging-related alterations in nuclear integrity and disruption of NCT, and defects in nuclear pore-mediated transport have also been uncovered in C9ORF72-related amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here, we demonstrate that expression of mutant FUS, a protein linked to ALS and FTD, leads to a loss of nuclear pore integrity, altered nuclear envelope morphology and other phenotypes related to disrupted NCT in multiple, relevant models including isogenic human neurons and a novel FUS knock-in mouse model. In this project, we will further interrogate the relationship between disease-associated FUS and impaired NCT in ALS and FTD using the aforementioned model systems and human CNS tissues (Aim 1). To elucidate the mechanism by which expression of mutant FUS induces NCT-related phenotypes, we will further probe the interactions between FUS and nuclear pore proteins, and test the hypothesis that abnormal phase transitions involving mutant FUS and nuclear pore proteins contribute to disrupted NCT (Aim 2). We also posit that impaired NCT induces toxicity via the mislocalization of transcripts and proteins, a notion that will be tested by using RNA-sequencing and proteomics to compare gene expression after cellular fractionation in mutant FUS versus control neurons, before and after manifestation of NCT-related phenotypes (Aim 3). Finally, we will test the therapeutic potential of targeting nuclear export in FUS human neurons and mice using strategies that have already been shown to be both safe and effective across multiple human diseases (Aim 4). This synergistic collaboration between two academic groups and an industrial partner (who will provide the therapeutic compound for studies in our laboratories) has the potential to uncover new mechanistic insights to disease and establish the therapeutic potential of targeting nucleocytoplasmic transport in ALS/FTD.

最近出现了蛋白质和 RNA 通过核孔的核细胞质转运 (NCT) 受损的情况作为神经退行性变的核心机制。事实上，我们最近表明突变亨廷顿蛋白显着加剧与衰老相关的核完整性改变和 NCT 破坏，以及核孔介导的缺陷在 C9ORF72 相关的肌萎缩侧索硬化症 (ALS) 和额颞叶痴呆中也发现了转运（FTD）。在这里，我们证明突变 FUS（一种与 ALS 和 FTD 相关的蛋白质）的表达会导致核孔完整性、核膜形态改变以及与多个相关 NCT 破坏相关的其他表型模型包括同基因人类神经元和新型 FUS 敲入小鼠模型。在这个项目中，我们将进一步使用以下方法询问 ALS 和 FTD 中疾病相关的 FUS 与 NCT 受损之间的关系上述模型系统和人类中枢神经系统组织（目标 1）。阐明表达的机制突变FUS诱导NCT相关表型，我们将进一步探讨FUS与核孔之间的相互作用蛋白质，并检验涉及突变 FUS 和核孔蛋白的异常相变的假设为破坏 NCT 做出贡献（目标 2）。我们还假设受损的 NCT 通过错误定位来诱导毒性转录本和蛋白质，这一概念将通过使用 RNA 测序和蛋白质组学来比较基因表达进行测试突变 FUS 与对照神经元的细胞分级后，NCT 相关表现之前和之后表型（目标 3）。最后，我们将测试 FUS 人类神经元中靶向核输出的治疗潜力小鼠使用已被证明对多种人类疾病既安全又有效的策略（目标 4）。两个学术团体和一个工业合作伙伴（将提供治疗药物）之间的协同合作用于我们实验室研究的化合物）有可能发现疾病的新机制见解并建立靶向核细胞质转运在 ALS/FTD 中的治疗潜力。