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.

蛋白质和RNA通过核孔的核细胞质转运（NCT）最近出现了作为神经变性的中心机制。确实，我们最近已经表明，突变的亨廷顿明显加剧核完整性和NCT破坏的衰老相关改变，以及核孔介导的缺陷在C9ORF72相关的肌萎缩性侧索硬化症（ALS）和额颞痴呆症中也发现了运输（FTD）。在这里，我们证明了突变FUS的表达，一种与ALS和FTD相关的蛋白质的表达导致核丧失孔完整性，改变核包膜形态和与多个相关的NCT中断有关的其他表型包括等源性人神经元和新型FUS敲入小鼠模型的模型。在这个项目中，我们将进一步使用ALS和FTD中与疾病相关的FUS和NCT受损受损之间的关系上述模型系统和人类CNS组织（AIM 1）。阐明表达的机制突变FUS诱导与NCT相关的表型，我们将进一步探测FUS与核孔之间的相互作用蛋白质，并检验以下假设：异常的相变涉及突变FUS和核孔蛋白有助于破坏的NCT（AIM 2）。我们还认为，NCT损害通过错误定位的毒性诱导毒性笔录和蛋白质，这是通过使用RNA测序和蛋白质组学来比较基因表达的概念在NCT相关的表现之前和之后，突变FUS与对照神经元中的细胞分级分馏后表型（目标3）。最后，我们将测试靶向人类神经元中核出口的治疗潜力，使用已经证明在多种人类疾病中既安全又有效的策略的小鼠（AIM 4）。两个学术团体与工业合作伙伴之间的这种协同合作（谁将提供治疗在我们的实验室研究的化合物）有可能发现新的机械洞察并建立靶向ALS/FTD中核细胞质转运的治疗潜力。