NUP50 as a modifier and risk factor for TDP-43 pathology in FTD/ALS

NUP50 作为 FTD/ALS 中 TDP-43 病理的修饰剂和危险因素

基本信息

批准号：
10800366
负责人：
Wilfried Rossoll
金额：
$ 43.03万
依托单位：
MAYO CLINIC JACKSONVILLE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-30 至 2025-09-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10800366
关键词：
ALS pathology ALS patients Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Amyotrophic Lateral Sclerosis Animal Model Autopsy Brain Cell Nucleus Cell model Cell physiology Cells Cytoplasm Data Data Set Defect Detergents Disease Disease model Frontotemporal Dementia Functional disorder Future Genes Human Immunohistochemistry Impairment In Vitro Induced pluripotent stem cell derived neurons Injections Intracellular Transport Knowledge Link Mass Spectrum Analysis Modeling Molecular Molecular Chaperones Morphology Mutation Nerve Degeneration Neurodegenerative Disorders Nuclear Nuclear Pore Nuclear Pore Complex Nuclear Pore Complex Proteins Outcome Pathogenesis Pathologic Pathology Pathway interactions Patients Phase Phase Transition Physiological Process Protein Import Proteins Proteomics RNA RNA Processing RNA-Binding Proteins Research Research Support Ribonucleoproteins Risk Factors Role Slice Solubility TDP-43 aggregation Testing Therapeutic Toxic effect Variant age related neurodegeneration brain tissue design effective therapy experimental study frontotemporal lobar dementia amyotrophic lateral sclerosis human disease in vitro activity in vivo in vivo Model insight lymphoblastoid cell line mouse model mutant neuronal survival neuropathology new therapeutic target novel nucleocytoplasmic transport pre-clinical prion-like protein TDP-43 protein aggregation protein function proteotoxicity stress granule targeted treatment therapeutic target therapy development

项目摘要

The cytoplasmic mislocalization and accumulation of insoluble and misfolded RNA- binding protein TDP-43 is the neuropathological hallmark of the amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) disease spectrum but is also commonly found in Alzheimer’s disease (AD) and other AD-related dementias (ADRDs), marking TDP-43 mislocalization as a key pathomechanism and high priority target for therapy development. Previous studies from our lab and others have established nuclear pore defects and impaired protein import and RNA export, as a common hallmark of FTD/ALS and other age-related neurodegenerative diseases. While there is growing evidence that perturbations in the nucleocytoplasmic transport machinery are linked to TDP-43 pathology, the causal relationship between intracellular transport pathways and disease pathogenesis is not well understood. As prior research to support the premise of this proposal, we have discovered that expression of the nucleoporin NUP50 in FTD/ALS models can restore solubility and nuclear localization of TDP-43, suggesting an important role for NUP50 in the disease process. This premise is further supported by the recent discovery of rare NUP50 variants as a risk factor for ALS, and a critical role for NUP50 in neuronal survival. These findings lead us to hypothesize that NUP50 serves in a non-canonical role in reducing protein mislocalization and aggregation, while its deficiency in FTD/ALS may lead to increased TDP-43 pathology, suggesting NUP50 as a promising target for therapy development. To test this hypothesis, we will use TDP-43 cellular, organotypic and animal models, patient-derived and induced pluripotent stem cell (iPSC)-derived neurons, in combination with mass spectrometry (MS)-based quantitative proteomics and immunohistochemistry in human brain tissue, to elucidate how NUP50 can reduce TDP-43 pathology, and how ALS/FTD-associated mutations in NUP50 contribute towards TDP-43 pathology. In Aim 1 we will methodically investigate the effect of NUP50 on aberrant TDP-43 phase transition and mislocalization and establish the role of structural domains and associated proteins for a mechanistic understanding of this activity in vitro. In Aim 2 we will determine the effect of NUP50 on TDP-43-dependent neurodegeneration in FTD/ALS mouse models, to evaluate NUP50 and associated proteins as a potential therapeutic targe in vivo. In Aim 3, we will establish the role of NUP50 in human TDP-43 proteinopathies, by testing the effect of NUP50 deficiency and disease-associated NUP50 variants on TDP-43 pathology and assessing the abundance and localization of NUP50 in FTD/ALS and ADRD autopsy brain tissue. Our findings will impact the field by providing a detailed mechanistic understanding of how NUP50 expression can impact TDP-43 pathology in disease models, and by establishing the role of NUP50 in the disease process and validating it as a potential therapeutic target in FTD/ALS and other ADRDs.

不溶性和错误折叠的RNA-的细胞质错误定位和积累结合蛋白TDP-43是肌萎缩性侧面硬化症（ALS）和额颞痴呆（FTD）疾病谱，但在阿尔茨海默氏病（AD）和其他与广告相关的痴呆症（ADRDS），将TDP-43错误定位标记为关键病理机理和较高治疗开发的优先目标。我们实验室和其他人的先前研究已经建立了核孔缺陷和蛋白质进口的障碍 RNA导出，是FTD/ALS和其他与年龄有关的神经退行性疾病的共同标志。尽管越来越多的证据表明，核眼质运输机制中的扰动与TDP-43有关病理学，细胞内转运途径与疾病发病机理之间的因果关系不好理解。作为支持该提议前提的先前研究，我们发现了这种表达 FTD/ALS模型中的核动脉蛋白NUP50可以恢复TDP-43的可溶性和核定位，提出NUP50在疾病过程中的重要作用。最近的前提得到了进一步的支持发现稀有的NUP50变体是ALS的危险因素，而NUP50在神经元存活中的关键作用。这些发现使我们假设NUP50在降低蛋白质中起着非规范作用错误的定位和聚集，而其在FTD/ALS中的缺乏可能会导致TDP-43病理增加，但建议NUP50作为治疗开发的承诺目标。为了检验该假设，我们将使用TDP-43细胞，有机和动物模型，患者衍生和诱导多能干细胞（IPSC）衍生的神经元，结合质谱（MS）的神经元定量蛋白质组学和人脑组织中的免疫组织化学，以阐明NUP50如何减少 TDP-43病理学以及NUP50中ALS/FTD相关的突变如何有助于TDP-43病理学。在AIM 1中，我们将有条不紊地研究NUP50对异常TDP-43相变的影响错误定位并确定结构域和相关蛋白的作用在体外了解这种活动。在AIM 2中，我们将确定NUP50对TDP-43依赖性的影响 FTD/ALS小鼠模型中的神经变性，以评估NUP50和相关蛋白作为潜力体内治疗靶标。在AIM 3中，我们将通过测试NUP50缺乏症和与疾病相关的NUP50变体对TDP-43病理和评估NUP50在FTD/ALS和ADRD尸检脑组织中的抽象和定位。我们的发现将通过对NUP50的表达方式提供详细的机械理解来影响该领域可以影响疾病模型中的TDP-43病理，并通过确定NUP50在疾病过程中的作用并将其验证为FTD/ALS和其他ADRD的潜在治疗靶标。