SPOP modifies neurodegenerative proteinopathy in Alzheimer’s Disease.

SPOP 可以改善阿尔茨海默病中的神经退行性蛋白病。

基本信息

批准号：
10675938
负责人：
Randall Eck
金额：
$ 4.77万
依托单位：
UNIVERSITY OF WASHINGTON
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：
2023-09-16 至 2026-09-15
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10675938
关键词：
Adaptor Signaling Protein Age Age of Onset Aging Alleles Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease related dementia Amyloid beta-Protein Amyotrophic Lateral Sclerosis Automobile Driving Autopsy BRD2 gene Behavioral Binding Biological C9ORF72 CRISPR/Cas technology Caenorhabditis elegans Confocal Microscopy Cullin Proteins Data Dementia Deposition Development Diagnostic Dipeptides Disease Disease model Epigenetic Process Exhibits Frontotemporal Lobar Degenerations Functional disorder Future Genes Genetic Models Genetic Screening Homologous Gene Human Immunohistochemistry Link Liquid substance Longevity Mediating Messenger RNA Modeling Molecular Nerve Degeneration Neurodegenerative Disorders Neurofibrillary Tangles Neuronal Dysfunction Neurons Nuclear Nuclear RNA Pathologic Pathology Patients Phase Play Progressive Supranuclear Palsy Proteins Publishing RNA Splicing RNA interference screen Reporter Role Scaffolding Protein Shapes Signal Transduction System Tauopathies Testing Toxic effect Transgenic Animals Transgenic Model Ubiquitin Visualization Work cohort cullin-3 frontal lobe frontotemporal lobar dementia amyotrophic lateral sclerosis genetic approach hormonal signals hyperphosphorylated tau improved in vivo insight locomotor deficit loss of function loss of function mutation multicatalytic endopeptidase complex mutant neuron loss neurotoxicity novel novel therapeutics overexpression prevent protein TDP-43 protein degradation protein function sex tau Proteins tau aggregation tau-1 translational potential ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY Aggregated pathological tau protein constitutes one of the diagnostic hallmarks of Alzheimer’s disease (AD) and related disorders (ADRD). The molecular mechanisms by which pathological tau causes dysfunction and degeneration of neurons remain incompletely understood. However, pathological tau driven neuronal dysfunction and neurodegeneration clearly cause dementia. To investigate how pathological tau contributes to neurodegeneration in AD and ADRDs, we established a transgenic model in Caenorhabditis elegans for neurodegeneration driven by human tau aggregation. By employing classical forward genetic approaches, we identified several genes whose loss of function suppresses tauopathy, including spop-1. SPOP (speckle-type POZ protein) is a conserved nuclear adaptor protein for the RING E3 ubiquitin ligase Cullin-3 (CUL3), selecting proteins for degradation in the ubiquitin proteasome system (UPS). After generating a true null allele via CRISPR- cas9, we found that loss of spop-1 dramatically decreases tau accumulation and phosphorylation without altering tau mRNA abundance, rescues lifespan, reduces neurodegeneration, and improves behavioral deficits in tau transgenic animals. In addition to its role in UPS activity, SPOP also undergoes liquid-liquid phase separation (LLPS) localizing to nuclear speckles (NS). Our previously work in C. elegans and AD neurons indicate disruptions to NS function and composition contribute to tau toxicity. Remarkably, loss of spop-1 also rescues C9orf72 dipeptide-repeat toxicity in a C. elegans model of genetic amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Our preliminary results indicate loss of spop-1 suppresses behavioral deficits in C. elegans expressing ALS-linked mutant TDP-43 as well as transgenics co-expressing TDP-43 and tau, which recalculate AD and ADRD relevant interactions. Altogether, our work led us to hypothesize that SPOP functions as a multipurpose regulator of proteinopathy in AD and ADRDs. We hypothesize SPOP-CUL3 UPS activity and SPOP LLPS/NS localization contribute to the underlying mechanisms of tauopathy. To investigate these hypothesizes the specific aims of this project are: SPECIFIC AIMS: (1) Characterize SPOP loss of function in AD/ADRD neurodegenerative proteinopathies, (2) Determine the mechanism by which SPOP loss of function suppresses tauopathy, and (3) Evaluate the translational relevance of SPOP in Alzheimer’s disease. By completing the proposed work, we will uncover new molecular understandings of disease mechanisms including how SPOP protein – as well as nuclear speckles and Cullin-3 – participate in tauopathy and related ADRD co-pathologies. This work will also provide insight into the translational relevance of SPOP in Alzheimer’s disease brains, setting the stage for future study and informing the development of novel therapeutics.

项目概要聚集的病理性 tau 蛋白是阿尔茨海默病 (AD) 的诊断标志之一相关疾病 (ADRD) 病理性 tau 蛋白导致功能障碍的分子机制然而，神经元变性仍不完全清楚，病理性 tau 蛋白驱动的神经元功能障碍。和神经退行性变显然会导致痴呆。 AD 和 ADRD 中的神经退行性疾病，我们在秀丽隐杆线虫中建立了转基因模型由人类 tau 聚集驱动的神经退行性变通过采用经典的正向遗传方法，我们确定了一些功能丧失会抑制 tau 蛋白病的基因，包括 spop-1（斑点型）。 POZ 蛋白）是 RING E3 泛素连接酶 Cullin-3 (CUL3) 的保守核接头蛋白，选择通过 CRISPR 生成真正的无效等位基因后，在泛素蛋白酶体系统 (UPS) 中降解蛋白质。 cas9，我们发现 spop-1 的缺失会显着降低 tau 积累和磷酸化，而不会改变 tau mRNA 丰度，挽救寿命，减少神经退行性变，并改善 tau 行为缺陷转基因动物。除了在 UPS 活动中的作用外，SPOP 还经历液-液相分离 (LLPS)，定位于我们之前对秀丽隐杆线虫和 AD 神经元的研究表明 NS 功能受到破坏。和组成会导致 tau 毒性值得注意的是，spop-1 的丢失也挽救了 C9orf72 二肽重复序列。遗传性肌萎缩侧索硬化症 (ALS) 和额颞叶秀丽隐杆线虫模型的毒性我们的初步结果表明 spop-1 的缺失会抑制线虫的行为缺陷。表达 ALS 连锁突变体 TDP-43 以及共表达 TDP-43 和 tau 的转基因，重新计算 AD 和 ADRD 相关的相互作用。总而言之，我们的工作使我们重新认识到 SPOP 作为蛋白质病的多用途调节剂的功能我们勇敢地进行了 SPOP-CUL3 UPS 活动和 SPOP LLPS/NS 本地化贡献。为了研究这些进展，该项目的具体目标是：具体目标：(1) 描述 AD/ADRD 神经退行性蛋白病中 SPOP 功能丧失的特征， (2) 确定 SPOP 功能丧失抑制 tau 蛋白病变的机制，以及 (3) 评估 SPOP 在阿尔茨海默病中的转化相关性。通过完成拟议的工作，我们将揭示对疾病机制的新分子理解包括 SPOP 蛋白以及核斑点和 Cullin-3 如何参与 tau 蛋白病变及相关这项工作还将深入了解 SPOP 在阿尔茨海默病中的转化相关性。疾病大脑，为未来的研究奠定基础，并为治疗小说的发展提供信息。