RNA Surveillance and Protein Translation in FTD

FTD 中的 RNA 监测和蛋白质翻译

基本信息

批准号：
10687846
负责人：
Haining Zhu
金额：
$ 53.11万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10687846
关键词：
Acetylation Affect Age Amyotrophic Lateral Sclerosis Animal Model Cell Culture Techniques Cell Nucleus Cell model Cell physiology Clinical Cultured Cells Cytoplasm Disease Disease Progression Etiology Frontotemporal Dementia Frontotemporal Lobar Degenerations Functional disorder Future Genetic Impairment In Vitro Label Laboratory Finding Light Liquid substance Lobe Measures Mediating Membrane Messenger RNA Methods Molecular Mus Mutation Names Nerve Degeneration Neuronal Dysfunction Neurons Nonsense-Mediated Decay Pathogenesis Pathogenicity Pathologic Pathology Pathway Analysis Pathway interactions Patients Phase Play Polyribosomes Post-Translational Protein Processing Process Prosencephalon Proteins Proteomics Protocols documentation Publishing Puromycin RNA RNA Binding Research Ribonucleoproteins Role System Temporal Lobe Testing Tissues Toxic effect Transgenic Mice Translating Translations Work age related cohort design disease-causing mutation experimental study frontal lobe frontotemporal lobar dementia amyotrophic lateral sclerosis fused in sarcoma in vivo innovation insight interdisciplinary approach mRNA Decay mRNA Surveillance mouse model mutant neuron loss neurotoxicity nonalzheimer dementia novel optogenetics polysome profiling protein aggregation spatiotemporal therapeutic development transcriptome sequencing translational impact

项目摘要

Frontotemporal dementia (FTD) is an age-related non-Alzheimer dementia characterized by progressive neuronal loss in the frontotemporal lobes. A subset of FTD is defined by the pathology of protein inclusions positive for Fused in Sarcoma (FUS), thus named FUS-FTD. FTD and amyotrophic lateral sclerosis (ALS) share a wide spectrum of clinical, pathological and genetic features. Pathogenic mutations of FUS cause both ALS and FTD, and FUS proteinopathy is also detected in sporadic diseases. FUS is primarily in the nucleus, but the protein with a disease-causing mutation mislocalizes and accumulates in the cytoplasm. Protein aggregates induced by FUS mutations may sequester other proteins to compromise cellular functions, resulting in impaired neurons. We recently showed that FTD/ALS mutations of FUS suppressed protein translation and hyper-activated the nonsense-mediated decay (NMD) of mRNAs. The hypothesis to be tested in this project is that the dysregulation of protein translation and mRNA surveillance contributes to cortical neuron loss in FUS-FTD. Three specific aims are designed to test the hypothesis using in vitro and animal models as well as FTD patient tissues. Aim 1 is to determine how mRNA NMD and protein translation are perturbed by pathogeneic FUS mutations in FTD mouse models and patient tissues. We will determine whether NMD factors and translation-related proteins are sequestered in FUS inclusions in forebrain neurons in R521G FUS transgenic mice at different ages. We will measure mRNA turnover rates and protein translation efficiency in R521G FUS mice and correlate the perturbations to neuronal dysfunction and FTD disease progression. Moreover, we will examine whether NMD factors and protein translation proteins are sequestered in FTD patient tissues. Aim 2 is to identify specific proteins suppressed by pathogenic FUS in FTD mice. We will apply the puromycin labeling in FTD mice and use the proteomic approach to identify changes in protein translation impacted by mutant FUS in forebrain neurons. In addition, actively translated mRNAs will be identified and quantified in polysome fractions using RNA-Seq. Results from the –omics approaches will be integrated for pathway analysis to reveal whether pathogenic FUS impairs proteins in specific pathways, providing novel insights into the FTD etiology. Aim 3 is to elucidate the significance of RNA binding and post-translational modifications in the dysfunction of pathogenic FUS. We will use a cohort of RNA binding-deficient mutations in an optogenetic Cry2olig-FUS-mCherry system to examine the significance of RNA binding in FUS inclusion formation and dysfunction in cortical neurons. We recently found FUS is acetylated at residues critical to RNA binding, we will test how acetylation-null and -mimicking mutations affect FUS inclusions, NMD and protein translation. In addition, we will also examine whether pathogenic FUS forms RNA-dependent and -independent inclusions that produce different levels of toxicity to neurons. The proposed experiments will thoroughly examine a novel disease mechanism using innovative approaches. Completion of our proposed work will help elucidate molecular mechanisms underlying FUS FTD.

额颞痴呆（FTD）是与年龄相关的非阿尔茨海默氏痴呆症，其特征是进行性。额颞爱的神经元损失。 FTD的子集由蛋白质夹杂物的病理定义在肉瘤（FUS）中融合的阳性，因此被称为fus-ftd。 FTD和肌萎缩性侧面硬化症（ALS）共享广泛的临床，病理和遗传特征。 FUS的致病突变引起ALS和在零星疾病中还检测到FTD和FUS蛋白质病。 FUS主要在细胞核中，但是引起疾病的突变的蛋白质会误置并积聚在细胞质中。蛋白质聚集体通过FUS突变诱导的可能会隔离其他蛋白质，以损害细胞功能，从而受损神经元。我们最近表明，FUS的FTD/ALS突变抑制了蛋白质翻译，并过度激活了mRNA的胡说八道介导的衰减（NMD）。在这个项目中要检验的假设是蛋白质翻译和mRNA监测的失调会导致FUS-FTD的皮质神经元丧失。设计了三个特定目标，用于使用体外和动物模型以及FTD检验假设患者组织。 AIM 1是确定mRNA NMD和蛋白质翻译如何受到致病性FUS的干扰 FTD小鼠模型和患者组织中的突变。我们将确定在R521g FUS转基因小鼠的前脑神经元中，NMD因子和翻译相关的蛋白是否在FUS夹杂物中隔离不同的年龄。我们将测量R521G FUS小鼠的mRNA转换率和蛋白质翻译效率，并将扰动与神经元功能障碍和FTD疾病进展相关。而且，我们将检查 NMD因子和蛋白质翻译蛋白是否在FTD患者组织中隔离。目标2是确定在FTD小鼠中被致病性FU抑制的特异性蛋白质。我们将在FTD小鼠中施加嘌呤霉素标记并使用蛋白质组学方法来识别前脑突变FUS影响的蛋白质翻译的变化神经元。此外，将使用RNA-Seq在多质体组中鉴定并定量积极翻译的mRNA。 –omics方法的结果将集成以进行途径分析，以揭示病原 FUS会损害特定途径中的蛋白质，从而为FTD病因提供新的见解。目标3是阐明 RNA结合和翻译后修饰在致病性FUS功能障碍中的重要性。我们将在光遗传学cry2olig-fus-mcherry系统中使用一系列RNA结合缺陷突变来检查 RNA结合在皮质神经元中FUS纳入形成和功能障碍中的重要性。我们最近发现FUS在对RNA结合至关重要的残留物处是乙酰化的，我们将测试乙酰化和仿真的方式突变影响FUS夹杂物，NMD和蛋白质翻译。此外，我们还将检查是否致病性FU形成RNA依赖性和非依赖性包含物，产生不同水平的毒性神经元。拟议的实验将使用创新的新型疾病机制进行彻底检查方法。我们提出的工作的完成将有助于阐明FUS FUS FTD的分子机制。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Frontotemporal dementia non-sense mutation of progranulin rescued by aminoglycosides.

氨基糖甙类药物挽救的额颞叶痴呆颗粒体蛋白前体无义突变。

DOI：
10.1093/hmg/ddz280
发表时间：
2020
期刊：
Human molecular genetics
影响因子：
3.5
作者：
Kuang,Lisha;Hashimoto,Kei;Huang,EricJ;Gentry,MatthewS;Zhu,Haining
通讯作者：
Zhu,Haining

Conditional depletion of Fus in oligodendrocytes leads to motor hyperactivity and increased myelin deposition associated with Akt and cholesterol activation.

DOI：
10.1002/glia.23825
发表时间：
2020-10
期刊：
Glia
影响因子：
6.2
作者：
Guzman KM;Brink LE;Rodriguez-Bey G;Bodnar RJ;Kuang L;Xing B;Sullivan M;Park HJ;Koppes E;Zhu H;Padiath Q;Cambi F
通讯作者：
Cambi F