Molecular basis of prion protein-induced neurodegeneration

朊病毒蛋白诱导神经变性的分子基础

基本信息

批准号：
10199633
负责人：
Christina Sigurdson
金额：
$ 163.4万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
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：
2021-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10199633
关键词：
Affect Age Alzheimer&apos s Disease Alzheimer&apos s disease model Amino Acid Substitution Amyloid beta-Protein Animals Automobile Driving Autophagosome Binding Biochemical Brain Brain Diseases Calcium Signaling Chronic Clinical Creutzfeldt-Jakob Syndrome Data Deposition Disease Dissection Engineering Extracellular Protein Functional disorder Gliosis Glutamate Receptor Glutamates Glycoproteins Goals Health Histologic Human Impairment In Vitro Infection Knock-in Knock-in Mouse Link Location Maintenance Mass Spectrum Analysis Mediating Modeling Molecular Molecular Conformation Morphology Mus N-terminal Nerve Degeneration Neurites Neurodegenerative Disorders Neurologic Signs Neurons Outcome Pathologic Pathology Pathway interactions Patients Phosphorylation Point Mutation Polysaccharides Post-Translational Protein Processing PrP PrP gene PrPSc Proteins Prion Diseases Prions Proteins Proteomics Rare Diseases Reactive Oxygen Species Role Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Structure Suggestion Synapses Synaptic Membranes Synaptic Receptors Synaptosomes Testing Toxic effect Transgenic Mice Vacuole Variant Varicosity alpha synuclein base bone curative treatments excitotoxicity in vivo insight mouse model mutant neuron loss neurotoxic neurotoxicity neurotransmission new therapeutic target protein aggregation protein expression protein function protein structure proteostasis receptor function receptor sensitivity response synaptic function tau aggregation therapeutic development trafficking vesicle transport

项目摘要

Prion diseases are rare, invariably fatal neurodegenerative disorders with pathologic features in common with Alzheimer’s disease, including extracellular protein aggregates, synaptic loss, and neuritic dystrophy. In prion and Alzheimer’s disease models, depletion of neuronal cellular prion protein (PrPC) ameliorates synaptic impairment and clinical disease, strongly implicating neuronal PrPC expression in the altered signal transduction cascades that may underlie synaptotoxicity and endolysosomal dysfunction. We have engineered the first knock-in mouse model with a point mutation in Prnp that develops a striking and severe spongiform encephalopathy, neuritic dystrophy, and altered post-synaptic receptor phosphorylation, in the absence of prion aggregates. Cultured cortical neurons from these knock-in mice show an increased sensitivity to glutamate and dendritic varicosities, suggestive of excitotoxicity. Thus, this PrP knock-in model provides a unique opportunity to elucidate key PrPC interactions and altered signal transduction pathways at the synapse and to determine the molecular mechanisms that link PrPC to synaptic loss and endolysosomal dysregulation. Our long-term goal is to understand how PrPC triggers aberrant neuronal signaling that may drive impaired proteostasis and synaptotoxicity in prion disease. Using cultured primary neurons and mice, we will first determine how the mutant PrPC interactions impact pre- and post-synaptic neuronal protein levels and glutamate receptor function. We will then identify how mutant PrPC dysregulates endolysosomal and proteostatic activity. Finally, we use highly sensitive and quantitative proteomics to define the PrP interactome and phosphoproteome network alterations in the brain by tandem mass tag mass spectrometry analysis. For all aims, we will directly test how the findings from the mutant PrPC-expressing brain compare to prion-infected mouse and human brain. These studies are the first to target the neuronal endolysosomal and synaptic pathways in a knock-in mouse model expressing mutant PrPC, and outcomes are expected to provide key insights into the role of PrPC in synapse maintenance and the signaling pathways inciting synaptic loss, thus revealing new therapeutic targets for prion disease.

prion病是罕见的，始终具有致命的神经退行性疾病，具有病理特征患有阿尔茨海默氏病的常见，包括细胞外蛋白质骨料，突触损失和神经性营养不良。在prion和阿尔茨海默氏病模型中，神经元细胞幼虫的耗竭蛋白质（PRPC）可以改善合成障碍和临床疾病，强烈暗示在改变的信号转导级联反应中的神经元PRPC表达可能是基础的突触毒性和内溶性功能障碍。我们已经设计了第一个敲门鼠标 PRNP中具有点突变的模型，该模型会发展出罢工和严重的赞助形式脑病，神经性营养不良症和突触后受体磷酸化改变，在缺乏prion骨聚集体。这些敲门小鼠的培养的皮质神经元显示提高对谷氨酸和树突状静脉曲张的敏感性，提示兴奋性毒性。那，这种PRP敲击模型为阐明关键PRPC互动的独特机会和改变突触处的信号转移途径并确定分子将PRPC连接到突触损失和内溶性失调的机制。我们的长期目标是了解PRPC如何触发可能驱动受损的异常神经元信号蛋白质毒性和突触毒性。使用培养的原代神经元和小鼠，我们将首先确定突变体PRPC相互作用如何影响突触后神经元前后蛋白水平和谷氨酸受体功能。然后，我们将确定突变的PRPC 失调的内溶液体和蛋白抑制活性。最后，我们使用高度敏感的定量蛋白质组学来定义PRP Interactome和磷蛋白组网络的变化通过串联质量质谱分析在大脑中。对于所有目标，我们将直接测试表达突变体PRPC的大脑的发现与原始的小鼠相比如何人脑。这些研究是第一个靶向神经元内溶血和突触的研究表达突变体PRPC的敲击小鼠模型中的途径，预期结果将为提供有关PRPC在突触维持和信号通路中的作用的关键见解煽动突触损失，从而揭示了针对Prion疾病的新治疗靶标。