Determining pathogenic PrPC-induced signaling pathways in human iPSC-induced neurons

确定人 iPSC 诱导神经元中致病性 PrPC 诱导的信号通路

基本信息

批准号：
10791127
负责人：
Christina Sigurdson
金额：
$ 43.45万
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：
2023
资助国家：
美国
起止时间：
2023-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10791127
关键词：
AMPA Receptors Acute Affect Alzheimer&apos s Disease Alzheimer&apos s disease model Antibodies Automobile Driving Binding Binding Proteins Brain Calcium Calcium Signaling Calmodulin Chronic Cognitive Complex DAP kinase Data Disease Dissection Early identification Enzymes Epilepsy Epitopes Event Functional disorder Gene Expression Genes Glutamate Receptor Goals Hippocampus Hour Human Immediate-Early Genes Immunoprecipitation Impairment In Vitro Infection Integral Membrane Protein Ligands Link Mass Spectrum Analysis Mediating Membrane Membrane Microdomains Membrane Proteins Modeling Modification Motor Mus Neurodegenerative Disorders Neurons Pathogenicity Pathologic Pathway interactions Phosphorylation Phosphotransferases Physiological Postsynaptic Membrane PrP PrPSc Proteins Prion Diseases Prions Process Protein Kinase Protein-Serine-Threonine Kinases Proteins Proteome Proteomics Rare Diseases Receptor Signaling Regulation Reporting Role STEM research Serine Signal Induction Signal Pathway Signal Transduction Signaling Molecule Signaling Protein Stroke Structure Surface Synapses Synaptic Membranes Synaptic plasticity Techniques abeta oligomer anti-PrP antibodies calmodulin-dependent protein kinase II curative treatments excitotoxicity glutamatergic signaling in vivo induced pluripotent stem cell insight link protein neuron loss new therapeutic target phosphoproteomics postsynaptic protein activation protein aggregation protein complex receptor release of sequestered calcium ion into cytoplasm response synaptic function trafficking transcriptomics

项目摘要

Prion diseases are rare, invariably fatal neurodegenerative disorders characterized by rapid cognitive and motor decline. Certain pathologic features overlap with Alzheimer’s disease, including protein aggregates and massive synapse loss in the brain, yet the sequence of events driving synaptic loss is incompletely understood. In prion and Alzheimer’s disease models, neuronal cellular prion protein (PrPC) reportedly binds oligomers and elicits a cascade of intracellular signals, whereas PrPC deletion ameliorates synaptic impairment, strongly implicating neuronal PrPC in altering signal transduction events. Using unbiased transcriptomics on prion- infected mouse brain, we have found that immediate early genes are among the earliest, most significantly upregulated genes in the hippocampus, suggestive of heightened neuronal activity. We have also discovered early aberrant neuronal kinase activity in the hippocampus and cortex. Notably, similar kinase modifications were incited in human iPSC-induced neurons within 2 hours of triggering PrPC. In Aim 1, we will determine how an anti-PrP antibody ligand or purified infectious prion oligomers induce synaptic kinase signaling in human iPSC-induced neurons. In Aim 2, we use highly sensitive and quantitative proteomics to identify the PrPC- transmembrane signaling network components and network alterations in human neurons triggered by a prion protein ligand. These studies are the first to pursue PrPC-initiated signaling pathways in human neurons, and are expected to establish the signaling parameters and downstream consequences as well as provide insight into events leading to synapse loss in disease.

朊病毒病是罕见的、总是致命的神经退行性疾病，其特征是快速认知和运动能力下降。某些病理特征与阿尔茨海默病重叠，包括大脑中的蛋白质聚集和大量突触损失，但事件的顺序在朊病毒和阿尔茨海默病模型中，驱动突触损失的机制尚不完全清楚。据报道，神经元细胞朊病毒蛋白 (PrPC) 与寡聚体结合并引发一系列级联反应细胞内信号，而 PrPC 缺失可改善突触损伤，强烈暗示神经元 PrPC 改变信号转导事件。感染小鼠大脑后，我们发现立即早期基因是最早、最多的基因之一海马体中的基因显着上调，表明呼吸神经元活动。我们还发现海马体和皮质中早期异常的神经元激酶活性。值得注意的是，2 小时内在人类 iPSC 诱导的神经元中诱导了类似的激酶修饰在目标 1 中，我们将确定如何配体或纯化抗 PrP 抗体。传染性朊病毒寡聚体在人类 iPSC 诱导的神经元中诱导突触激酶信号传导。目标2，我们使用高灵敏度和定量的蛋白质组学来鉴定PrPC-跨膜朊病毒触发的人类神经元信号网络组件和网络改变这些研究首次在人类中探索 PrPC 启动的信号通路。神经元，并有望建立信号参数和下游后果以及提供对导致疾病中突触丢失的事件的深入了解。