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.

病毒疾病很少见，始终是致命的神经退行性疾病，其特征是快速认知和运动下降。某些病理特征与阿尔茨海默氏病重叠，包括蛋白质骨料和大脑中的大规模突触丧失，但事件的序列驾驶合成损失尚不完全理解。在王室和阿尔茨海默氏病模型中据报道，神经元细胞prion蛋白（PRPC）结合低聚物并引起级联细胞内信号，而PRPC缺失可以改善突触障碍，强烈暗示神经元PRPC改变了信号转导事件。在prion上使用公正的转录组学 - 感染的小鼠大脑，我们发现早期基因立即是最早的基因，大多数是海马中明显上调基因，表明神经元活性的增强。我们还发现了海马和皮质中的早期异常神经元激酶活性。值得注意的是，在2小时内，在人IPSC诱导的神经元中煽动了类似的激酶修饰触发PRPC。在AIM 1中，我们将确定抗PRP抗体配体或纯化传染性的prion低聚物诱导人IPSC诱导的神经元中的突触激酶信号传导。 AIM 2，我们使用高度敏感和定量的蛋白质组学来识别PRPC-跨膜由素数触发的人类神经元中的信号网络组件和网络变化蛋白质配体。这些研究是第一个追求人类PRPC发起的信号通路的研究神经元，并有望建立信号传导参数和下游后果并提供有关导致疾病突触丧失的事件的见解。