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

Pron病是一种罕见的、总是致命的神经退行性疾病，其特征是迅速 认知和运动能力下降。某些病理特征与阿尔茨海默病重叠， 包括蛋白质聚集和大脑中大量突触的丢失，但事件的顺序 导致突触丢失的原因还不完全清楚。在普恩和阿尔茨海默病模型中， 据报道，神经细胞蛋白(PrPC)与寡聚体结合并引起一连串的 细胞内信号，而PrPC缺失可以改善突触损伤，这强烈表明 神经元PrPC在改变信号转导事件中的作用。利用无偏转录学技术在普里恩上- 感染小鼠大脑后，我们发现即刻早期的基因是最早、最多的 显著上调了海马体中的基因，提示神经元活动增强。 我们还在海马区和大脑皮层发现了早期异常的神经元激活酶活性。 值得注意的是，在人ipsc诱导的神经元中，在2小时内也激发了类似的激酶修饰。 触发PrPC的可能性。在目标1中，我们将确定抗PrP抗体如何配基或纯化 传染性Pron寡聚体在人iPSC诱导的神经元中诱导突触激酶信号转导。在……里面 目的2.利用高灵敏的定量蛋白质组学方法鉴定PrPC跨膜蛋白 普恩蛋白引发的人脑细胞信号网络组成和网络改变 蛋白质配基。这些研究首次对PrPC启动的人类信号通路进行了研究 神经元，并有望建立信号参数和下游后果 以及对疾病中导致突触丢失的事件的洞察。

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.