Dissecting neuron-microglia-astrocyte interaction in AD pathogenesis

项目概要/摘要 这个项目的长期目标是了解年龄相关的、神经元-小胶质细胞-星形胶质细胞特异性蛋白质组 动态和相互作用的大脑和他们的失调的发病机制阿尔茨海默病（AD）。 我们正在进行的大量AD脑组织中转录组和蛋白质组的大规模分析表明， 由于转录后事件，转录组通常不是蛋白质丰度的准确指示物，例如 蛋白质周转遗传学研究已经揭示了超过20个AD相关基因，其中许多与AD相关。 特定细胞类型。我们推测，神经元-小胶质细胞-星形胶质细胞相互作用的功能障碍有助于AD 发病机制，并建议使用最新的尖端技术来分析细胞类型特异性蛋白质组， 多个常见AD相关小鼠的三个细胞群中的原位转录组和蛋白质组更新 模型，以破译疾病发展中的分子信号网络，并验证人类AD中的靶点 病例、小鼠模型和人类类器官模型。三个具体目标是：（一）解开动态神经元- 通过BONCAT介导的单细胞型AD小鼠小胶质细胞-星形胶质细胞特异性蛋白质组及其相互作用 蛋白质组学和空间转录组学，（ii）测量AD中细胞类型特异性蛋白质组范围内的蛋白质周转 通过BONCAT脉冲追踪分析，以及（iii）研究人类AD病例中蛋白质组动力学的失调， 小鼠模型和脑类器官。该项目的成功结果将确定新的，细胞类型特异性 AD发病过程中调节细胞-细胞相互作用的分子组分和途径， 为有效治疗这种毁灭性疾病提供了新的治疗策略。

PROJECT SUMMARY/ABSTRACT The long-term goal of this project is to understand the age-related, neuron-microglia-astrocyte-specific proteome dynamics and interaction in the brain and their deregulation in the pathogenesis of Alzheimer’s disease (AD). Our ongoing large-scale profiling of transcriptome and proteome in bulk AD brain tissue have revealed that the transcriptome is often not an accurate indicator of protein abundance, due to posttranscriptional events, such as protein turnover. Genetic studies have revealed over 20 AD-related genes, many of which are associated with specific cell types. We hypothesize that dysfunction of neuron-microglia-astrocyte interaction contributes to AD pathogenesis, and propose to use the latest cutting edge technologies to profile cell type specific proteome, in situ transcriptome, and proteome turnover in the three cell populations from multiple common AD-related mouse models, to decipher molecular signaling networks in disease development, and to validate targets in human AD cases, mouse models, and human organoid models. Three specific aims are: (i) to disentangle dynamic neuron- microglia-astrocyte-specific proteome and interaction in AD mice by BONCAT-mediated single cell type proteomics and spatial transcriptomics, (ii) to measure cell type-specific proteome-wide protein turnover in AD by BONCAT pulse-chase analysis, and (iii) to study deregulation of proteome dynamics in human AD cases, mouse models and brain organoids. Successful outcome of this project will identify novel, cell type specific molecular components and pathways regulating cell-cell interactions during AD pathogenesis, which may provide new therapeutic strategies for effective treatment of this devastating disorder.