Dissecting neuron-microglia-astrocyte interaction in AD pathogenesis

项目摘要/摘要 这个项目的长期目标是了解与年龄相关的神经元-小胶质细胞-星形胶质细胞特有的蛋白质组。 阿尔茨海默病(AD)发病机制中脑内的动力学和相互作用以及它们的失控。 我们正在进行的大规模AD脑组织转录组和蛋白质组图谱显示， 转录组通常不是蛋白质丰度的准确指标，因为转录后事件，如 蛋白质周转率。遗传学研究已经发现了20多个与AD相关的基因，其中许多与 特定的细胞类型。我们假设神经元-小胶质细胞-星形胶质细胞相互作用功能障碍与AD有关 发病机制，并建议使用最新的尖端技术来描述细胞类型特定的蛋白质组，在 多种常见AD相关小鼠三种细胞群的原位转录组和蛋白质组转换 模型，破译疾病发展中的分子信号网络，并验证人类AD的靶点 病例、小鼠模型和人体器官模型。三个具体目标是：(I)解开动态神经元-- BONCAT介导的单细胞型AD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)测量阿尔茨海默病细胞类型特异性蛋白质组全蛋白周转 通过BONCAT脉冲追逐分析，以及(Iii)研究人类AD病例中蛋白质组动力学的放松调控， 小鼠模型和脑器官。该项目的成功结果将确定新的、特定的细胞类型 AD发病过程中调节细胞-细胞相互作用的分子成分和途径，可能 为有效治疗这种破坏性疾病提供新的治疗策略。

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.