Single cell analysis of the kinome

项目概要 分析单细胞中的激酶及其活性将有助于探索细胞类型特异性 功能和生化途径。虽然我们已经了解了很多关于转录的知识 通过单细胞基因组学，我们仍然了解过去十年不同细胞类型的状态 关于不同细胞群中的蛋白质，特别是激酶网络的知识很少。这很大程度上是 因为单细胞激酶组分析面临重大技术挑战。与核酸不同， 蛋白质无法被扩增，使得从单细胞中检测微量蛋白质变得困难。我们 最近展示了一项原理验证实验，可准确检测蛋白质 PTM 亚型 在单细胞中。在这里，我们建议将该方法扩展到全球范围内的激酶分析 细胞培养物和组织中的单细胞。我们将整合单细胞激酶组分析 通过 RNA 测量来识别细胞类型特异性激酶谱。最终我们将绘制地图 小鼠和人脑中具有空间背景的激酶组以检查激酶途径 参与衰老和神经退行性变，可以识别可药物靶标。这个项目是一个 与我们当前研究的重大背离。它的目标雄心勃勃但可以实现。项目 将利用我们在单细胞技术开发和新应用方面的专业知识 技术应用于不同的生物系统。

Project Summary Profiling kinases and their activities in single cells will enable exploration of cell type specific functional and biochemical pathways. While we have learned a great deal about transcriptional states of different cell types in the past decade through single cell genomics, we still understand little about proteins and in particular kinases networks in diverse cell populations. This is largely because single-cell kinome analysis faces major technical challenges. Unlike nucleic acids, proteins cannot be amplified, making detection of minute quantities from single cells difficult. We recently demonstrated a proof-of-principle experiment to accurately detect protein PTM isoforms in single cells. Here we propose to scale this method up to profile kinases at the global level in single cells in both cell culture and in tissues. We will integrate the single cell kinome analysis with RNA measurements to identify cell type specific kinase profiles. Ultimately, we will map the kinome with spatial context in the mouse and human brain to examine the kinase pathways involved in aging and neurodegeneration which could identify druggable targets. This project is a major departure from our current research. Its goals are ambitious but achievable. The project will leverage our expertise in single cell technology development and in application of new technology to diverse biological systems.