Single cell analysis of the kinome

项目摘要 分析激酶及其在单细胞中的活性将使得能够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 在过去的十年中，通过单细胞基因组学，我们仍然了解不同细胞类型的状态， 对蛋白质，特别是不同细胞群体中的激酶网络的研究很少。这主要是 因为单细胞激酶组分析面临重大的技术挑战。与核酸不同， 蛋白质不能被扩增，使得从单个细胞中检测微量蛋白质变得困难。我们 最近展示了一个原理验证实验，以准确检测蛋白质PTM亚型 在单个细胞中。在这里，我们建议将这种方法扩展到全球水平的蛋白激酶， 在细胞培养物和组织中的单个细胞。我们将整合单细胞激酶组分析 通过RNA测量来鉴定细胞类型特异性激酶谱。最终，我们将绘制 在小鼠和人脑中具有空间背景的激酶组，以检查激酶通路 与衰老和神经退化有关，可以确定药物靶点。这个项目是一个 这与我们目前的研究有很大的不同。它的目标雄心勃勃，但可以实现。项目 我们将利用我们在单细胞技术开发和新技术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.