Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala 75108, Sweden; Science for Life Laboratory, Uppsala University, Uppsala 75108, Sweden.
Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala 75108, Sweden.
Cell Rep. 2016 Jan 12;14(2):380-9. doi: 10.1016/j.celrep.2015.12.021. Epub 2015 Dec 31.
Significant advances have been made in methods to analyze genomes and transcriptomes of single cells, but to fully define cell states, proteins must also be accessed as central actors defining a cell's phenotype. Methods currently used to analyze endogenous protein expression in single cells are limited in specificity, throughput, or multiplex capability. Here, we present an approach to simultaneously and specifically interrogate large sets of protein and RNA targets in lysates from individual cells, enabling investigations of cell functions and responses. We applied our method to investigate the effects of BMP4, an experimental therapeutic agent, on early-passage glioblastoma cell cultures. We uncovered significant heterogeneity in responses to treatment at levels of RNA and protein, with a subset of cells reacting in a distinct manner to BMP4. Moreover, we found overall poor correlation between protein and RNA at the level of single cells, with proteins more accurately defining responses to treatment.
在分析单细胞基因组和转录组的方法方面已经取得了重大进展，但为了全面定义细胞状态，还必须将蛋白质作为定义细胞表型的核心因素来进行研究。目前用于分析单细胞内源性蛋白质表达的方法在特异性、通量或多重检测能力方面存在局限性。在这里，我们提出了一种在单个细胞裂解物中同时且特异性检测大量蛋白质和 RNA 靶标的方法，从而能够研究细胞功能和反应。我们应用该方法研究了 BMP4（一种实验性治疗药物）对早期传代胶质母细胞瘤细胞培养物的影响。我们发现，在 RNA 和蛋白质水平上，细胞对治疗的反应存在显著异质性，一小部分细胞对 BMP4 的反应明显不同。此外，我们还发现单细胞水平上蛋白质和 RNA 之间的整体相关性较差，蛋白质更能准确地定义对治疗的反应。
