Simultaneous Multiplexed Measurement of RNA and Proteins in Single Cells.
Simultaneous Multiplexed Measurement of RNA and Proteins in Single Cells.
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DOI：
10.1016/j.celrep.2015.12.021
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发表时间：
2016-01-12
期刊：
影响因子：
8.8
通讯作者：
Landegren U
中科院分区：
文献类型：
--
作者：
Darmanis S;Gallant CJ;Marinescu VD;Niklasson M;Segerman A;Flamourakis G;Fredriksson S;Assarsson E;Lundberg M;Nelander S;Westermark B;Landegren U
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，但要完全定义细胞状态，还必须将蛋白质作为定义细胞表型的核心因素。目前用于分析单细胞中内源性蛋白质表达的方法在特异性、通量或多重能力方面受到限制。在这里，我们提出了一种方法，同时和具体询问大组蛋白质和RNA的目标在裂解物从个别细胞，使细胞功能和反应的调查。我们应用我们的方法来研究BMP 4（一种实验性治疗剂）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和蛋白质水平上对治疗的反应具有显著的异质性，其中一部分细胞以不同的方式对BMP 4反应。此外，我们发现蛋白质和RNA在单细胞水平上的相关性总体较差，蛋白质更准确地定义了对治疗的反应。单细胞中的多重蛋白质和RNA测量从RNA和蛋白质数据定义细胞状态的独特信息胶质母细胞瘤细胞中对推定的治疗性BMP 4的反应的异质性Darmanis et al.提出了一种方法，可以同时测量单细胞中多达96种转录物和蛋白质的水平。他们应用这种技术来研究胶质母细胞瘤细胞对BMP 4（一种拟议的治疗剂）的反应，并揭示异质性反应和细胞状态。
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. Multiplexed protein and RNA measurement in single cells Distinct information for defining cell states from RNA and protein data Heterogeneity of responses to the putative therapeutic BMP4 in glioblastoma cells Darmanis et al. present an approach to simultaneously measure levels of up to 96 transcripts and proteins in single cells. They apply this technique to study responses of glioblastoma cells to BMP4, a proposed therapeutic agent, and reveal heterogeneous responses and cell states.