Institute for Molecules and Materials, Radboud University, Nijmegen 6525 AJ, the Netherlands.
Aduro Biotech, Oss 5349 AB, the Netherlands.
Cell Rep Methods. 2021 Sep 15;1(5):100070. doi: 10.1016/j.crmeth.2021.100070. eCollection 2021 Sep 27.
To further our understanding of how biochemical information flows through cells upon external stimulation, we require single-cell multi-omics methods that concurrently map changes in (phospho)protein levels across signaling networks and the associated gene expression profiles. Here, we present quantification of RNA and intracellular epitopes by sequencing (QuRIE-seq), a droplet-based platform for single-cell RNA and intra- and extracellular (phospho)protein quantification through sequencing. We applied QuRIE-seq to quantify cell-state changes at both the signaling and the transcriptome level after 2-, 4-, 6-, 60-, and 180-min stimulation of the B cell receptor pathway in Burkitt lymphoma cells. Using the multi-omics factor analysis (MOFA+) framework, we delineated changes in single-cell (phospho)protein and gene expression patterns over multiple timescales and revealed the effect of an inhibitory drug (ibrutinib) on signaling and gene expression landscapes.
为了进一步了解细胞在外部刺激下的生化信息如何传递,我们需要单细胞多组学方法,能够同时绘制信号网络中(磷酸化)蛋白水平的变化以及相关的基因表达谱。在这里,我们提出了通过测序进行 RNA 和细胞内表位定量(QuRIE-seq),这是一种基于液滴的单细胞 RNA 以及细胞内和细胞外(磷酸化)蛋白定量的测序平台。我们应用 QuRIE-seq 来量化 Burkitt 淋巴瘤细胞中 B 细胞受体通路在 2、4、6、60 和 180 分钟刺激后在信号和转录组水平上的细胞状态变化。使用多组学因子分析(MOFA+)框架,我们描绘了单细胞(磷酸化)蛋白和基因表达模式在多个时间尺度上的变化,并揭示了一种抑制性药物(依鲁替尼)对信号和基因表达图谱的影响。