Multiplexed mass cytometry profiling of cellular states perturbed by small-molecule regulators

• Published in: Nature biotechnology Vol. 30; no. 9; pp. 858 - 867
• Main Authors: Bodenmiller, Bernd, Zunder, Eli R, Finck, Rachel, Chen, Tiffany J, Savig, Erica S, Bruggner, Robert V, Simonds, Erin F, Bendall, Sean C, Sachs, Karen, Krutzik, Peter O, Nolan, Garry P
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01.09.2012
• Subjects: Cellular biology; Chelating Agents; Flow cytometry; Flow Cytometry - methods; Heterocyclic Compounds, 1-Ring; High-Throughput Screening Assays - methods; Humans; K562 Cells; Leukocytes, Mononuclear - cytology; Leukocytes, Mononuclear - drug effects; Leukocytes, Mononuclear - metabolism; Methods; Molecular dynamics; Molecules; Phosphorylation; Principal Component Analysis; Protein Kinase Inhibitors - pharmacology; Signal Transduction; Systems Biology - methods; United States
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt.2317

Mass cytometry facilitates high-dimensional, quantitative analysis of the effects of bioactive molecules on human samples at single-cell resolution, but instruments process only one sample at a time. Here we describe mass-tag cellular barcoding (MCB), which increases mass cytometry throughput by using n metal ion tags to multiplex up to 2n samples. We used seven tags to multiplex an entire 96-well plate, and applied MCB to characterize human peripheral blood mononuclear cell (PBMC) signaling dynamics and cell-to-cell communication, signaling variability between PBMCs from eight human donors, and the effects of 27 inhibitors on this system. For each inhibitor, we measured 14 phosphorylation sites in 14 PBMC types at 96 conditions, resulting in 18,816 quantified phosphorylation levels from each multiplexed sample. This high-dimensional, systems-level inquiry allowed analysis across cell-type and signaling space, reclassified inhibitors and revealed off-target effects. High-content, high-throughput screening with MCB should be useful for drug discovery, preclinical testing and mechanistic investigation of human disease.