TY - JOUR T1 - Single-Cell Signalling Analysis of Heterocellular Organoids JF - bioRxiv DO - 10.1101/659896 SP - 659896 AU - Xiao Qin AU - Jahangir Sufi AU - Petra Vlckova AU - Pelagia Kyriakidou AU - Sophie E. Acton AU - Vivian S. W. Li AU - Mark Nitz AU - Christopher J. Tape Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/06/06/659896.abstract N2 - Organoids are biomimetic 3D models of healthy and diseased tissue. Despite their widespread adoption, methods to analyse cell-type specific signalling networks in organoids are absent. Here we report multiplexed single-cell analysis of post-translational modification (PTM) signalling networks in organoids by mass cytometry. Simultaneous analysis of 28 PTMs in >1 million single-organoid cells reveals cell-type and cell-state specific signalling networks in stem, Paneth, enteroen-docrine, goblet, tuft cells, and enterocytes during intestinal organoid development. We demonstrate that Thiol-reactive Organoid Barcoding in situ (TOBis) enables high-throughput multiplexed organoid signalling analysis in a single-tube – opening this technology to single-cell organoid screening. Comparison of colorectal cancer (CRC) oncogenic mutations (Apc, Kras, and Trp53) and tumour microenvironmental cues (fibroblasts and macrophages) revealed CRC driver-mutations mimic signalling normally provided by stromal cells. These results demonstrate mass cytometry is a powerful multiplexed single-cell technology for studying cell-specific signalling in organoid models of healthy and cancerous tissue. ER -