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bioRxiv · 10.1101/659896

Single-Cell Signalling Analysis of Heterocellular Organoids

Qin, X.·Sufi, J.·Vlckova, P.·Kyriakidou, P.·Acton, S. E.·Li, V. S. W.·Nitz, M.·Tape, C. J.
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Abstract

Organoids are powerful biomimetic tissue models. Despite their widespread adoption, methods to analyse cell-type specific post-translational modification (PTM) signalling networks in organoids are absent. Here we report multivariate single-cell analysis of cell-type specific signalling networks in organoids and organoid co-cultures. Simultaneous measurement of 28 PTMs in >1 million single small intestinal organoid cells by mass cytometry reveals cell-type and cell-state specific signalling networks in stem, Paneth, enteroendocrine, tuft, goblet cells, and enterocytes. Integrating single-cell PTM analysis with Thiol-reactive Organoid Barcoding in situ (TOBis) enables high-throughput comparison of signalling networks between organoid cultures. Multivariate cell-type specific PTM analysis of colorectal cancer tumour microenvironment organoids reveals that shApc, KrasG12D, and Trp53R172H cell-autonomously mimic signalling states normally induced by stromal fibroblasts and macrophages. These results demonstrate how standard mass cytometry workflows can be modified to perform high-throughput multivariate cell-type specific signalling analysis of healthy and cancerous organoids.

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Qin, X., Sufi, J., Vlckova, P., Kyriakidou, P., Acton, S. E., Li, V. S. W., Nitz, M., Tape, C. J.. 2019-06-06. Single-Cell Signalling Analysis of Heterocellular Organoids. https://doi.org/10.1101/659896

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