Single-Cell Signalling Analysis of Tumour Microenvironment Organoids
Abstract
Organoids are self-organising stem cell-derived ex vivo cultures widely adopted as biomimetic models of healthy and diseased tissues. As complex heterocellular systems, organoids are especially well-positioned to take advantage of emerging high-dimensional single-cell technologies. Here we present a Cytometry by Time-Of-Flight (CyTOF) method for single-cell analysis of post-translational modification (PTM) signalling in organoids and tumour microenvironment organoid co-cultures. Integrating single-cell PTM analysis with thiol-reactive organoid barcoding in situ (TOBis) enables 35-plex and 126-plex comparison of signalling networks between organoid co-cultures. Cell-type-specific PTM analysis of colorectal cancer organoid co-cultures revealed that oncogenic mutations cell-autonomously mimic signalling states normally induced by stromal fibroblasts and macrophages.
Biosketch
Chris received his Ph.D. from Prof. Gillian Murphy’s lab at the CRUK Cambridge Institute (University of Cambridge). He was then awarded a Sir Henry Wellcome Postdoctoral Fellowship between The Institute of Cancer Research (ICR) (with Dr. Claus Jorgensen and Prof. Chris Marshall) and Massachusetts Institute of Technology (MIT) (with Prof. Doug Lauffenburger) to study how oncogenes signal across multiple cell types in cancer. Chris now leads the Cell-Communication Lab at UCL CI under a CRUK Career Development Fellowship (supported by the CRUK Werth Trust).
Keywords
Organoids, Barcoding, PTM Signalling
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