Cell-type-specific signaling networks in heterocellular organoids.
journal contributionposted on 07.09.2020, 11:04 by Xiao Qin, Jahangir Sufi, Petra Vlckova, Pelagia Kyriakidou, Sophie E Acton, Vivian SW Li, Mark Nitz, Christopher J Tape
Despite the widespread adoption of organoids as biomimetic tissue models, methods to comprehensively analyze cell-type-specific post-translational modification (PTM) signaling networks in organoids are absent. Here, we report multivariate single-cell analysis of such networks in organoids and organoid cocultures. Simultaneous analysis by mass cytometry of 28 PTMs in >1 million single cells derived from small intestinal organoids reveals cell-type- and cell-state-specific signaling networks in stem, Paneth, enteroendocrine, tuft and goblet cells, as well as enterocytes. Integrating single-cell PTM analysis with thiol-reactive organoid barcoding in situ (TOBis) enables high-throughput comparison of signaling networks between organoid cultures. Cell-type-specific PTM analysis of colorectal cancer organoid cocultures reveals that shApc, KrasG12D and Trp53R172H cell-autonomously mimic signaling 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 signaling analysis of healthy and cancerous organoids.
AnimalsBiomimeticsCell DifferentiationCoculture TechniquesColorectal NeoplasmsCytophotometryEnterocytesEnteroendocrine CellsFemaleFibroblastsGene Expression RegulationGoblet CellsHumansIntestine, SmallMacrophagesMiceMice, Inbred C57BLOrgan Culture TechniquesOrganoidsPaneth CellsSignal TransductionSingle-Cell AnalysisSulfhydryl CompoundsTumor Suppressor Protein p53Li, V FC00110506 Biological Sciences10 Technology11 Medical and Health SciencesDevelopmental Biology