Journal Article

Cell-type-specific signaling networks in heterocellular organoids

Li, Vivian S. W., Tape, Christopher J., Vlckova, Petra, Qin, Xiao, Sufi, Jahangir, Acton, Sophie E., Nitz, Mark, Kyriakidou, Pelagia

2020

Overview

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 (TOB is ) enables high-throughput comparison of signaling networks between organoid cultures. Cell-type-specific PTM analysis of colorectal cancer organoid cocultures reveals that shApc , Kras G12D and Trp53 R172H 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. Mass cytometry in combination with a thiol-reactive barcoding strategy allows analysis and comparison of cell-type-specific signaling networks in organoids.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 631/114/2391 / 631/61/475 / 631/67/327 / 631/80/458/1733 / 631/80/86 / Animals / Bioinformatics / Biological Microscopy / Biological models / Biological Techniques / Biomedical and Life Sciences / Biomedical Engineering/Biotechnology / Biomimetics / Cell Differentiation / Cell research / Chemical bonds / Coculture Techniques - methods / Colorectal cancer / Colorectal Neoplasms - metabolism / Colorectal Neoplasms - pathology / Cytometry / Cytophotometry - methods / Enterocytes / Enterocytes - cytology / Enteroendocrine Cells - cytology / Female / Fibroblasts / Fibroblasts - cytology / Gene Expression Regulation / Goblet cells / Goblet Cells - cytology / Humans / Intestine / Intestine, Small - cytology / Life Sciences / Macrophages / Macrophages - cytology / Mice / Mice, Inbred C57BL / Multivariate analysis / Networks / Organ Culture Techniques / Organoids / Organoids - metabolism / Organs (Anatomy) / Paneth Cells - cytology / Physiological aspects / Post-translation / Post-translational modification / Proteomics / Signal Transduction / Signaling / Single-Cell Analysis - methods / Sulfhydryl Compounds - chemistry / Tissue culture / Tumor Suppressor Protein p53 - metabolism