Open AccessCytometry‐based single‐cell analysis of intact epithelial signaling reveals MAPK activation divergent from TNF ‐α‐induced apoptosis in vivo
Author(s) -
Simmons Alan J,
Banerjee Amrita,
McKinley Eliot T,
Scurrah Cherie’ R,
Herring Charles A,
Gewin Leslie S,
Masuzaki Ryota,
Karp Seth J,
Franklin Jeffrey L,
Gerdes Michael J,
Irish Jonathan M,
Coffey Robert J,
Lau Ken S
Publication year - 2016
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20167270
Subject(s) - irish , philosophy , linguistics
Understanding heterogeneous cellular behaviors in a complex tissue requires the evaluation of signaling networks at single-cell resolution. However, probing signaling in epithelial tissues using cytometry-based single-cell analysis has been confounded by the necessity of single-cell dissociation, where disrupting cell-to-cell connections inherently perturbs native cell signaling states. Here, we demonstrate a novel strategy (Disaggregation for Intracellular Signaling in Single Epithelial Cells from Tissue—DISSECT) that preserves native signaling for Cytometry Time-of-Flight (CyTOF) and fluorescent flow cytometry applications. A 21-plex CyTOF analysis encompassing core signaling and cell-identity markers was performed on the small intestinal epithelium after systemic tumor necrosis factor-alpha (TNF-a) stimulation. Unsupervised and supervised analyses robustly selected signaling features that identify a unique subset of epithelial cells that are sensitized to TNF-ainduced apoptosis in the seemingly homogeneous enterocyte population. Specifically, p-ERK and apoptosis are divergently regulated in neighboring enterocytes within the epithelium, suggesting a mechanism of contact-dependent survival. Our novel single-cell approach can broadly be applied, using both CyTOF and multiparameter flow cytometry, for investigating normal and diseased cell states in a wide range of epithelial tissues.