Cytometry‐based single‐cell analysis of intact epithelial signaling reveals MAPK activation divergent from TNF‐α‐induced apoptosis in vivo
Year: 2015
Type: article
Abstract: 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-α) stimulation. Unsupervised and supervised analyses robustly selected signaling features that identify a unique subset of epithelial cells that are sensitized to TNF-α-induced 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 multi-parameter flow cytometry, for investigating normal and diseased cell states in a wide range of epithelial tissues.
Source: Molecular Systems Biology
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $7,990
Funders: Crohn's and Colitis Foundation of America, Vanderbilt University, Vanderbilt Institute for Clinical and Translational Research, Vanderbilt-Ingram Cancer Center, Crohn's and Colitis Foundation, National Institutes of Health, Vanderbilt Digestive Diseases Research Center, Vanderbilt University Medical Center, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, National Institute of Diabetes and Digestive and Kidney Diseases, National Center for Advancing Translational Sciences, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1002678292, G1226162915, G1764958080, G1806196891, G1841051149, G1971342940, G2268269956, G2635048579, G292747457, G3080744291, G3269002440, G3371778737, G344109072, G3715404078, G3977540851, G407372463, G424458604, G4419210040, G454225982, SPORE in GI Cancer, Initiative for Maximizing Student Development, G5249137464, G548677123, G5526428480, G6569741389, G6793048653, G6832992462, G7029265662, G725763919, G7642566612, The Vanderbilt Institute for Clinical and Translational Research (VICTR), G8679799969, G8953775245, G9000001200, G9000064184, G9000076948, G9000089795, G9000109521, G9000130510, G9000241173, G9000244934, G9000263534, G9000322594, Molecular and Cellular Basis for Digestive Diseases, G991609106
OpenAlex ID: W2180807405