Impaired coordination between signaling pathways is revealed in human colorectal cancer using single-cell mass cytometry of archival tissue blocks
Year: 2016
Type: article
Abstract: Cellular heterogeneity poses a substantial challenge to understanding tissue-level phenotypes and confounds conventional bulk analyses. To analyze signaling at the single-cell level in human tissues, we applied mass cytometry using cytometry time of flight to formalin-fixed, paraffin-embedded (FFPE) normal and diseased intestinal specimens. This technique, called FFPE-DISSECT (disaggregation for intracellular signaling in single epithelial cells from tissue), is a single-cell approach to characterizing signaling states in embedded tissue samples. We applied FFPE-DISSECT coupled to mass cytometry and found differential signaling by tumor necrosis factor-α in intestinal enterocytes, goblet cells, and enteroendocrine cells, implicating the downstream RAS-RAF-MEK pathway in determining goblet cell identity. Application of this technique and computational analyses to human colon specimens confirmed the reduced differentiation in colorectal cancer (CRC) compared to normal colon and revealed increased intratissue and intertissue heterogeneity in CRC with quantitative changes in the regulation of signaling pathways. Specifically, coregulation of the kinases p38 and ERK, the translation regulator 4EBP1, and the transcription factor CREB in proliferating normal colon cells was lost in CRC. Our data suggest that this single-cell approach, applied in conjunction with genomic annotation, enables the rapid and detailed characterization of cellular heterogeneity from clinical repositories of embedded human tissues. This technique can be used to derive cellular landscapes from archived patient samples (beyond CRC) and as a high-resolution tool for disease characterization and subtyping.
Source: Science Signaling
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Funders: Vanderbilt Institute for Clinical and Translational Research, Crohn's and Colitis Foundation, National Institutes of Health, 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: G1113670494, G1331061530, G1405049813, G1806196891, G1971342940, G2284163681, Validation of a single cell multiplexed in-situ biomarker analysis platform for p, G292747457, G2933723276, G326042038, G3364599783, G3371778737, G3715404078, G3977540851, G407372463, SPORE in GI Cancer, Initiative for Maximizing Student Development, G548677123, Training Program in Stem Cell and Regenerative Developmental Biology, G725763919, The Vanderbilt Institute for Clinical and Translational Research (VICTR), G8679799969, Molecular and Cellular Basis for Digestive Diseases, G991609106
OpenAlex ID: W2531731128