Multi-modal in situ profiling of human cells using the AVITI24 platform characterizes single-cell responses to acute stress
- 1SUNY Buffalo
Abstract
Cells begin to respond to stress within minutes, but how this early response depends on cell type, neighboring cells, or cell cycle phase has been difficult to determine, because bulk assays average across cells and synchronization can induce stress. We measured 338 transcripts, 48 proteins and phosphoproteins, and six cell paint stains in situ in 1.18 million single cells from four human cell lines exposed to H2O2, alone and in co-culture, and in A549 cells exposed to TNF-, cadmium, dithiothreitol or lipopolysaccharide. In A549 cells, five distinct stressors changed the same targets in the same direction within 15 min, differing mainly in magnitude, and modestly increased JNK phosphorylation. By 30 min of H2O2 treatment, immediate-early genes (FOS, JUN, DDIT3, DUSP1, NR4A1) were induced most strongly in HCT116 and A549 cells. These changes occurred across the whole cell population rather than in a responsive subset. Cell line-specific differences persisted in co-culture, and the response of MCF7 cells did not increase with the number of adjacent, strongly-responding A549 cells. Ordering cells along the cell cycle from the same measurements identified 106 cycle-associated transcripts, and the response varied more between cell lines than among cell cycle phases. Overall, the measured regulators respond to stress within 30 minutes through a largely shared program whose magnitude depends more on cell identity than on cell cycle state or neighboring cell type.
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BibTeX
@misc{lang2026multimodal,
title = {{Multi-modal in situ profiling of human cells using the AVITI24 platform characterizes single-cell responses to acute stress}},
author = {O. W. Lang and A. M. Gardner and C. M. Smugereski and A. M. Lyndaker and B. F. Pugh and W. Lai},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.28.762240},
url = {https://www.biorxiv.org/content/10.64898/2026.10.28.762240v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Multi-modal in situ profiling of human cells using the AVITI24 platform characterizes single-cell responses to acute stress
AU - O. W. Lang
AU - A. M. Gardner
AU - C. M. Smugereski
AU - A. M. Lyndaker
AU - B. F. Pugh
AU - W. Lai
PY - 2026
DA - 2026/10/28
PB - bioRxiv
DO - 10.64898/2026.10.28.762240
UR - https://www.biorxiv.org/content/10.64898/2026.10.28.762240v1
AB - Cells begin to respond to stress within minutes, but how this early response depends on cell type, neighboring cells, or cell cycle phase has been difficult to determine, because bulk assays average across cells and synchronization can induce stress. We measured 338 transcripts, 48 proteins and phosphoproteins, and six cell paint stains in situ in 1.18 million single cells from four human cell lines exposed to H2O2, alone and in co-culture, and in A549 cells exposed to TNF-, cadmium, dithiothreitol or lipopolysaccharide. In A549 cells, five distinct stressors changed the same targets in the same direction within 15 min, differing mainly in magnitude, and modestly increased JNK phosphorylation. By 30 min of H2O2 treatment, immediate-early genes (FOS, JUN, DDIT3, DUSP1, NR4A1) were induced most strongly in HCT116 and A549 cells. These changes occurred across the whole cell population rather than in a responsive subset. Cell line-specific differences persisted in co-culture, and the response of MCF7 cells did not increase with the number of adjacent, strongly-responding A549 cells. Ordering cells along the cell cycle from the same measurements identified 106 cycle-associated transcripts, and the response varied more between cell lines than among cell cycle phases. Overall, the measured regulators respond to stress within 30 minutes through a largely shared program whose magnitude depends more on cell identity than on cell cycle state or neighboring cell type.
ER -