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Updated: Sep 12, 2025

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors

Published on: February 7, 2018

Decoding Cellular Stress States for Toxicology Using Single-Cell Transcriptomics.

Imran Shah¹, David Gallegos¹, Brian Robinette¹

¹Center for Computational Toxicology and Exposure, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina, USA.

Biorxiv : the Preprint Server for Biology

|August 6, 2025

Summary

This summary is machine-generated.

Single-cell transcriptomics revealed diverse cellular stress responses to chemicals, identifying five distinct cell states from homeostasis to apoptosis. This approach aids in understanding toxicological pathways and adaptive cellular transitions.

Keywords:

HepaRG adaptive stress response cell states cell strate transition graph computational toxicology single-cell transcriptomics (SCTr)

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Last Updated: Sep 12, 2025

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors

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Published on: February 7, 2018

Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues

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Published on: January 10, 2019

Measurements of Physiological Stress Responses in C. Elegans

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Published on: May 21, 2020

Area of Science:

Toxicology
Cell Biology
Genomics

Background:

Understanding cellular responses to chemical exposure is crucial for toxicology.
Adaptive stress response pathways (SRPs) play a key role in cell survival and death.
Single-cell transcriptomic (SCTr) analysis offers high resolution for studying cellular heterogeneity.

Purpose of the Study:

To characterize adaptive stress response pathways (SRPs) in HepaRG cells using SCTr.
To investigate cellular responses to various chemical stressors at the single-cell level.
To identify distinct cellular phenotypic states and transitions during chemical exposure.

Main Methods:

Applied the TempO-LINC® platform for SCTr profiling of ~40,000 HepaRG cells.
Exposed cells to etoposide, brefeldin A, cycloheximide, rotenone, tBHQ, troglitazone, and tunicamycin.
Scored cells using SRP gene signatures and clustered phenotypes using a generalized Jaccard metric.

Main Results:

Identified five distinct phenotypic groups: homeostasis, adaptive responses, terminal outcomes, autophagy, and apoptosis.
Observed heterogeneous responses to chemicals, with specific pathways like UPR, OSR, HSR, and DDR activated.
Troglitazone upregulated lipid metabolism and HSR/UPR; Brefeldin A/Tunicamycin induced UPR; Rotenone triggered mitochondrial stress responses.

Conclusions:

SCTr is valuable for decoding cellular stress states and heterogeneity.
The study provides insights into transitions between cellular adaptation and toxicity.
This methodology can elucidate mechanisms underlying chemical-induced cell death.