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Cited by 1Per- and polyfluoroalkyl substances researchToxic Organic Pollutants ImpactCarcinogens and Genotoxicity Assessment

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Identification of functional genetic components modulating toxicity response to PFOS using genome-wide CRISPR screens in HepG2/C3A cells

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Kim, Chanhee, Tagmount, Abderrahmane, Zhu, Zhaohan, Wilson, Frances, Li, Danmeng, Ostrov, David A., Brad Barbazuk, William, Bacher, Rhonda, Vulpe, Chris D.·2026·Archives of Toxicology
DOI 10.1007/s00204-025-04294-8PMC PMC12724516

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Abstract

Perfluorooctane sulfonate (PFOS) poses significant health and environmental risks due to its persistence and widespread use and has been linked to various adverse outcomes, such as liver toxicity. Although the molecular responses and toxicity effects of PFOS exposure have been extensively studied, considerable uncertainty remains regarding the causal mechanisms leading to PFOS-associated adverse effects. To help bridge this gap, we conducted CRISPR screens in HepG2/C3A human liver cells exposed to IC 25 (170 µM) of PFOS to identify genes and pathways influencing PFOS-induced cytotoxicity. Using a genome-wide CRISPR knockout library targeting 18,819 genes, we identified 340 candidate genes that modulate PFOS-induced cytotoxicity when genetically disrupted (189 gene disruptions increased sensitivity and 151 gene disruptions increased resistance). From these candidate genes, we individually disrupted two candidate genes, SLC6A9 which encodes the glycine transporter GlyT1, and CPSF2 , and confirmed increased resistance to PFOS exposure. Further, molecular docking analysis predicts that PFOS directly binds to GlyT1 and functional inhibition of GlyT1 also increases resistance to PFOS exposure. Gene-Disease outcome association analysis using the Comparative Toxicogenomics Database (CTD) indicated an enrichment of candidate genes associated with cancer-related and liver disease phenotypes. KEGG and STRING enrichment analyses found over representation of several biological pathways including DNA damage response and cell cycle. Lastly, cross-species conservation analysis using the top two validated gene targets found that their pathways were highly conserved in several environmentally relevant species. These findings provide new mechanistic and functional insights into PFOS-induced cytotoxicity, highlight potential molecular targets for toxicity mitigation, and establish a foundation for cross-species toxicogenomic modeling of PFOS health effects.

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Metadata

Authors
Kim, C., Tagmount, A., Zhu, Z., Wilson, F., Li, D., Ostrov, D. A., Brad Barbazuk, W., Bacher, R., & Vulpe, C. D.
Affiliations
University of Florida (US)
Corresponding author
Kim, Chanhee
ORCID
Venue
Archives of Toxicology
Publisher
Springer Science and Business Media LLC
Published
2026-01-27
Volume
100
Issue
4
Pages
1391-1410
ISSN
  • 0340-5761 (ISSN-L)
  • 1432-0738
DOI
10.1007/s00204-025-04294-8
PMC
PMC12724516
OpenAlex
W7125790313
Cited by
1
References
7
Citations by year
2026: 1
FWCI
1.90
Type
journal article
Language
en
Open access
Licence
Locations
  • Archives of Toxicology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
National Institute of Environmental Health Sciences (R01ES033625)
Topics
  • Physical Sciences › Environmental Science › Environmental Chemistry › Per- and polyfluoroalkyl substances research
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Toxic Organic Pollutants Impact
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
Keywords
PFOS, PFOS toxicity, hepatotoxicity, cytotoxicity, CRISPR knockout screen, SLC6A9, glycine transporter 1, glycine transporter, gene disruption, DNA damage response, cell cycle, cross-species conservation, toxicogenomics, liver cancer, perfluorinated compounds

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