Multi-omics study links autophagy and ER stress to PFOA toxicity in sperm precursor cells

Perfluorooctanoic acid (PFOA), a persistent “forever chemical” of the per- and polyfluoroalkyl substance family, has been increasingly associated with male reproductive toxicity, but how its effects unfold inside germ cells has remained unclear.

The researchers found that PFOA significantly reduced cell viability and increased early apoptosis in a concentration-dependent manner. To uncover the molecular basis of this injury, the team performed integrated transcriptomic, proteomic and metabolomic analyses. These multi-omics profiles revealed coordinated alterations pointing to proteostasis disruption, redox imbalance, metabolic remodeling, endoplasmic reticulum (ER) stress and autophagy, suggesting that several stress systems respond together rather than in isolation when the cells encounter the chemical.

Mechanistically, the study reported that PFOA induced accumulation of p62, decreased LC3-II expression, and reduced both GFP-LC3 and mRFP-GFP-LC3 puncta, a pattern the authors describe as consistent with altered autophagic processing. To probe this further, the researchers used pharmacological tools: activation of autophagy with rapamycin or Torin1 modified autophagy-associated protein readouts, and rapamycin alleviated the PFOA-induced early apoptosis. In parallel, PFOA increased eIF2α-associated stress readouts, a hallmark of the integrated stress response, and the ER stress kinase inhibitor GSK2606414 partially attenuated early apoptosis in the exposed cells.

The team also added new oxidative-stress assays and an NAC intervention, which linked oxidative imbalance to several of the stress-associated readouts. Taken together, the findings implicate altered autophagic processing, ER and integrated-stress signaling, and oxidative imbalance in acute PFOA-induced injury of GC-1 cells. The authors are careful, however, to state the limits of the work: the study does not establish the causal order of these stress responses, and the results were obtained in an immortalized cell line rather than in primary spermatogonial stem cells, so the findings cannot yet be extended directly to the stem cells that sustain sperm production. As an open-access article shared early with a permanent DOI, the study offers a multi-omics framework for dissecting how persistent fluorinated chemicals stress germ cells, while leaving the question of which pathway drives the damage, and whether the same responses occur in the intact testis, for future research.

Subject of Research: Multi-omics analysis of autophagy and ER stress responses in PFOA-exposed GC-1 spermatogonial cells

Article Title: Integrated multi-omics and functional validation implicate autophagy and ER stress in PFOA-induced GC-1 cell toxicity

Article References: Duan, J., Sun, Y., Ye, W., Huang, S., Yuan, R., Zhang, B., Chen, X., Zhang, S., Lin, H., Liang, Y., Song, Y., & Xia, S. (2026). Integrated multi-omics and functional validation implicate autophagy and ER stress in PFOA-induced GC-1 cell toxicity. Cell Biology and Toxicology. https://doi.org/10.1007/s10565-026-10322-9

Image Credits: AI Generated

DOI: 10.1007/s10565-026-10322-9

Keywords: PFOA, PFAS, GC-1 spermatogonial cells, autophagy, ER stress, multi-omics, male reproductive toxicity, apoptosis, rapamycin, oxidative stress, integrated stress response, Cell Biology and Toxicology