High-throughput cytokine detection platform to identify potential neurotoxicants
Exposure to environmental chemicals, such as pesticides and heavy metals, may contribute to neurodegenerative disorders by inducing inflammation in nervous system tissues. EPA, NCATS, and NIEHS collaborated to develop in vitro models using human induced pluripotent stem cell-derived microglia to identify potential neurotoxicants (Yang et al. 2025). The assays measured cytokine secretion, which is a marker of inflammatory response. The cytokine response patterns in these cells closely resembled those under in vivo conditions, with significant increases in interleukin 6 and tumor necrosis factor-alpha levels. These initial results were used as the basis for development of a homogeneous, time-resolved fluorescence HTS assay platform in a 1,536-well plate format for screening of environmental chemicals. This assay was used to test nine environmental chemicals having evidence of cytokine-inducing potential in other in vitro assays. While all nine tested agents stimulated interleukin 6 and tumor necrosis factor-alpha production, three chemicals showed significant stimulation of both cytokines. This study establishes a reliable high-throughput platform for detecting inflammatory effects of environmental toxicants in a microglial cell assay, data from which may provide valuable insights into their neuroinflammatory potential and possible contributions to development of neurodegenerative disorders.