Tox21 Research Phases
Data from all phases of Tox21, along with full chemical characterization and assay protocol details, are being deposited into publicly available databases, which are accessible through the Tox21 Toolbox.
Tox21 Research - Phase I
In the first phase of Tox21 research (2005 – 2010), the partners utilized the high throughput screening (HTS) technologies of the National Human Genome Research Institute's National Chemical Genomics Center (NCGC, now a part of the National Center for Advancing Translational Sciences, NCATS) and performed quantitative HTS (qHTS) testing (see box). NCGC screened approximately 2800 compounds in more than 75 qHTS assays. Concurrently, the Environmental Protection Agency’s National Center for Computational Toxicology screened 309 unique compounds across more than 500 biochemical- and cell-based assays through its ToxCast™ program. These compounds were also tested in NTP's "WormTox" laboratory, where toxicity screens used the roundworm, Caenorhabditis elegans.
Tox21 Research - Phase II
In Phase II of Tox21 research (2011 – 2016), the partners are testing a chemical library, expanded from Phase I to greater than 10,000 compounds, at NCATS using a HTS robotics system sponsored by NTP. More than 200 databases of chemicals and drugs in the United States and abroad were analyzed to select the compounds for testing. This chemical library includes industrial and consumer products, food additives, drugs, and mixtures. Tox21 partners are establishing a full spectrum of cell-based assays to further define and characterize activities identified in the initial qHTS tests. The results will provide information to help evaluate biological processes that may lead to adverse health effects in humans.
Tox21 Research - Phase III
Phase III of Tox21 research (2014 – present) aims to better represent human health and disease by increasing the biological diversity of human cell lines used in the HTS assays. Phase III plans to include three-dimensional organoid (or organ-like) model cultures and simple model organisms to address chemical metabolism and assess the effects of chemicals in more complex biological systems. Phase III research also adds medium to high throughput transcriptomics assays to test for effects of chemicals on gene expression. These Phase III screening assays will look at multiple endpoints and processes in order to better identify chemicals that may cause adverse human health effects.
During Phase III, Tox21 partners have identified sets of genes in human cells and tissues that respond to toxic chemicals (i.e., toxicogenomics - see box). The resulting human S1500+ gene set comprises approximately 2750 genes that will be used to evaluate gene expression changes from chemical exposures. Similar gene sets are being developed for screening cells and tissues from rats, mice, and zebrafish. The human S1500+ gene set should provide transcriptomic information on cellular responses that are independent of cell type or species, and also those that are specific by organ and/or cell type.