Tox21 Projects

Cross-Partner Projects 

To accommodate the expanded strategic direction and focus of the Tox21 collaboration, a central functional group called the cross-partner project was created in 2018. 

BioPlanet 

Through this project, Tox21 researchers are working to annotate pathways by source, species, biological function/process, disease/toxicity relevance and availability of probing assays using data from the PubChem and ToxCast databases. The BioPlanet database currently hosts approximately 1,700 unique human pathways that have been thoroughly curated. A public web browser allows easy pathway visualization and analysis. Continued development of this resource will include compound activity data, sequence data, gene/protein expression data and pathway data from non-human species, in addition to organizing assays according to pathways/diseases/toxicity end points. This integrated platform will push forward systematic analysis and modeling of toxicity responses.

Assays

Tox21 researchers work on a variety of different assays: 

• Phenotypic readouts
  • Cytotoxicity assays (i.e., cell viability [measures ATP], kinetic measurement of cytotoxicity and viability in cells)
  • Apoptosis assays (i.e., caspase assays [measures activity of caspase 3/7*, 8, 9])
  • Membrane integrity assays (i.e., LDH and protease release)
  • Mitochondrial toxicity assays (i.e., mitochondrial membrane potential assay*)
  • Gene toxicity assays
    • Differential cytotoxicity (DNA damage repair gene–deficient cell lines, DT40 and mouse cell lines)
    • ATADS (ELG1)*
    • Micronucleus assay
    • Phospholipidosis assays
    • Genetic variation (more than 1,000 HapMap lines)
• Pathway-specific toxicological mechanisms
  • Pathway assays using reporters (e.g., luciferase, β-lactamase) measuring hypoxia, ER stress, NF-κB*, p53*, ARE/Nrf2*, HSE, CREB, AP-1, STAT, NFAT, HIF-1α*, ESRE*, real time cytotoxicity and viability*
  • Developmental pathway assays (retinol signaling*, Hedgehog/Gli* and SBE/SMAD*)
• Target-specific toxicological mechanisms
  • Nuclear receptor assays, including AR*, AhR*, ERα/β*, FXR*, GR*, LXR, PPARα*, PPARδ*, PPARγ*, PR*, PXR*, RXR*, TRβ*, VDR*, RORγ, CAR*, ERR*
  • G-protein-coupled receptor/calcium signaling assays, including TRHR*, TSHR*, GnRHR*, CHRM1*, HTR2A*, and KISS1R
  • G-protein-coupled receptor/cAMP signaling assays, including DRD2*, ADRB2*, and ADRB1*
  • Enzyme activity assays, including AChE*
  • hERG potassium channel assay*
  • Cytokine assays including IL-8 and TNF-α
  • Cytochrome P450 assay including CYP 1A2*, CYP 2C9*, CYP 2C19*, CYP 2D6*, CYP 3A4*, and CYP26A1*

* Assays screened against the Tox21 10K library

Neuronal Toxicity

Tox21 researchers are developing cellular models and methods to assess neurodegenerative and neurodevelopmental toxicities. Stem cell–derived, LUHMES-preserved and 3-D cultured neuronal models are included. Screening methods include neurite growth measurements and toxicant-chemical interaction “Toxmatrix” screening.

High Throughput Genomics

NCATS is developing methodologies to improve the throughput of tracking RNA changes of cellular models treated with various environmental, pharmaceutical and food-based compounds in the hopes of identifying signatures that affects our health.