Profiling of Environmental Chemicals on Nuclear Receptors
Nuclear receptors (NRs) play critical roles in development, metabolic homeostasis, and reproduction. Inappropriate activation of the nuclear receptors by environmental chemicals will lead to a broad spectrum of adverse effects. To assess environmental chemicals on endocrine disruption, the Tox21 collaboration has selected appropriate NR assays in combing with quantitative high-throughput screening (qHTS). As a proof-of-concept study, the Tox21 pilot phase collection of approximately 3,000 environmental chemicals was profiled against a panel of ten human nuclear receptors in qHTS format. We have evaluated these data from a chemical genomics point of view, in addition to assessing data reproducibility as a measure of data quality and applicability for down-stream analyses.
Nuclear receptors interacting with genes
Example structure classes with consistent nuclear
receptor activity patterns or signatures
Tox21 Consortium Members
National Institute of Environmental Health Sciences/National Toxicology Program
U.S. Environmental Protection Agency
NIH Chemical Genomics Center, NCATS
U.S. Food and Drug Administration
Key Investigators
National Center for Advancing Translational Sciences
Menghang Xia, Ph.D.
Ruili Huang, Ph.D.
David Gerhold, Ph.D.
Anton Simeonov, Ph.D.
Christopher P. Austin, M.D.
Public Health Impact
Data production from Tox21 is now moving into its exponential growth phase. Over the next several years, the interdisciplinary Tox21 collaboration will continue to innovate in assay biology, screening and computation to usher in a new era of efficient, mechanistic and predictive chemical toxicology.
Publications
Huang R, Xia M, Cho MH, et al. Chemical genomics profiling of environmental chemical modulation of human nuclear receptors. Environ Health Perspect, 2011;119:1142-1148.
Martin MT, Knudsen TB, Reif DM, et al. Predictive model of rat reproductive toxicity from ToxCast high throughput screening. Biol Reprod, 2011;85:327-339.
Judson RS, Martin MT, Reif DM, et al. Analysis of eight oil spill dispersants using rapid, in vitro testes for endocrine and other biological activity. Environ Sci Technol, 2011;44:5979-5985.
Shukla SJ, Sakamuru S, Huang R, et al. Identification of clinically utilized drugs that activate pregnane X receptors. Drug Metab Dispos, 2011;39:151-159.
Shukla SJ, Huang R, Austin CP, Xia M. The future of toxicity testing: a focus on in vitro methods using a quantitative high throughput screening platform. Drug Discov Today, 2011;15:997-1007.
Project Details
Tox21 assays currently available at NCGC
| Assay | Assay Endpointa | Cell Type | Assay Readout |
|---|---|---|---|
| Cell viability Apoptosis | Intracellular ATP Caspase-3/7 | Hek293; Jurkat;HepG2; Sh-SY5Y; SK-N-SH; BJ; HUV-EC-C; MRC-5; mesangial; kidney proximal tubules; N2a; H-4-II-E;NIH3T3 | Luminescence |
| Membrane integrity | LDH release Protease release | HEK293; mesangial | Flourescence Luminescence |
| Mitochondrial toxicity | Membrane potential | HepG2 | Flourescence |
| DNA damage | Micronucleus | CHO | Flourescence |
| Cytokine | IL-8; TNF-α | THP-1 | Homogenous time-resolved flourescence |
| Nuclear receptor | AR; ERα; FXR; PPARδ; PPARγ; RXR; TRβ; VDR; GR hPXR; AhR; rPXR | Hek293 HeLa HepG2 | β-Lactamase reporter Luciferase reporter |
| Toxicity pathway | AP-1; HIF-1α SIE; NFKB; HSR; ESRE ARE/Nrf2 CREB p53 ARE/Nrf2; HSR; ESRE | ME-180 HeLa HepG2 Hek293, CHO HCT-116 HepG2 | β-Lactamase reporter Luciferase reporter |
| hERG channel | Thallium influx | U-2OS | Flourescence |
a AhR, aryl hydrocarbon receptor; AP-1, activator protein-1; AR, androgen receptor; ARE/Nrf2, antioxidant response element/NF-E2 related factor 2; CREB, cAMP response element binding; ERa, estrogen receptor a; ESRE, endoplasmic reticulum stress response element; FXR, farnesoid X receptor; GR, glucocorticoid receptor; HIF-1a, hypoxia-inducible factor-1a; hPXR, human pregnane X receptor; HSE, heat shock response element; IL-8, interleukin-8; LDH, lactate dehydrogenase; NFkB, nuclear factor kappa B; PPARd, peroxisome proliferator-activated receptor d; PPARg, peroxisome proliferator-activated receptor g; rPXR, rat pregnane X receptor; RXR, retinoid X receptor; SIE, sis-inducible element; TNFa, tumor necrosis factor a; TRb, thyroid hormone receptor b; VDR, vitamin D receptor.
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