Abstract
The sodium-iodide symporter (NIS) mediates the uptake of iodide into the thyroid. Inhibition of NIS function by xenobiotics has been demonstrated to suppress circulating thyroid hormones and perturb related physiological functions. Until recently, few environmental chemicals had been screened for NIS inhibition activity. We previously screened over 1000 chemicals from the ToxCast Phase II (ph1v2 and ph2) libraries using an in vitro radioactive iodide uptake (RAIU) with the hNIS-HEK293T cell line to identify NIS inhibitors. Here, we broaden the chemical space by expanding screening to include the ToxCast e1k library (804 unique chemicals) with initial screening for RAIU at 1 × 10–4 M. Then 209 chemicals demonstrating > 20% RAIU inhibition were further tested in multiple-concentration, parallel RAIU and cell viability assays. This identified 55 chemicals as active, noncytotoxic RAIU inhibitors. Further cytotoxicity-adjusted potency scoring (with NaClO4 having a reference score of 200) revealed five chemicals with moderate to strong RAIU inhibition (scored > 100). These data were combined with our previous PhII screening data to produce binary hit-calls for ~ 1800 unique chemicals (PhII + e1k) with and without cytotoxicity filtering. Results were analyzed with a ToxPrint chemotype-enrichment workflow to identify substructural features significantly enriched in the NIS inhibition hit-call space. We assessed the applicability of enriched PhII chemotypes to prospectively predict NIS inhibition in the e1k dataset. Chemotype enrichments derived for the combined ~ 1800 dataset also identified additional enriched features, as well as chemotypes affiliated with cytotoxicity. These enriched chemotypes provide important new information that can support future data interpretation, structure–activity relationship, chemical use, and regulation.
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Abbreviations
- AOP:
-
Adverse outcome pathway
- 3bMAD:
-
Three times the baseline median absolute deviation
- CV:
-
Coefficient of variation
- CPM:
-
Counts per minute
- CT:
-
Chemotype
- CTEW:
-
Chemotype enrichment workflow
- DCNQ:
-
2,3-Dichloro-1,4-napthoquinone
- DMEM:
-
Dulbecco’s Modified Eagle Medium
- DMSO:
-
Dimethyl sulfoxide
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- EDSP21:
-
Endocrine Disruptor Screening Program in the twenty-first century
- FDA-GRAS:
-
U.S. Food & Drug Administration’s Generally Regarded as Safe program
- HEK:
-
Human embryonic kidney
- HTS:
-
High-throughput screening
- NIS:
-
Sodium iodide symporter
- PFOS:
-
Perfluorooctanesulfonic acid
- PFOA:
-
Perfluorooctanoic acid
- IRIS:
-
Integrated Risk Information System
- RAIU:
-
Radioactive iodide uptake
- RLU:
-
Relative light unit
- SD:
-
Standard deviation
- T3 :
-
Triiodothyronine
- ToxCast:
-
Toxicity Forecaster
- T4 :
-
Thyroxine
- TH:
-
Thyroid hormone
- TAA:
-
Toxicity-adjusted area
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Acknowledgements
This research was funded by the Office of Research and Development, U.S. EPA, Washington, DC. The authors thank U.S. EPA scientists Jennifer Olker for reviewing the early draft, Christopher Grulke and Antony Williams for computational chemistry expertise and support, and Chihae Yang and colleagues at Molecular Networks-Altamira (MN-AM) for inspirational development of the ToxPrints and Chemotyper used in this study.
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Wang, J., Richard, A.M., Murr, A.S. et al. Expanded high-throughput screening and chemotype-enrichment analysis of the phase II: e1k ToxCast library for human sodium-iodide symporter (NIS) inhibition. Arch Toxicol 95, 1723–1737 (2021). https://doi.org/10.1007/s00204-021-03006-2
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DOI: https://doi.org/10.1007/s00204-021-03006-2