In vivo Imaging Analysis of Steroid-Nuclear Receptor Function

Division Of Basic Sciences - Nci

Investigators

Linked publications, trials & patents

Abstract

1. With fusion chimeras between GR and other nuclear receptors that manifest ligand dependent cytoplasmic/nuclear translocation, we developed assays using the chimeric receptor to detect hormone levels in water supplies and other environmental sources. The approach has now been successfully developed for the glucocorticoid receptor (GR), the retinoic acid receptor (RAR), the estrogen receptor (ER), the aryl hydrocarbon receptor (AhR), the progesterone receptor (PR), and the thyroid hormone receptor (ThR). We developed cell lines which express fluorescently tagged versions of the receptor chimeras. We have utilized this fluorescent protein fusion technology to show that U.S. water supplies are heavily contaminated with chemicals that have multiple hormone activities. In addition to the previous US patent, a Japanese patent was granted covering licensing of this technology. We have collaborated with multiple intramural and extramural investigators to track hormone contamination in U.S. waterways and other environments, and to explore the implication of these findings for human health. 2. Water sources are frequently contaminated with natural and anthropogenic substances having known or suspected endocrine disrupting activities; however, these activities are not routinely measured and monitored. Phenotypic bioassays are a promising new approach for detection and quantitation of endocrine disrupting chemicals (EDCs). We developed cell lines expressing fluorescent chimeric constructs capable of detecting environmental contaminants which interact with multiple nuclear receptors. Using these assays, we tested water samples collected in the summers of 2016 through 2019 from two major Virginia rivers. Samples were concentrated 200X and screened for contaminants interacting with the androgen (AR), glucocorticoid (GR), aryl hydrocarbon (AhR) and thyroid receptors. Among 45 tested sites, over 70% had AR activity and 60% had AhR activity. Many sites were also positive for GR and TRB activation (22% and 42%, respectively). Multiple sites were positive for more than one type of contaminants, indicating presence of complex mixtures. These activities may negatively impact river ecosystems and consequently human health.