7 research outputs found
Predictive Endocrine Testing in the 21st Century Using <i>in Vitro</i> Assays of Estrogen Receptor Signaling Responses
Thousands
of environmental chemicals are subject to regulatory review for their
potential to be endocrine disruptors (ED). <i>In vitro</i> high-throughput screening (HTS) assays have emerged as a potential
tool for prioritizing chemicals for ED-related whole-animal tests.
In this study, 1814 chemicals including pesticide active and inert
ingredients, industrial chemicals, food additives, and pharmaceuticals
were evaluated in a panel of 13 <i>in vitro</i> HTS assays.
The panel of <i>in vitro</i> assays interrogated multiple
end points related to estrogen receptor (ER) signaling, namely binding,
agonist, antagonist, and cell growth responses. The results from the <i>in vitro</i> assays were used to create an ER Interaction Score.
For 36 reference chemicals, an ER Interaction Score >0 showed 100%
sensitivity and 87.5% specificity for classifying potential ER activity.
The magnitude of the ER Interaction Score was significantly related
to the potency classification of the reference chemicals (<i>p</i> < 0.0001). ERα/ERβ selectivity was also
evaluated, but relatively few chemicals showed significant selectivity
for a specific isoform. When applied to a broader set of chemicals
with <i>in vivo</i> uterotrophic data, the ER Interaction
Scores showed 91% sensitivity and 65% specificity. Overall, this study
provides a novel method for combining <i>in vitro</i> concentration
response data from multiple assays and, when applied to a large set
of ER data, accurately predicted estrogenic responses and demonstrated
its utility for chemical prioritization
An “EAR” on Environmental Surveillance and Monitoring: A Case Study on the Use of Exposure–Activity Ratios (EARs) to Prioritize Sites, Chemicals, and Bioactivities of Concern in Great Lakes Waters
Current
environmental monitoring approaches focus primarily on
chemical occurrence. However, based on concentration alone, it can
be difficult to identify which compounds may be of toxicological concern
and should be prioritized for further monitoring, in-depth testing,
or management. This can be problematic because toxicological characterization
is lacking for many emerging contaminants. New sources of high-throughput
screening (HTS) data, such as the ToxCast database, which contains
information for over 9000 compounds screened through up to 1100 bioassays,
are now available. Integrated analysis of chemical occurrence data
with HTS data offers new opportunities to prioritize chemicals, sites,
or biological effects for further investigation based on concentrations
detected in the environment linked to relative potencies in pathway-based
bioassays. As a case study, chemical occurrence data from a 2012 study
in the Great Lakes Basin along with the ToxCast effects database were
used to calculate exposure–activity ratios (EARs) as a prioritization
tool. Technical considerations of data processing and use of the ToxCast
database are presented and discussed. EAR prioritization identified
multiple sites, biological pathways, and chemicals that warrant further
investigation. Prioritized bioactivities from the EAR analysis were
linked to discrete adverse outcome pathways to identify potential
adverse outcomes and biomarkers for use in subsequent monitoring efforts
Real-Time Growth Kinetics Measuring Hormone Mimicry for ToxCast Chemicals in T‑47D Human Ductal Carcinoma Cells
High-throughput
screening (HTS) assays capable of profiling thousands of environmentally
relevant chemicals for <i>in vitro</i> biological activity
provide useful information on the potential for disrupting endocrine
pathways. Disruption of the estrogen signaling pathway has been implicated
in a variety of adverse health effects including impaired development,
reproduction, and carcinogenesis. The estrogen-responsive human mammary
ductal carcinoma cell line T-47D was exposed to 1815 ToxCast chemicals
comprising pesticides, industrial chemicals, pharmaceuticals, personal
care products, cosmetics, food ingredients, and other chemicals with
known or suspected human exposure potential. Cell growth kinetics
were evaluated using real-time cell electronic sensing. T-47D cells
were exposed to eight concentrations (0.006–100 μM),
and measurements of cellular impedance were repeatedly recorded for
105 h. Chemical effects were evaluated based on potency (concentration
at which response occurs) and efficacy (extent of response). A linear
growth response was observed in response to prototypical estrogen
receptor agonists (17β-estradiol, genistein, bisphenol A, nonylphenol,
and 4-<i>tert</i>-octylphenol). Several compounds, including
bisphenol A and genistein, induced cell growth comparable in efficacy
to that of 17β-estradiol, but with decreased potency. Progestins,
androgens, and corticosteroids invoked a biphasic growth response
indicative of changes in cell number or cell morphology. Results from
this cell growth assay were compared with results from additional
estrogen receptor (ER) binding and transactivation assays. Chemicals
detected as active in both the cell growth and ER receptor binding
assays demonstrated potencies highly correlated with two ER transactivation
assays (<i>r</i> = 0.72; <i>r</i> = 0.70). While
ER binding assays detected chemicals that were highly potent or efficacious
in the T-47D cell growth and transactivation assays, the binding assays
lacked sensitivity in detecting weakly active compounds. In conclusion,
this cell-based assay rapidly detects chemical effects on T-47D growth
and shows potential, in combination with other HTS assays, to detect
environmentally relevant chemicals with potential estrogenic activity
Real-Time Growth Kinetics Measuring Hormone Mimicry for ToxCast Chemicals in T‑47D Human Ductal Carcinoma Cells
High-throughput
screening (HTS) assays capable of profiling thousands of environmentally
relevant chemicals for <i>in vitro</i> biological activity
provide useful information on the potential for disrupting endocrine
pathways. Disruption of the estrogen signaling pathway has been implicated
in a variety of adverse health effects including impaired development,
reproduction, and carcinogenesis. The estrogen-responsive human mammary
ductal carcinoma cell line T-47D was exposed to 1815 ToxCast chemicals
comprising pesticides, industrial chemicals, pharmaceuticals, personal
care products, cosmetics, food ingredients, and other chemicals with
known or suspected human exposure potential. Cell growth kinetics
were evaluated using real-time cell electronic sensing. T-47D cells
were exposed to eight concentrations (0.006–100 μM),
and measurements of cellular impedance were repeatedly recorded for
105 h. Chemical effects were evaluated based on potency (concentration
at which response occurs) and efficacy (extent of response). A linear
growth response was observed in response to prototypical estrogen
receptor agonists (17β-estradiol, genistein, bisphenol A, nonylphenol,
and 4-<i>tert</i>-octylphenol). Several compounds, including
bisphenol A and genistein, induced cell growth comparable in efficacy
to that of 17β-estradiol, but with decreased potency. Progestins,
androgens, and corticosteroids invoked a biphasic growth response
indicative of changes in cell number or cell morphology. Results from
this cell growth assay were compared with results from additional
estrogen receptor (ER) binding and transactivation assays. Chemicals
detected as active in both the cell growth and ER receptor binding
assays demonstrated potencies highly correlated with two ER transactivation
assays (<i>r</i> = 0.72; <i>r</i> = 0.70). While
ER binding assays detected chemicals that were highly potent or efficacious
in the T-47D cell growth and transactivation assays, the binding assays
lacked sensitivity in detecting weakly active compounds. In conclusion,
this cell-based assay rapidly detects chemical effects on T-47D growth
and shows potential, in combination with other HTS assays, to detect
environmentally relevant chemicals with potential estrogenic activity
Real-Time Growth Kinetics Measuring Hormone Mimicry for ToxCast Chemicals in T‑47D Human Ductal Carcinoma Cells
High-throughput
screening (HTS) assays capable of profiling thousands of environmentally
relevant chemicals for <i>in vitro</i> biological activity
provide useful information on the potential for disrupting endocrine
pathways. Disruption of the estrogen signaling pathway has been implicated
in a variety of adverse health effects including impaired development,
reproduction, and carcinogenesis. The estrogen-responsive human mammary
ductal carcinoma cell line T-47D was exposed to 1815 ToxCast chemicals
comprising pesticides, industrial chemicals, pharmaceuticals, personal
care products, cosmetics, food ingredients, and other chemicals with
known or suspected human exposure potential. Cell growth kinetics
were evaluated using real-time cell electronic sensing. T-47D cells
were exposed to eight concentrations (0.006–100 μM),
and measurements of cellular impedance were repeatedly recorded for
105 h. Chemical effects were evaluated based on potency (concentration
at which response occurs) and efficacy (extent of response). A linear
growth response was observed in response to prototypical estrogen
receptor agonists (17β-estradiol, genistein, bisphenol A, nonylphenol,
and 4-<i>tert</i>-octylphenol). Several compounds, including
bisphenol A and genistein, induced cell growth comparable in efficacy
to that of 17β-estradiol, but with decreased potency. Progestins,
androgens, and corticosteroids invoked a biphasic growth response
indicative of changes in cell number or cell morphology. Results from
this cell growth assay were compared with results from additional
estrogen receptor (ER) binding and transactivation assays. Chemicals
detected as active in both the cell growth and ER receptor binding
assays demonstrated potencies highly correlated with two ER transactivation
assays (<i>r</i> = 0.72; <i>r</i> = 0.70). While
ER binding assays detected chemicals that were highly potent or efficacious
in the T-47D cell growth and transactivation assays, the binding assays
lacked sensitivity in detecting weakly active compounds. In conclusion,
this cell-based assay rapidly detects chemical effects on T-47D growth
and shows potential, in combination with other HTS assays, to detect
environmentally relevant chemicals with potential estrogenic activity
Real-Time Growth Kinetics Measuring Hormone Mimicry for ToxCast Chemicals in T‑47D Human Ductal Carcinoma Cells
High-throughput
screening (HTS) assays capable of profiling thousands of environmentally
relevant chemicals for <i>in vitro</i> biological activity
provide useful information on the potential for disrupting endocrine
pathways. Disruption of the estrogen signaling pathway has been implicated
in a variety of adverse health effects including impaired development,
reproduction, and carcinogenesis. The estrogen-responsive human mammary
ductal carcinoma cell line T-47D was exposed to 1815 ToxCast chemicals
comprising pesticides, industrial chemicals, pharmaceuticals, personal
care products, cosmetics, food ingredients, and other chemicals with
known or suspected human exposure potential. Cell growth kinetics
were evaluated using real-time cell electronic sensing. T-47D cells
were exposed to eight concentrations (0.006–100 μM),
and measurements of cellular impedance were repeatedly recorded for
105 h. Chemical effects were evaluated based on potency (concentration
at which response occurs) and efficacy (extent of response). A linear
growth response was observed in response to prototypical estrogen
receptor agonists (17β-estradiol, genistein, bisphenol A, nonylphenol,
and 4-<i>tert</i>-octylphenol). Several compounds, including
bisphenol A and genistein, induced cell growth comparable in efficacy
to that of 17β-estradiol, but with decreased potency. Progestins,
androgens, and corticosteroids invoked a biphasic growth response
indicative of changes in cell number or cell morphology. Results from
this cell growth assay were compared with results from additional
estrogen receptor (ER) binding and transactivation assays. Chemicals
detected as active in both the cell growth and ER receptor binding
assays demonstrated potencies highly correlated with two ER transactivation
assays (<i>r</i> = 0.72; <i>r</i> = 0.70). While
ER binding assays detected chemicals that were highly potent or efficacious
in the T-47D cell growth and transactivation assays, the binding assays
lacked sensitivity in detecting weakly active compounds. In conclusion,
this cell-based assay rapidly detects chemical effects on T-47D growth
and shows potential, in combination with other HTS assays, to detect
environmentally relevant chemicals with potential estrogenic activity
Identification of Chemical Features Linked to Thyroperoxidase Inhibition
Presented at the Annual Society of Toxicology meetin