10 research outputs found
Profiling 976 ToxCast Chemicals across 331 Enzymatic and Receptor Signaling Assays
Understanding potential health risks
is a significant challenge
due to the large numbers of diverse chemicals with poorly characterized
exposures and mechanisms of toxicities. The present study analyzes
976 chemicals (including failed pharmaceuticals, alternative plasticizers,
food additives, and pesticides) in Phases I and II of the U.S. EPA’s
ToxCast project across 331 cell-free enzymatic and ligand-binding
high-throughput screening (HTS) assays. Half-maximal activity concentrations
(AC50) were identified for 729 chemicals in 256 assays (7,135 chemical–assay
pairs). Some of the most commonly affected assays were CYPs (CYP2C9
and CYP2C19), transporters (mitochondrial TSPO, norepinephrine, and
dopaminergic), and GPCRs (aminergic). Heavy metals, surfactants, and
dithiocarbamate fungicides showed promiscuous but distinctly different
patterns of activity, whereas many of the pharmaceutical compounds
showed promiscuous activity across GPCRs. Literature analysis confirmed
>50% of the activities for the most potent chemical–assay
pairs
(54) but also revealed 10 missed interactions. Twenty-two chemicals
with known estrogenic activity were correctly identified for the majority
(77%), missing only the weaker interactions. In many cases, novel
findings for previously unreported chemical–target combinations
clustered with known chemical–target interactions. Results
from this large inventory of chemical–biological interactions
can inform read-across methods as well as link potential targets to
molecular initiating events in adverse outcome pathways for diverse
toxicities
Profiling 976 ToxCast Chemicals across 331 Enzymatic and Receptor Signaling Assays
Understanding potential health risks
is a significant challenge
due to the large numbers of diverse chemicals with poorly characterized
exposures and mechanisms of toxicities. The present study analyzes
976 chemicals (including failed pharmaceuticals, alternative plasticizers,
food additives, and pesticides) in Phases I and II of the U.S. EPA’s
ToxCast project across 331 cell-free enzymatic and ligand-binding
high-throughput screening (HTS) assays. Half-maximal activity concentrations
(AC50) were identified for 729 chemicals in 256 assays (7,135 chemical–assay
pairs). Some of the most commonly affected assays were CYPs (CYP2C9
and CYP2C19), transporters (mitochondrial TSPO, norepinephrine, and
dopaminergic), and GPCRs (aminergic). Heavy metals, surfactants, and
dithiocarbamate fungicides showed promiscuous but distinctly different
patterns of activity, whereas many of the pharmaceutical compounds
showed promiscuous activity across GPCRs. Literature analysis confirmed
>50% of the activities for the most potent chemical–assay
pairs
(54) but also revealed 10 missed interactions. Twenty-two chemicals
with known estrogenic activity were correctly identified for the majority
(77%), missing only the weaker interactions. In many cases, novel
findings for previously unreported chemical–target combinations
clustered with known chemical–target interactions. Results
from this large inventory of chemical–biological interactions
can inform read-across methods as well as link potential targets to
molecular initiating events in adverse outcome pathways for diverse
toxicities
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
Suspect Screening Analysis of Chemicals in Consumer Products
A two-dimensional gas chromatography-time-of-flight/mass
spectrometry
(GCĂ—GC-TOF/MS) suspect screening analysis method was used to
rapidly characterize chemicals in 100 consumer productsî—¸which
included formulations (e.g., shampoos, paints), articles (e.g., upholsteries,
shower curtains), and foods (cereals)î—¸and therefore supports
broader efforts to prioritize chemicals based on potential human health
risks. Analyses yielded 4270 unique chemical signatures across the
products, with 1602 signatures tentatively identified using the National
Institute of Standards and Technology 2008 spectral database. Chemical
standards confirmed the presence of 119 compounds. Of the 1602 tentatively
identified chemicals, 1404 were not present in a public database of
known consumer product chemicals. Reported data and model predictions
of chemical functional use were applied to evaluate the tentative
chemical identifications. Estimated chemical concentrations were compared
to manufacturer-reported values and other measured data. Chemical
presence and concentration data can now be used to improve estimates
of chemical exposure, and refine estimates of risk posed to human
health and the environment
Suspect Screening Analysis of Chemicals in Consumer Products
A two-dimensional gas chromatography-time-of-flight/mass
spectrometry
(GCĂ—GC-TOF/MS) suspect screening analysis method was used to
rapidly characterize chemicals in 100 consumer productsî—¸which
included formulations (e.g., shampoos, paints), articles (e.g., upholsteries,
shower curtains), and foods (cereals)î—¸and therefore supports
broader efforts to prioritize chemicals based on potential human health
risks. Analyses yielded 4270 unique chemical signatures across the
products, with 1602 signatures tentatively identified using the National
Institute of Standards and Technology 2008 spectral database. Chemical
standards confirmed the presence of 119 compounds. Of the 1602 tentatively
identified chemicals, 1404 were not present in a public database of
known consumer product chemicals. Reported data and model predictions
of chemical functional use were applied to evaluate the tentative
chemical identifications. Estimated chemical concentrations were compared
to manufacturer-reported values and other measured data. Chemical
presence and concentration data can now be used to improve estimates
of chemical exposure, and refine estimates of risk posed to human
health and the environment
Measuring Physicochemical Properties to Inform the Scope of Existing QSAR/QSPR Models
Presented at the Annual Society of Toxicology meetin
ToxCast Chemical Landscape: Paving the Road to 21st Century Toxicology
The
U.S. Environmental Protection Agency’s (EPA) ToxCast
program is testing a large library of Agency-relevant chemicals using <i>in vitro</i> high-throughput screening (HTS) approaches to support
the development of improved toxicity prediction models. Launched in
2007, Phase I of the program screened 310 chemicals, mostly pesticides,
across hundreds of ToxCast assay end points. In Phase II, the ToxCast
library was expanded to 1878 chemicals, culminating in the public
release of screening data at the end of 2013. Subsequent expansion
in Phase III has resulted in more than 3800 chemicals actively undergoing
ToxCast screening, 96% of which are also being screened in the multi-Agency
Tox21 project. The chemical library unpinning these efforts plays
a central role in defining the scope and potential application of
ToxCast HTS results. The history of the phased construction of EPA’s
ToxCast library is reviewed, followed by a survey of the library contents
from several different vantage points. CAS Registry Numbers are used
to assess ToxCast library coverage of important toxicity, regulatory,
and exposure inventories. Structure-based representations of ToxCast
chemicals are then used to compute physicochemical properties, substructural
features, and structural alerts for toxicity and biotransformation.
Cheminformatics approaches using these varied representations are
applied to defining the boundaries of HTS testability, evaluating
chemical diversity, and comparing the ToxCast library to potential
target application inventories, such as used in EPA’s Endocrine
Disruption Screening Program (EDSP). Through several examples, the
ToxCast chemical library is demonstrated to provide comprehensive
coverage of the knowledge domains and target inventories of potential
interest to EPA. Furthermore, the varied representations and approaches
presented here define local chemistry domains potentially worthy of
further investigation (e.g., not currently covered in the testing
library or defined by toxicity “alerts”) to strategically
support data mining and predictive toxicology modeling moving forward