Use of High Throughput Assays and Computational Tools; Endocrine Disruptor Screening Program; Notice of Availability and Opportunity for Comment, 35350-35355 [2015-15182]
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Authority: 44 U.S.C. 3501 et seq.
Dated: June 10, 2015.
James Jones,
Assistant Administrator, Office of Chemical
Safety and Pollution Prevention.
[FR Doc. 2015–14946 Filed 6–18–15; 8:45 am]
BILLING CODE 6560–50–P
ENVIRONMENTAL PROTECTION
AGENCY
[EPA–HQ–OPPT–2015–0305; FRL–9928–69]
Use of High Throughput Assays and
Computational Tools; Endocrine
Disruptor Screening Program; Notice
of Availability and Opportunity for
Comment
Environmental Protection
Agency (EPA).
ACTION: Notice.
AGENCY:
This document describes how
EPA is planning to incorporate an
alternative scientific approach to screen
chemicals for their ability to interact
with the endocrine system. This will
improve the Agency’s ability to fulfill its
statutory mandate to screen pesticide
chemicals and other substances for their
ability to cause adverse effects by their
interaction with the endocrine system.
The approach incorporates validated
high throughput assays and a
computational model and, based on
current research, can serve as an
alternative for some of the current
assays in the Endocrine Disruptor
Screening Program (EDSP) Tier 1
battery. EPA has partial screening
results for over 1800 chemicals that
have been evaluated using high
throughput assays and a computational
model for the estrogen receptor
pathway. In the future, EPA anticipates
that additional alternative methods will
be available for EDSP chemical
screening based on further
advancements of high throughput assays
and computational models for other
endocrine pathways. Use of these
alternative methods will accelerate the
pace of screening, decrease costs, and
reduce animal testing. In addition, this
approach advances the goal of providing
sensitive, specific, quantitative, and
SUMMARY:
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efficient screening using alternative test
methods to some assays in the Tier 1
battery to protect human health and the
environment.
DATES: Comments must be received on
or before August 18, 2015.
ADDRESSES: Submit your comments,
identified by docket identification (ID)
number EPA–HQ–OPPT–2015–0305, by
one of the following methods:
• Federal eRulemaking Portal: https://
www.regulations.gov. Follow the online
instructions for submitting comments.
Do not submit electronically any
information you consider to be
Confidential Business Information (CBI)
or other information whose disclosure is
restricted by statute.
• Mail: Document Control Office
(7407M), Office of Pollution Prevention
and Toxics (OPPT), Environmental
Protection Agency, 1200 Pennsylvania
Ave. NW., Washington, DC 20460–0001.
• Hand Delivery: To make special
arrangements for hand delivery or
delivery of boxed information, please
follow the instructions at https://
www.epa.gov/dockets/contacts.html.
Additional instructions on
commenting or visiting the docket,
along with more information about
dockets generally, is available at https://
www.epa.gov/dockets.
FOR FURTHER INFORMATION CONTACT: For
technical information contact: Jane
Robbins, Office of Science Coordination
and Policy (OSCP), Office of Chemical
Safety and Pollution Prevention,
Environmental Protection Agency, 1200
Pennsylvania Ave. NW., Washington,
DC 20460–0001; telephone number:
(202) 564–6625; email address:
robbins.jane@epa.gov.
For general information contact: The
TSCA-Hotline, ABVI-Goodwill, 422
South Clinton Ave., Rochester, NY
14620; telephone number: (202) 554–
1404; email address: TSCA-Hotline@
epa.gov.
SUPPLEMENTARY INFORMATION:
I. General Information
A. Does this action apply to me?
This action is directed to the public
in general, and may be of interest to a
wide range of stakeholders including
those interested in endocrine testing of
chemicals (including pesticides), and
the EDSP in general. Since others also
may be interested, the Agency has not
attempted to describe all the specific
entities that may be affected by this
action.
B. What is the agency authority for
taking this action?
The EDSP is established under section
408(p) of the Federal Food, Drug and
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Cosmetic Act (FFDCA), 21 U.S.C.
346a(p). Section 408(p)(1) requires EPA
‘‘to develop a screening program, using
appropriate validated test systems and
other scientifically relevant information
to determine whether certain substances
may have an effect in humans that is
similar to an effect produced by a
naturally occurring estrogen, or such
other effects as [EPA] may designate.’’
[21 U.S.C. 346a(p)(1)]. Section 408(p)(2)
requires that the screening program be
implemented ‘‘after obtaining public
comment and review . . . by the
scientific advisory panel established
under section 25(d) of the Federal
Insecticide, Fungicide, and Rodenticide
Act. . .’’ [21 U.S.C. 346a(p)(2)].
This document describes the new
scientific methods that are available as
alternatives to some of the current EDSP
Tier 1 screening assays and solicits
public comment on EPA’s plan to use
these alternative approaches to screen
chemicals for their ability to interact
with the endocrine system. The
approach described in this document is
not binding on either EPA or any
outside parties, and EPA may depart
from the approach presented in this
document where circumstances warrant
and without prior notice.
C. What action is the agency taking?
This document describes and solicits
comments on how EPA is planning to
incorporate scientific advancements and
tools into the EDSP. The adoption of
scientific advancements into the EDSP
has been underway and part of the
public dialogue about EDSP for several
years. As EPA has consistently
indicated, the Agency intends to
continue to incorporate in the EDSP
new methods involving high throughput
assays and computational toxicology.
Also, EPA has identified a universe of
approximately 10,000 chemicals as
potential candidates for screening and
testing under the EDSP (Ref. 1). This
approach is expected to accelerate the
pace of screening, add efficiencies,
decrease costs, and reduce animal
testing.
EPA is planning to incorporate the
partial screening results from validated
high throughput assays and
computational models as an alternative
to data from some of the current assays
in the EDSP Tier 1 screening battery.
Currently, EPA has partial screening
results for over 1800 chemicals that
have been evaluated using the high
throughput assays and computational
model for the estrogen receptor
pathway.
The use of high-throughput assays
and computational models for EDSP
screening is an initial step in EPA’s
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integration of 21st-century integrated
assessment and testing approaches
broadly, beyond EDSP, across a wide
range of chemicals related to regulatory
and non-regulatory decisions made in
programs under the Agency’s purview
(Ref. 2). Much of the knowledge gained
in using these approaches for EDSP
screening will be useful in applying
high throughput assays and
computational models to thousands of
chemicals across many toxicological
endpoints and exposure scenarios.
D. What should I consider as I prepare
my Comments for EPA?
1. Submitting CBI. Do not submit this
information to EPA through
regulations.gov or email. Clearly mark
the part or all of the information that
you claim to be CBI. For CBI
information in a disk or CD–ROM that
you mail to EPA, mark the outside of the
disk or CD–ROM as CBI and then
identify electronically within the disk or
CD–ROM the specific information that
is claimed as CBI. In addition to one
complete version of the comment that
includes information claimed as CBI, a
copy of the comment that does not
contain the information claimed as CBI
must be submitted for inclusion in the
public docket. Information so marked
will not be disclosed except in
accordance with procedures set forth in
40 CFR part 2.
2. Tips for preparing your comments.
When preparing and submitting your
comments, see the commenting tips at
https://www.epa.gov/dockets/
comments.html.
II. Background
A. What is the Endocrine Disruptor
Screening Program (EDSP)?
The Food Quality Protection Act
(FQPA) of 1996 amended FFDCA to
require EPA ‘‘to develop a screening
program, using appropriate validated
test systems and other scientifically
relevant information, to determine
whether certain substances may have an
effect in humans that is similar to an
effect produced by a naturally occurring
estrogen, or such other effects as [EPA]
may designate’’ (21 U.S.C. 346a(p)(1)).
Also in 1996, the Agency chartered the
Endocrine Disruptor Screening and
Testing Advisory Committee (EDSTAC),
under the provisions of the Federal
Advisory Committee Act (FACA) (5
U.S.C. App. 2, section 9(c)), to provide
advice on developing an endocrine
disruptor screening program (Ref. 3).
The EDSTAC was comprised of
members representing the commercial
chemical and pesticides industries,
Federal and State agencies, worker
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protection and labor organizations,
environmental and public health
groups, and research scientists. EDSTAC
recommended that EPA’s program
address both potential human and
wildlife effects; examine effects on
estrogen, androgen, and thyroid
hormone-related processes; and include
non-pesticide chemicals, contaminants,
and mixtures in addition to pesticide
chemicals (Ref. 2).
In 1998, based on the EDSTAC
recommendations, EPA established the
EDSP using a two-tiered approach (Ref.
4). The purpose of Tier 1 (referred to as
‘‘screening’’) is to identify substances
that have potential biological activity
(‘‘bioactivity’’) in the estrogen,
androgen, or thyroid hormone pathways
using a battery of assays. The purpose
of Tier 2 (referred to as ‘‘testing’’) is to
identify and establish a dose-response
relationship for any adverse effects that
might result from the endocrine
bioactivity identified through the Tier 1
assays. The ultimate purpose of the
EDSP is to provide information to the
Agency that will allow the Agency to
evaluate any possible endocrine effects
associated with the use of a chemical
and take appropriate steps to mitigate
any related risks to ensure protection of
public health.
In 2009, the Agency issued test orders
requiring Tier 1 screening for 67
chemicals (‘‘List 1’’) (Ref. 5). Between
the time needed to review the
substantial volume of ‘‘other
scientifically relevant information’’
submitted by test order recipients to
satisfy selected screening assays, the
time and resources of industry spent
generating data, the time spent by the
Agency reviewing the information, and
the delays resulting from the limited
laboratory capacity for conducting many
of the Tier 1 assays and corresponding
time extension requests, the review of
the initial List 1 chemicals has taken
over four years and has imposed
significant burdens on test order
recipients and the agency. The Agency
is still finalizing the data evaluation
records and determinations concerning
which of the List 1 chemicals need
further Tier 2 testing. More information
on the EDSP history and the status of
current activities is available at https://
www.epa.gov/endo.
B. What is meant by ‘‘high throughput
assays and computational model’’?
High throughput assays are automated
methods that allow for a large number
of chemicals to be rapidly evaluated for
a specific type of bioactivity at the
molecular or cellular level. This
approach, which can help identify
compounds that may modulate specific
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biological pathways, was initially
developed by pharmaceutical
companies for drug discovery. The
results of these methods provide an
initial understanding of a biochemical
interaction or possible role of a
chemical in a given biological process.
In vitro high throughput assays are
usually conducted using a microtiter
plate: a plate containing a grid with a
large number of small divots called
‘‘wells.’’ The wells contain chemical
and/or biological substrate (e.g., living
cells or proteins). Depending on the
nature of the experiment, changes can
be detected (e.g., color, fluorescence,
etc.) when the chemical is added to
indicate whether there is bioactivity.
High throughput microtiter plates
typically come in multiples of 96 wells
(96, 384, or 1536), so that through the
use of robotics, data processing and
control software, liquid handling
devices, and sensitive detection
methods, an extremely large number of
chemicals can be evaluated very
efficiently.
High throughput assays can be run for
a range of test chemical concentrations
and produce concentration-response
information representing the
relationship between chemical
concentration and bioactivity. The
concentration-response data from
multiple assays can be mathematically
integrated in a computational model of
a biological pathway, providing values
representative of a chemical’s
bioactivity in that pathway (e.g.,
estrogen receptor pathway). To reduce
non-specific results, the computational
model can use results from multiple
assays and technologies to predict
whether a chemical is truly bioactive in
the pathway being evaluated. The most
prominent cause of non-specific results
(activity in an assay that is likely not
due to bioactivity of the chemical in the
pathways) is cytotoxicity in cell-based
assays. In other cases, chemicals
influence the assays through a manner
dependent on the physics and chemistry
of the technology platform (i.e., ‘‘assay
interference’’).
C. What is ToxCastTM?
To improve efficiencies in screening
and testing chemicals, EPA scientists
are harnessing advances in molecular
and systems biology, chemistry,
toxicology, mathematics, and computer
technology. In doing this, they are
helping to revolutionize chemical
screening and safety testing based on
advances in computational toxicology.
A major part of this effort is the
Agency’s Toxicity Forecaster, or
ToxCastTM, which uses automated,
robotics-assisted high throughput assays
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to expose living cells or proteins to
chemicals and measure the results. The
high throughput assays produce
concentration-response information
representing the relationship between
chemical concentration and bioactivity.
These innovative methods have the
potential to quickly and efficiently
screen large numbers of chemicals and
other substances. ToxCastTM is part of
EPA’s contribution to a federal research
collaboration called ‘‘Toxicity Testing in
the 21st Century’’, or ‘‘Tox21,’’ pooling
resources and expertise from EPA, the
National Institutes of Health and the
U.S. Food and Drug Administration to
use robotics for screening thousands of
chemicals for potential bioactivity (Ref.
6).
As part of EPA’s commitment to
gather and share its chemical data
openly and clearly, all ToxCastTM
chemical data are publicly available
through user-friendly web applications
called the interactive Chemical Safety
for Sustainability (iCSS) and EDSP21
dashboards (Refs. 7 and 8). The EDSP21
and iCSS dashboards provide accessible
portals for users to search and query the
ToxCastTM chemical data. Users can
review chemicals and data of interest, as
well as export the information. Making
ToxCastTM data available through the
dashboards creates an environment that
encourages external stakeholder
interactions identifying potential issues,
concerns, and suggesting improvements.
D. What is meant by the ToxCastTM ER
Model for bioactivity?
The ToxCastTM ER Model for
bioactivity (‘‘ER Model’’) includes data
from 18 estrogen receptor (ER) high
throughput assays from ToxCastTM that
detect multiple events in the receptor
pathway. The ER Model also includes a
computational module that integrates
the assay data to produce a value for ER
agonist and antagonist bioactivity for
each chemical (Ref. 9). An ER agonist
binds and activates the receptor, and an
antagonist binds and blocks activation.
These 18 high throughput assays
measure bioactivity at different sites
along the ER pathway including
receptor binding, receptor dimerization,
chromatin binding of the mature
transcription factor, gene transcription
and changes in estrogen-receptor growth
kinetics. Bioactivity (i.e., response) is
measured using various detection
methods (e.g., fluorescence, etc.) across
a range of concentrations to examine
potential concentration-response
relationships, including no change
across concentrations indicating no
bioactivity. Concentration-response
relationships for each assay are
mathematically integrated in the ‘‘ER
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Model’’ to quantify bioactivity from
multiple assays. The computational
model integrates the results of each of
the 18 ER assays as an area under the
curve (AUC) for ER agonist or antagonist
bioactivity for each chemical. The
bioactivity values generally range from
0 to 1 for each chemical, with 0
indicating no bioactivity and 1
approximating the positive reference
chemical (e.g., estradiol for ER
agonism).
In order to validate the ER Model,
ToxCastTM data have been collected and
reviewed on over 1800 chemicals,
including ER reference agonists and
antagonists (Ref. 10). ER agonist and
antagonist bioactivity scores from the
‘‘ER Model’’ compare very well with
reported bioactivity of reference
chemicals across a range of structures
and potencies. Of the over 1800
chemicals tested, over 1700 chemicals
had very low or no detectable ER
bioactivity (Ref. 10). The ‘‘ER Model’’
bioactivity scores were validated by
comparing the scores to 45 reference
chemicals, equivalent to a performancebased approach to validation. EPA also
compared ‘‘ER Model’’ results to a
database of curated uterotrophic studies
published in peer-reviewed literature.
ER agonist bioactivity scores accurately
predicted in vivo ER agonist activity for
a large set (∼150) of chemicals with
uterotrophic data (Refs. 9 and 11). The
validation of the ‘‘ER Model’’ as an
alternative screening method for three
current Tier 1 assays (ER binding, ER
transcriptional activation (ERTA), and
uterotrophic) was peer reviewed by the
Federal Insecticide, Fungicide, and
Rodenticide Act (FIFRA) Scientific
Advisory Panel (SAP) in December 2014
(Refs. 9 and 11). The FIFRA SAP fully
endorsed the use of these alternatives
for the ER binding and ERTA assays;
however, there was not consensus
among panel members on the use of the
‘‘ER Model’’ as an alternative for the
uterotrophic assay (Ref. 11). In response
to the concerns raised by the FIFRA
SAP, EPA has published a paper
clarifying the relationship between ‘‘ER
Model’’ bioactivity and uterotrophic
results, and illustrating that a
uterotrophic assay would provide no
added value if ‘‘ER Model’’ data are
available (Ref. 12). Based on these
findings, EPA concludes that ‘‘ER
Model’’ data are sufficient to satisfy the
Tier 1 ER binding, ERTA and
uterotrophic assay requirements. The
Agency intends to build on the
performance-based validation approach
presented at the December 2014 FIFRA
SAP expanding this approach to include
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pathway.
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III. Using High Throughput Assays and
Computational Models for Screening
A. How Will ToxCastTM data be used for
screening in the EDSP?
The ability to screen chemicals
rapidly for bioactivity in several
endocrine pathways, and reducing the
use of animals in testing, have been
EDSP goals since 1998, when the
program was first adopted (Ref. 4). As
previously noted, when the first Tier 1
orders (for List 1 chemicals) were issued
in 2009, EPA had not confirmed the
reliability and relevance of the
ToxCastTM results so that they could be
cited as ‘‘other scientifically relevant
information’’ to satisfy the Tier 1 ER
binding, ERTA, and uterotrophic assays
(Ref. 13). However, since that time, EPA
has reached a critical juncture,
determining that the science has
progressed such that reevaluation of
EPA’s earlier position is warranted.
Based on scientific advances, EPA
intends to implement the use of high
throughput assays and computational
models to evaluate, and to a significant
extent, screen chemicals. The in vitro
high throughput and computational
model alternatives provide an accurate
quantitative measure of specific
endocrine pathway bioactivity and
mechanisms. The current Tier 1 battery
includes animal-based assays that do
not clearly identify or differentiate
pathways and mechanisms. Specifically,
the current Tier 1 ER binding, ERTA
and uterotrophic assays do not provide
both estrogen agonist and antagonist
activity and animals are required to
conduct the ER binding and
uterotrophic assays.
EPA is planning to adopt in vitro high
throughput assays and computational
models for detecting and measuring ER
agonist and antagonist bioactivity as an
alternative for three current Tier 1
assays: 1) ER binding in vitro assay (Ref.
14); 2) ER transcriptional activation in
vitro assay (ERTA) (Ref. 15); and 3) in
vivo uterotrophic assay (Refs. 16 and
17). EPA is also planning to accept
existing results for chemicals that have
been evaluated using the ToxCastTM
‘‘ER Model’’ for bioactivity. The
accompanying database contains the ER
agonist bioactivity and ER antagonist
bioactivity for over 1800 chemicals and
identifies those chemicals that are
pesticide active ingredients, pesticide
inert ingredients, and on EDSP Lists 1
or 2 (Ref. 10). This is a ‘‘living’’ database
that will continue to incorporate
bioactivity results for chemicals as they
become available. This database is
available at https://www.epa.gov/endo
and in the docket identified for this
document in a format that can be easily
reviewed and manipulated
electronically (Ref. 10). It is important,
however, not to equate a determination
of a chemical’s bioactivity from the ‘‘ER
Model’’ with a determination that a
chemical causes endocrine disruption.
The World Health Organization (WHO)/
International Programme on Chemical
Safety (IPCS) defines endocrine
disruption as being caused by ‘‘an
exogenous substance or mixture that
alters function(s) of the endocrine
system . . . and . . .consequently
causes adverse health effects in an intact
organism or its progeny, or
(sub)populations’’ (Ref. 18). Bioactivity
is an indicator that a chemical has the
potential to alter endocrine function,
but (1) whether the chemical actually
alters endocrine function and (2)
whether that altered function produces
an adverse outcome in an intact animal
cannot be determined without further
testing (i.e., Tier 2 testing).
The EDSP has been developed over
the past 19 years, and has demonstrated
that the current screening process may
take upwards of 5 years before a Tier 1
decision is available or Tier 2 test orders
are issued. In light of recent advances in
high throughput assays and
computational models, and advances
likely to come in the next two years, it
is prudent for the Agency to consider
new, rapid screening methods. The
availability of additional alternative
high throughput assays and
computational models in the near term
will allow EPA to screen more
chemicals in less time, involve fewer
animals, and cost less for everyone.
Furthermore, reconsideration of the
EDSP List 2 chemicals may be
appropriate since ‘‘ER Model’’ data are
available for many List 2 and other
chemicals (Refs. 10 and 19). Ongoing
use of high throughput assays and
computational models will address
thousands of chemicals in the future.
These advancements in the EDSP
screening program will not affect the
Estrogen Receptor (ER) Binding ..............................................................
Estrogen Receptor Transactivation (ERTA) .............................................
Uterotrophic ..............................................................................................
Female Rat Pubertal ................................................................................
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overall framework—i.e., the Tier 1
screening battery and Tier 2 testing
approach focused on estrogen, androgen
and thyroid pathways in humans and
wildlife remains unaffected. Instead, as
discussed above, EPA is planning to
adopt sensitive, specific, quantitative,
and efficient screening methods that
will rapidly screen many chemicals and
substantially decrease costs and animal
use and may be used as an alternative
to some EDSP Tier 1 screening assays.
Accordingly, EPA intends a future
recipient of an EDSP test order to be
able to satisfy the screening requirement
for ER, ERTA, and uterotrophic in one
of three ways: (1) cite existing
ToxCastTM ‘‘ER Model’’ for bioactivity
data as ‘‘other scientifically relevant
information’’ (where available); (2)
generate new data relying on the 18 ER
high throughput assays and the
ToxCastTM ‘‘ER Model’’ for bioactivity;
or (3) generate their own data using the
current Tier 1 ER binding, ERTA, and
uterotrophic assays.
B. How Does EPA intend to use high
throughput assays and computational
models for the EDSP in the future?
EPA believes that ongoing adoption of
alternative methods and technologies
will continue to advance EDSP
screening of chemicals for bioactivity in
the estrogen, androgen, and thyroid
pathways. EPA is continuing research
on the ‘‘ER Model’’ to determine if
ToxCastTM assays can provide
comparable information as that of the
Female Rat Pubertal and the Fish Short
Term Reproduction assays. In addition,
research continues on the ToxCastTM
‘‘AR Model’’ for bioactivity which, if
fully validated, may be considered as an
alternative (alone or with the ‘‘ER
Model’’) for the following current Tier 1
assays: AR binding, Male Rat Pubertal,
Hershberger, and Fish Short Term
Reproduction. Research is also
underway to develop steroidogenesis
ToxCastTM (STR) and thyroid (THY)
bioactivity models. Over time, the
Agency’s goal is to develop a set of
‘‘non-animal’’ high throughput assays
and computational bioactivity models as
an alternative to all of the assays in the
current Tier 1 screening battery. The
following table is intended to illustrate
the evolution of screening in the EDSP:
Alternative high throughput assays and computational model for EDSP
Tier 1 battery
Current EDSP Tier 1 battery of assays
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ER Model (alternative).
ER Model (alternative).
ER Model (alternative).
ER, STR , and thyroid (THY) Models (Future).
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Alternative high throughput assays and computational model for EDSP
Tier 1 battery
Current EDSP Tier 1 battery of assays
Male Rat Pubertal .....................................................................................
Androgen Receptor (AR) Binding .............................................................
Hershberger ..............................................................................................
Aromatase ................................................................................................
Steroidogenesis (STR) .............................................................................
Fish Short Term Reproduction .................................................................
Amphibian Metamorphosis .......................................................................
The table indicates combinations of
various alternative assays and models
that might overlap for evaluating
potential endocrine bioactivity of
chemicals. The in vitro high throughput
and computational model alternatives
provide a focused evaluation of the
mechanistic aspects of endocrine
pathways, thereby providing specific
and quantitative measures of bioactivity.
Several assays in the Tier 1 battery rely
on intact animals and identify
bioactivity in the multiple biological
pathways present. For this reason, the
specificity of the in vitro high
throughput and computational model
alternatives may be more informative of
specific endocrine pathway bioactivity.
The annual EDSP Comprehensive
Management Plan and future FIFRA
SAP meetings are opportunities for
staying informed on EPA’s scientific
progress on the evolution of Tier 1
screening in the EDSP. For information,
visit EPA’s Web site (https://
www.epa.gov/endo) or sign-up to
receive announcements go to (https://
www.epa.gov/endo/pubs/
assayvalidation/listserv.htm).
asabaliauskas on DSK5VPTVN1PROD with NOTICES
IV. Issues for Comment
In connection with EPA’s stated
intention to use the scientific tools
discussed in this Notice as alternatives
to some of the current EDSP Tier 1
screening assays, EPA is specifically
seeking public comment on the
following:
1. The use of the ToxCastTM ‘‘ER
Model’’ for bioactivity as an alternative
method for the current ER binding and
ERTA Tier 1 screening assays.
2. The use of the ToxCastTM ‘‘ER
Model’’ for bioactivity as an alternative
method for the current uterotrophic Tier
1 screening assay.
3. The use of results from the
ToxCastTM ‘‘ER Model’’ for bioactivity
on over 1800 chemicals as partial
screening for the estrogen receptor
pathway.
V. References
The following is a listing of the
documents that are specifically
referenced in this document. The docket
includes these documents and other
VerDate Sep<11>2014
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AR, STR, and THY Models (Future).
AR Model (Future).
AR Model (Future).
STR Model (Future).
STR Model (Future).
ER, AR, and STR Models (Future).
THY Model (Future).
information considered by EPA,
including documents that are referenced
within the documents that are included
in the docket, even if the referenced
document is not physically located in
the docket. For assistance in locating
these other documents, please consult
the technical person listed under FOR
FURTHER INFORMATION CONTACT.
1. U.S. EPA. Endocrine Disruptor Screening
Program; Universe of Chemicals and
General Validation Principles. November
2012. Available at https://www.epa.gov/
endo/pubs/edsp_chemical_universe_
and_general_validations_white_paper_
11_12.pdf.
2. U.S. EPA. Endocrine Disruptor Screening
Program for the 21st Century: (EDSP21
Work Plan); The Incorporation of In
Silico Models and In Vitro High
Throughput Assays in the Endocrine
Disruptor Screening Program (EDSP) for
Prioritization and Screening; Summary
Overview. A Part of the EDSP
Comprehensive Management Plan.
September 30, 2011. Available at https://
www.epa.gov/endo/pubs/edsp21_work_
plan_summary%20_overview_final.pdf.
3. U.S. EPA. Endocrine Disruptor Screening
and Testing Advisory Committee
(EDSTAC); Final Report. August 1998.
Available at https://www.epa.gov/endo/
pubs/edspoverview/finalrpt.htm.
4. U.S. EPA. Endocrine Disruptor Screening
Program; Proposed Statement of Policy;
Notice. Federal Register (63 FR 71542,
December 28, 1998) (FRL–6052–9).
5. U.S. EPA. Endocrine Disruptor Screening
Program; Tier 1 Screening Order
Issuance Announcement; Notice. Federal
Register (74 FR 54422, October 21, 2009)
(FRL–8434–8).
6. U.S. EPA. Office of Research and
Development (ORD); Description of
Computational Toxicology Research
Program. Available at https://epa.gov/
ncct.
7. U.S. EPA. Interactive Chemical Safety for
Sustainability (iCSS) Dashboard, Version
0.5. Available at https://actor.epa.gov/
dashboard.
8. U.S. EPA. EDSP21 Dashboard. Available at
https://actor.epa.gov/edsp21.
9. U.S. EPA. Integrated Bioactivity and
Exposure Ranking: A Computational
Approach for the Prioritization and
Screening of Chemicals in the Endocrine
Disruptor Screening Program. December
2014. Docket ID No. EPA–HQ–OPP–
2014–0614–0003. Available at https://
www.regulations.gov/
#!documentDetail;D=EPA–HQ–OPP–
PO 00000
Frm 00056
Fmt 4703
Sfmt 4703
2014–0614–0003.
10. U.S. EPA. Endocrine Disruptor Screening
Program (EDSP); Estrogen Receptor
Bioactivity Based on ToxCa TM ‘‘ER
Model.’’ June 1, 2015. Available at https://
www.epa.gov/endo.
11. U.S. EPA. FIFRA SAP Minutes No. 2015–
01. FIFRA SAP Meeting on the Integrated
Bioactivity and Exposure-Based
Prioritization and Screening, held
December 2–4, 2014. Docket ID No.
EPA–HQ–OPP–2014–0614–0029. March
2, 2015. Available at https://
www.epa.gov/scipoly/sap/meetings/
2014/december/120214minutes.pdf.
12. Browne, P., Judson, R.S., Casey, W.,
Kleinstreuer, N., Thomas, R.S. Screening
Chemicals For Estrogen Receptor
Bioactivity Using A Computational
Model. Manuscript accepted for
publication. Environ. Sci. Technol. June
12, 2015. Available in the docket and
electronically at https://pubs.acs.org/
journal/esthag.
13. U.S. EPA. Endocrine Disruptor Screening
Program; Policies and Procedures for
Initial Screening; Notice. Federal
Register (74 FR 17560, April 15, 2009)
(FRL–8399–9). Note: the status and
progress of all List 1 Tier 1 orders are
available at https://www.epa.gov/endo/
pubs/toresources/index.htm.
14. U.S. EPA. Endocrine Disruptor Screening
Program Test Guidelines; OPPTS
890.1250: Estrogen Receptor Binding
Assay Using Rat Uterine Cytosol (ER–
RUC). October 2009. EPA 740–C–09–005.
Available at https://www.epa.gov/ocspp/
pubs/frs/publications/Test_Guidelines/
series890.htm.
15.U.S.EPA. Endocrine Disruptor Screening
Program Test Guidelines; OPPTS
890.1300: Estrogen Receptor
Transcriptional Activation (Human Cell
Line (HeLa-9903)). October 2009. EPA
740–C–09–006. Available at https://
www.epa.gov/ocspp/pubs/frs/
publications/Test_Guidelines/
series890.htm.
16.U.S.EPA. Endocrine Disruptor Screening
Program Test Guidelines; OPPTS
890.1600: Uterotrophic Assay. October
2009. EPA 740–C–09–0010. Available at
https://www.epa.gov/ocspp/pubs/frs/
publications/Test_Guidelines/
series890.htm.
17. Organization of Economic Co-operation
and Development (OECD). Test
Guideline No. 440:Uterotrophic Bioassay
in Rodents: A short-term screening test
for oestrogenic properties. OECD
Guidelines for the Testing of Chemicals,
Section 4, OECD Publishing, Paris. DOI:
https://dx.doi.org/10.1787/
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Federal Register / Vol. 80, No. 118 / Friday, June 19, 2015 / Notices
9789264067417-en.
18. World Health Organization (WHO)/
International Programme on Chemical
Safety (IPCS). Global Assessment of the
State-of-the-Science of Endocrine
Disruptors. WHO/IPCS/EDC/02.2. 2002.
Available at https://www.who.int/ipcs/
publications/new_issues/endocrine_
disruptors/en.
19. U.S. EPA. Endocrine Disruptor Screening
Program; Final Second List of Chemicals
and Substances for Tier 1 Screening;
Notice. Federal Register (78 FR 35922,
June 14, 2013) (FRL–9375–8). Available
at https://www.gpo.gov/fdsys/pkg/FR–
2013–06–14/pdf/2013–14232.pdf.
Authority: 21 U.S.C. 346a(p).
Dated: June 11, 2015.
James J. Jones,
Assistant Administrator, Office of Chemical
Safety and Pollution Prevention.
[FR Doc. 2015–15182 Filed 6–18–15; 8:45 am]
BILLING CODE 6560–50–P
ENVIRONMENTAL PROTECTION
AGENCY
[EPA–HQ–SFUND–2006–0361; FRL—9929–
32–OSWER]
Proposed Information Collection
Request; Comment Request; Trade
Secret Claim Submissions under the
Emergency Planning and Community
Right-to-Know Act.
Environmental Protection
Agency (EPA).
ACTION: Notice.
AGENCY:
The Environmental Protection
Agency (EPA) is planning to submit an
information collection request (ICR),
‘‘Trade Secret Claims Submitted under
the Emergency Planning and
Community Right-to-Know Act.’’ (EPA
ICR No. 1428.10, OMB Control No.
2050–0078) to the Office of Management
and Budget (OMB) for review and
approval in accordance with the
Paperwork Reduction Act. Before doing
so, EPA is soliciting public comments
on specific aspects of the proposed
information collection as described
below. This is a proposed extension of
the ICR, which is currently approved
through December 31, 2015. An Agency
may not conduct or sponsor and a
person is not required to respond to a
collection of information unless it
displays a currently valid OMB control
number.
DATES: Comments must be submitted on
or before August 18, 2015.
ADDRESSES: Submit your comments,
referencing Docket ID No. EPA–HQ–
SFUND–2006–0361, online using
www.regulations.gov (our preferred
method), by email to
asabaliauskas on DSK5VPTVN1PROD with NOTICES
SUMMARY:
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19:33 Jun 18, 2015
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superfund.docket@epa.gov, or by mail
to: EPA Docket Center, Environmental
Protection Agency, Mail Code 28221T,
1200 Pennsylvania Ave. NW.,
Washington, DC 20460.
EPA’s policy is that all comments
received will be included in the public
docket without change including any
personal information provided, unless
the comment includes profanity, threats,
information claimed to be Confidential
Business Information (CBI) or other
information whose disclosure is
restricted by statute.
FOR FURTHER INFORMATION CONTACT: Sicy
Jacob, Office of Emergency
Management, Mail Code 5104A,
Environmental Protection Agency, 1200
Pennsylvania Ave. NW., Washington,
DC 20460; telephone number: (202)
564–8019; fax number: (202) 564–2620;
email address: jacob.sicy@epa.gov.
SUPPLEMENTARY INFORMATION:
Supporting documents which explain in
detail the information that the EPA will
be collecting are available in the public
docket for this ICR. The docket can be
viewed online at www.regulations.gov
or in person at the EPA Docket Center,
WJC West, Room 3334, 1301
Constitution Ave. NW., Washington,
DC. The telephone number for the
Docket Center is 202–566–1744. For
additional information about EPA’s
public docket, visit https://www.epa.gov/
dockets.
Pursuant to section 3506(c)(2)(A) of
the PRA, EPA is soliciting comments
and information to enable it to: (i)
evaluate whether the proposed
collection of information is necessary
for the proper performance of the
functions of the Agency, including
whether the information will have
practical utility; (ii) evaluate the
accuracy of the Agency’s estimate of the
burden of the proposed collection of
information, including the validity of
the methodology and assumptions used;
(iii) enhance the quality, utility, and
clarity of the information to be
collected; and (iv) minimize the burden
of the collection of information on those
who are to respond, including through
the use of appropriate automated
electronic, mechanical, or other
technological collection techniques or
other forms of information technology,
e.g., permitting electronic submission of
responses. EPA will consider the
comments received and amend the ICR
as appropriate. The final ICR package
will then be submitted to OMB for
review and approval. At that time, EPA
will issue another Federal Register
notice to announce the submission of
the ICR to OMB and the opportunity to
submit additional comments to OMB.
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35355
Abstract: This information collection
request pertains to trade secrecy claims
submitted under Section 322 of the
Emergency Planning and Community
Right-to-Know Act of 1986 (EPCRA).
EPCRA contains provisions requiring
facilities to report to State and local
authorities, and EPA, the presence of
extremely hazardous substances
(Section 302), inventory of hazardous
chemicals (Sections 311 and 312) and
manufacture, process and use of toxic
chemicals (Section 313).
Section 322 of EPCRA allows a
facility to withhold the specific
chemical identity from these EPCRA
reports if the facility asserts a claim of
trade secrecy for that chemical identity.
The provisions in Section 322 establish
the requirements and procedures that
facilities must follow to request trade
secrecy treatment of chemical identities,
as well as the procedures for submitting
public petitions to the Agency for
review of the ‘‘sufficiency’’ of trade
secrecy claims.
Trade secrecy protection is provided
for specific chemical identities
contained in reports submitted under
each of the following: (1) Section 303
(d)(2)- Facility notification of changes
that have or are about to occur, (2)
Section 303 (d)(3)—Local Emergency
Planning Committee (LEPC) requests for
facility information to develop or
implement emergency plans, (3) Section
311—Material Safety Data Sheets
(MSDSs) submitted by facilities, or lists
of those chemicals submitted in place of
the MSDSs, (4) Section 312—Emergency
and hazardous chemical inventory
forms (Tier I and Tier II), and (5) Section
313 Toxic chemical release inventory
form.
Form Number: EPA Form 9510–1.
Respondents/affected entities: Entities
potentially affected by this action are
manufacturers or non-manufacturers
subject to reporting under Sections 303,
311/312 or 313 of the Emergency
Planning and Community Right-toKnow Act (EPCRA).
Respondent’s obligation to respond:
Mandatory if the respondents would
like to claim the chemical identity for
any of the chemicals as trade secret in
any of the reports required to be
submitted under EPCRA.
Estimated number of respondents:
332 (total).
Frequency of response: Annual for
claims submitted under EPCRA Sections
312 and 313.
Total estimated burden: 3,154 hours
(per year). Burden is defined at 5 CFR
1320.03(b).
Total estimated cost: $206,155 (per
year). No capital and operation and
E:\FR\FM\19JNN1.SGM
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Agencies
[Federal Register Volume 80, Number 118 (Friday, June 19, 2015)]
[Notices]
[Pages 35350-35355]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 2015-15182]
-----------------------------------------------------------------------
ENVIRONMENTAL PROTECTION AGENCY
[EPA-HQ-OPPT-2015-0305; FRL-9928-69]
Use of High Throughput Assays and Computational Tools; Endocrine
Disruptor Screening Program; Notice of Availability and Opportunity for
Comment
AGENCY: Environmental Protection Agency (EPA).
ACTION: Notice.
-----------------------------------------------------------------------
SUMMARY: This document describes how EPA is planning to incorporate an
alternative scientific approach to screen chemicals for their ability
to interact with the endocrine system. This will improve the Agency's
ability to fulfill its statutory mandate to screen pesticide chemicals
and other substances for their ability to cause adverse effects by
their interaction with the endocrine system. The approach incorporates
validated high throughput assays and a computational model and, based
on current research, can serve as an alternative for some of the
current assays in the Endocrine Disruptor Screening Program (EDSP) Tier
1 battery. EPA has partial screening results for over 1800 chemicals
that have been evaluated using high throughput assays and a
computational model for the estrogen receptor pathway. In the future,
EPA anticipates that additional alternative methods will be available
for EDSP chemical screening based on further advancements of high
throughput assays and computational models for other endocrine
pathways. Use of these alternative methods will accelerate the pace of
screening, decrease costs, and reduce animal testing. In addition, this
approach advances the goal of providing sensitive, specific,
quantitative, and efficient screening using alternative test methods to
some assays in the Tier 1 battery to protect human health and the
environment.
DATES: Comments must be received on or before August 18, 2015.
ADDRESSES: Submit your comments, identified by docket identification
(ID) number EPA-HQ-OPPT-2015-0305, by one of the following methods:
Federal eRulemaking Portal: https://www.regulations.gov.
Follow the online instructions for submitting comments. Do not submit
electronically any information you consider to be Confidential Business
Information (CBI) or other information whose disclosure is restricted
by statute.
Mail: Document Control Office (7407M), Office of Pollution
Prevention and Toxics (OPPT), Environmental Protection Agency, 1200
Pennsylvania Ave. NW., Washington, DC 20460-0001.
Hand Delivery: To make special arrangements for hand
delivery or delivery of boxed information, please follow the
instructions at https://www.epa.gov/dockets/contacts.html.
Additional instructions on commenting or visiting the docket, along
with more information about dockets generally, is available at https://www.epa.gov/dockets.
FOR FURTHER INFORMATION CONTACT: For technical information contact:
Jane Robbins, Office of Science Coordination and Policy (OSCP), Office
of Chemical Safety and Pollution Prevention, Environmental Protection
Agency, 1200 Pennsylvania Ave. NW., Washington, DC 20460-0001;
telephone number: (202) 564-6625; email address: robbins.jane@epa.gov.
For general information contact: The TSCA-Hotline, ABVI-Goodwill,
422 South Clinton Ave., Rochester, NY 14620; telephone number: (202)
554-1404; email address: TSCA-Hotline@epa.gov.
SUPPLEMENTARY INFORMATION:
I. General Information
A. Does this action apply to me?
This action is directed to the public in general, and may be of
interest to a wide range of stakeholders including those interested in
endocrine testing of chemicals (including pesticides), and the EDSP in
general. Since others also may be interested, the Agency has not
attempted to describe all the specific entities that may be affected by
this action.
B. What is the agency authority for taking this action?
The EDSP is established under section 408(p) of the Federal Food,
Drug and
[[Page 35351]]
Cosmetic Act (FFDCA), 21 U.S.C. 346a(p). Section 408(p)(1) requires EPA
``to develop a screening program, using appropriate validated test
systems and other scientifically relevant information to determine
whether certain substances may have an effect in humans that is similar
to an effect produced by a naturally occurring estrogen, or such other
effects as [EPA] may designate.'' [21 U.S.C. 346a(p)(1)]. Section
408(p)(2) requires that the screening program be implemented ``after
obtaining public comment and review . . . by the scientific advisory
panel established under section 25(d) of the Federal Insecticide,
Fungicide, and Rodenticide Act. . .'' [21 U.S.C. 346a(p)(2)].
This document describes the new scientific methods that are
available as alternatives to some of the current EDSP Tier 1 screening
assays and solicits public comment on EPA's plan to use these
alternative approaches to screen chemicals for their ability to
interact with the endocrine system. The approach described in this
document is not binding on either EPA or any outside parties, and EPA
may depart from the approach presented in this document where
circumstances warrant and without prior notice.
C. What action is the agency taking?
This document describes and solicits comments on how EPA is
planning to incorporate scientific advancements and tools into the
EDSP. The adoption of scientific advancements into the EDSP has been
underway and part of the public dialogue about EDSP for several years.
As EPA has consistently indicated, the Agency intends to continue to
incorporate in the EDSP new methods involving high throughput assays
and computational toxicology. Also, EPA has identified a universe of
approximately 10,000 chemicals as potential candidates for screening
and testing under the EDSP (Ref. 1). This approach is expected to
accelerate the pace of screening, add efficiencies, decrease costs, and
reduce animal testing.
EPA is planning to incorporate the partial screening results from
validated high throughput assays and computational models as an
alternative to data from some of the current assays in the EDSP Tier 1
screening battery. Currently, EPA has partial screening results for
over 1800 chemicals that have been evaluated using the high throughput
assays and computational model for the estrogen receptor pathway.
The use of high-throughput assays and computational models for EDSP
screening is an initial step in EPA's integration of 21st-century
integrated assessment and testing approaches broadly, beyond EDSP,
across a wide range of chemicals related to regulatory and non-
regulatory decisions made in programs under the Agency's purview (Ref.
2). Much of the knowledge gained in using these approaches for EDSP
screening will be useful in applying high throughput assays and
computational models to thousands of chemicals across many
toxicological endpoints and exposure scenarios.
D. What should I consider as I prepare my Comments for EPA?
1. Submitting CBI. Do not submit this information to EPA through
regulations.gov or email. Clearly mark the part or all of the
information that you claim to be CBI. For CBI information in a disk or
CD-ROM that you mail to EPA, mark the outside of the disk or CD-ROM as
CBI and then identify electronically within the disk or CD-ROM the
specific information that is claimed as CBI. In addition to one
complete version of the comment that includes information claimed as
CBI, a copy of the comment that does not contain the information
claimed as CBI must be submitted for inclusion in the public docket.
Information so marked will not be disclosed except in accordance with
procedures set forth in 40 CFR part 2.
2. Tips for preparing your comments. When preparing and submitting
your comments, see the commenting tips at https://www.epa.gov/dockets/comments.html.
II. Background
A. What is the Endocrine Disruptor Screening Program (EDSP)?
The Food Quality Protection Act (FQPA) of 1996 amended FFDCA to
require EPA ``to develop a screening program, using appropriate
validated test systems and other scientifically relevant information,
to determine whether certain substances may have an effect in humans
that is similar to an effect produced by a naturally occurring
estrogen, or such other effects as [EPA] may designate'' (21 U.S.C.
346a(p)(1)). Also in 1996, the Agency chartered the Endocrine Disruptor
Screening and Testing Advisory Committee (EDSTAC), under the provisions
of the Federal Advisory Committee Act (FACA) (5 U.S.C. App. 2, section
9(c)), to provide advice on developing an endocrine disruptor screening
program (Ref. 3). The EDSTAC was comprised of members representing the
commercial chemical and pesticides industries, Federal and State
agencies, worker protection and labor organizations, environmental and
public health groups, and research scientists. EDSTAC recommended that
EPA's program address both potential human and wildlife effects;
examine effects on estrogen, androgen, and thyroid hormone-related
processes; and include non-pesticide chemicals, contaminants, and
mixtures in addition to pesticide chemicals (Ref. 2).
In 1998, based on the EDSTAC recommendations, EPA established the
EDSP using a two-tiered approach (Ref. 4). The purpose of Tier 1
(referred to as ``screening'') is to identify substances that have
potential biological activity (``bioactivity'') in the estrogen,
androgen, or thyroid hormone pathways using a battery of assays. The
purpose of Tier 2 (referred to as ``testing'') is to identify and
establish a dose-response relationship for any adverse effects that
might result from the endocrine bioactivity identified through the Tier
1 assays. The ultimate purpose of the EDSP is to provide information to
the Agency that will allow the Agency to evaluate any possible
endocrine effects associated with the use of a chemical and take
appropriate steps to mitigate any related risks to ensure protection of
public health.
In 2009, the Agency issued test orders requiring Tier 1 screening
for 67 chemicals (``List 1'') (Ref. 5). Between the time needed to
review the substantial volume of ``other scientifically relevant
information'' submitted by test order recipients to satisfy selected
screening assays, the time and resources of industry spent generating
data, the time spent by the Agency reviewing the information, and the
delays resulting from the limited laboratory capacity for conducting
many of the Tier 1 assays and corresponding time extension requests,
the review of the initial List 1 chemicals has taken over four years
and has imposed significant burdens on test order recipients and the
agency. The Agency is still finalizing the data evaluation records and
determinations concerning which of the List 1 chemicals need further
Tier 2 testing. More information on the EDSP history and the status of
current activities is available at https://www.epa.gov/endo.
B. What is meant by ``high throughput assays and computational model''?
High throughput assays are automated methods that allow for a large
number of chemicals to be rapidly evaluated for a specific type of
bioactivity at the molecular or cellular level. This approach, which
can help identify compounds that may modulate specific
[[Page 35352]]
biological pathways, was initially developed by pharmaceutical
companies for drug discovery. The results of these methods provide an
initial understanding of a biochemical interaction or possible role of
a chemical in a given biological process. In vitro high throughput
assays are usually conducted using a microtiter plate: a plate
containing a grid with a large number of small divots called ``wells.''
The wells contain chemical and/or biological substrate (e.g., living
cells or proteins). Depending on the nature of the experiment, changes
can be detected (e.g., color, fluorescence, etc.) when the chemical is
added to indicate whether there is bioactivity. High throughput
microtiter plates typically come in multiples of 96 wells (96, 384, or
1536), so that through the use of robotics, data processing and control
software, liquid handling devices, and sensitive detection methods, an
extremely large number of chemicals can be evaluated very efficiently.
High throughput assays can be run for a range of test chemical
concentrations and produce concentration-response information
representing the relationship between chemical concentration and
bioactivity. The concentration-response data from multiple assays can
be mathematically integrated in a computational model of a biological
pathway, providing values representative of a chemical's bioactivity in
that pathway (e.g., estrogen receptor pathway). To reduce non-specific
results, the computational model can use results from multiple assays
and technologies to predict whether a chemical is truly bioactive in
the pathway being evaluated. The most prominent cause of non-specific
results (activity in an assay that is likely not due to bioactivity of
the chemical in the pathways) is cytotoxicity in cell-based assays. In
other cases, chemicals influence the assays through a manner dependent
on the physics and chemistry of the technology platform (i.e., ``assay
interference'').
C. What is ToxCast\TM\?
To improve efficiencies in screening and testing chemicals, EPA
scientists are harnessing advances in molecular and systems biology,
chemistry, toxicology, mathematics, and computer technology. In doing
this, they are helping to revolutionize chemical screening and safety
testing based on advances in computational toxicology. A major part of
this effort is the Agency's Toxicity Forecaster, or ToxCast\TM\, which
uses automated, robotics-assisted high throughput assays to expose
living cells or proteins to chemicals and measure the results. The high
throughput assays produce concentration-response information
representing the relationship between chemical concentration and
bioactivity. These innovative methods have the potential to quickly and
efficiently screen large numbers of chemicals and other substances.
ToxCast\TM\ is part of EPA's contribution to a federal research
collaboration called ``Toxicity Testing in the 21st Century'', or
``Tox21,'' pooling resources and expertise from EPA, the National
Institutes of Health and the U.S. Food and Drug Administration to use
robotics for screening thousands of chemicals for potential bioactivity
(Ref. 6).
As part of EPA's commitment to gather and share its chemical data
openly and clearly, all ToxCast\TM\ chemical data are publicly
available through user-friendly web applications called the interactive
Chemical Safety for Sustainability (iCSS) and EDSP21 dashboards (Refs.
7 and 8). The EDSP21 and iCSS dashboards provide accessible portals for
users to search and query the ToxCast\TM\ chemical data. Users can
review chemicals and data of interest, as well as export the
information. Making ToxCast\TM\ data available through the dashboards
creates an environment that encourages external stakeholder
interactions identifying potential issues, concerns, and suggesting
improvements.
D. What is meant by the ToxCast\TM\ ER Model for bioactivity?
The ToxCast\TM\ ER Model for bioactivity (``ER Model'') includes
data from 18 estrogen receptor (ER) high throughput assays from
ToxCast\TM\ that detect multiple events in the receptor pathway. The ER
Model also includes a computational module that integrates the assay
data to produce a value for ER agonist and antagonist bioactivity for
each chemical (Ref. 9). An ER agonist binds and activates the receptor,
and an antagonist binds and blocks activation. These 18 high throughput
assays measure bioactivity at different sites along the ER pathway
including receptor binding, receptor dimerization, chromatin binding of
the mature transcription factor, gene transcription and changes in
estrogen-receptor growth kinetics. Bioactivity (i.e., response) is
measured using various detection methods (e.g., fluorescence, etc.)
across a range of concentrations to examine potential concentration-
response relationships, including no change across concentrations
indicating no bioactivity. Concentration-response relationships for
each assay are mathematically integrated in the ``ER Model'' to
quantify bioactivity from multiple assays. The computational model
integrates the results of each of the 18 ER assays as an area under the
curve (AUC) for ER agonist or antagonist bioactivity for each chemical.
The bioactivity values generally range from 0 to 1 for each chemical,
with 0 indicating no bioactivity and 1 approximating the positive
reference chemical (e.g., estradiol for ER agonism).
In order to validate the ER Model, ToxCast\TM\ data have been
collected and reviewed on over 1800 chemicals, including ER reference
agonists and antagonists (Ref. 10). ER agonist and antagonist
bioactivity scores from the ``ER Model'' compare very well with
reported bioactivity of reference chemicals across a range of
structures and potencies. Of the over 1800 chemicals tested, over 1700
chemicals had very low or no detectable ER bioactivity (Ref. 10). The
``ER Model'' bioactivity scores were validated by comparing the scores
to 45 reference chemicals, equivalent to a performance-based approach
to validation. EPA also compared ``ER Model'' results to a database of
curated uterotrophic studies published in peer-reviewed literature. ER
agonist bioactivity scores accurately predicted in vivo ER agonist
activity for a large set (~150) of chemicals with uterotrophic data
(Refs. 9 and 11). The validation of the ``ER Model'' as an alternative
screening method for three current Tier 1 assays (ER binding, ER
transcriptional activation (ERTA), and uterotrophic) was peer reviewed
by the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)
Scientific Advisory Panel (SAP) in December 2014 (Refs. 9 and 11). The
FIFRA SAP fully endorsed the use of these alternatives for the ER
binding and ERTA assays; however, there was not consensus among panel
members on the use of the ``ER Model'' as an alternative for the
uterotrophic assay (Ref. 11). In response to the concerns raised by the
FIFRA SAP, EPA has published a paper clarifying the relationship
between ``ER Model'' bioactivity and uterotrophic results, and
illustrating that a uterotrophic assay would provide no added value if
``ER Model'' data are available (Ref. 12). Based on these findings, EPA
concludes that ``ER Model'' data are sufficient to satisfy the Tier 1
ER binding, ERTA and uterotrophic assay requirements. The Agency
intends to build on the performance-based validation approach presented
at the December 2014 FIFRA SAP expanding this approach to include
[[Page 35353]]
other key events in the estrogen pathway.
III. Using High Throughput Assays and Computational Models for
Screening
A. How Will ToxCast\TM\ data be used for screening in the EDSP?
The ability to screen chemicals rapidly for bioactivity in several
endocrine pathways, and reducing the use of animals in testing, have
been EDSP goals since 1998, when the program was first adopted (Ref.
4). As previously noted, when the first Tier 1 orders (for List 1
chemicals) were issued in 2009, EPA had not confirmed the reliability
and relevance of the ToxCast\TM\ results so that they could be cited as
``other scientifically relevant information'' to satisfy the Tier 1 ER
binding, ERTA, and uterotrophic assays (Ref. 13). However, since that
time, EPA has reached a critical juncture, determining that the science
has progressed such that reevaluation of EPA's earlier position is
warranted. Based on scientific advances, EPA intends to implement the
use of high throughput assays and computational models to evaluate, and
to a significant extent, screen chemicals. The in vitro high throughput
and computational model alternatives provide an accurate quantitative
measure of specific endocrine pathway bioactivity and mechanisms. The
current Tier 1 battery includes animal-based assays that do not clearly
identify or differentiate pathways and mechanisms. Specifically, the
current Tier 1 ER binding, ERTA and uterotrophic assays do not provide
both estrogen agonist and antagonist activity and animals are required
to conduct the ER binding and uterotrophic assays.
EPA is planning to adopt in vitro high throughput assays and
computational models for detecting and measuring ER agonist and
antagonist bioactivity as an alternative for three current Tier 1
assays: 1) ER binding in vitro assay (Ref. 14); 2) ER transcriptional
activation in vitro assay (ERTA) (Ref. 15); and 3) in vivo uterotrophic
assay (Refs. 16 and 17). EPA is also planning to accept existing
results for chemicals that have been evaluated using the ToxCast\TM\
``ER Model'' for bioactivity. The accompanying database contains the ER
agonist bioactivity and ER antagonist bioactivity for over 1800
chemicals and identifies those chemicals that are pesticide active
ingredients, pesticide inert ingredients, and on EDSP Lists 1 or 2
(Ref. 10). This is a ``living'' database that will continue to
incorporate bioactivity results for chemicals as they become available.
This database is available at https://www.epa.gov/endo and in the docket
identified for this document in a format that can be easily reviewed
and manipulated electronically (Ref. 10). It is important, however, not
to equate a determination of a chemical's bioactivity from the ``ER
Model'' with a determination that a chemical causes endocrine
disruption. The World Health Organization (WHO)/International Programme
on Chemical Safety (IPCS) defines endocrine disruption as being caused
by ``an exogenous substance or mixture that alters function(s) of the
endocrine system . . . and . . .consequently causes adverse health
effects in an intact organism or its progeny, or (sub)populations''
(Ref. 18). Bioactivity is an indicator that a chemical has the
potential to alter endocrine function, but (1) whether the chemical
actually alters endocrine function and (2) whether that altered
function produces an adverse outcome in an intact animal cannot be
determined without further testing (i.e., Tier 2 testing).
The EDSP has been developed over the past 19 years, and has
demonstrated that the current screening process may take upwards of 5
years before a Tier 1 decision is available or Tier 2 test orders are
issued. In light of recent advances in high throughput assays and
computational models, and advances likely to come in the next two
years, it is prudent for the Agency to consider new, rapid screening
methods. The availability of additional alternative high throughput
assays and computational models in the near term will allow EPA to
screen more chemicals in less time, involve fewer animals, and cost
less for everyone. Furthermore, reconsideration of the EDSP List 2
chemicals may be appropriate since ``ER Model'' data are available for
many List 2 and other chemicals (Refs. 10 and 19). Ongoing use of high
throughput assays and computational models will address thousands of
chemicals in the future.
These advancements in the EDSP screening program will not affect
the overall framework--i.e., the Tier 1 screening battery and Tier 2
testing approach focused on estrogen, androgen and thyroid pathways in
humans and wildlife remains unaffected. Instead, as discussed above,
EPA is planning to adopt sensitive, specific, quantitative, and
efficient screening methods that will rapidly screen many chemicals and
substantially decrease costs and animal use and may be used as an
alternative to some EDSP Tier 1 screening assays. Accordingly, EPA
intends a future recipient of an EDSP test order to be able to satisfy
the screening requirement for ER, ERTA, and uterotrophic in one of
three ways: (1) cite existing ToxCast\TM\ ``ER Model'' for bioactivity
data as ``other scientifically relevant information'' (where
available); (2) generate new data relying on the 18 ER high throughput
assays and the ToxCast\TM\ ``ER Model'' for bioactivity; or (3)
generate their own data using the current Tier 1 ER binding, ERTA, and
uterotrophic assays.
B. How Does EPA intend to use high throughput assays and computational
models for the EDSP in the future?
EPA believes that ongoing adoption of alternative methods and
technologies will continue to advance EDSP screening of chemicals for
bioactivity in the estrogen, androgen, and thyroid pathways. EPA is
continuing research on the ``ER Model'' to determine if ToxCast\TM\
assays can provide comparable information as that of the Female Rat
Pubertal and the Fish Short Term Reproduction assays. In addition,
research continues on the ToxCast\TM\ ``AR Model'' for bioactivity
which, if fully validated, may be considered as an alternative (alone
or with the ``ER Model'') for the following current Tier 1 assays: AR
binding, Male Rat Pubertal, Hershberger, and Fish Short Term
Reproduction. Research is also underway to develop steroidogenesis
ToxCast\TM\ (STR) and thyroid (THY) bioactivity models. Over time, the
Agency's goal is to develop a set of ``non-animal'' high throughput
assays and computational bioactivity models as an alternative to all of
the assays in the current Tier 1 screening battery. The following table
is intended to illustrate the evolution of screening in the EDSP:
------------------------------------------------------------------------
Alternative high throughput
Current EDSP Tier 1 battery of assays assays and computational model
for EDSP Tier 1 battery
------------------------------------------------------------------------
Estrogen Receptor (ER) Binding......... ER Model (alternative).
Estrogen Receptor Transactivation ER Model (alternative).
(ERTA).
Uterotrophic........................... ER Model (alternative).
Female Rat Pubertal.................... ER, STR , and thyroid (THY)
Models (Future).
[[Page 35354]]
Male Rat Pubertal...................... AR, STR, and THY Models
(Future).
Androgen Receptor (AR) Binding......... AR Model (Future).
Hershberger............................ AR Model (Future).
Aromatase.............................. STR Model (Future).
Steroidogenesis (STR).................. STR Model (Future).
Fish Short Term Reproduction........... ER, AR, and STR Models
(Future).
Amphibian Metamorphosis................ THY Model (Future).
------------------------------------------------------------------------
The table indicates combinations of various alternative assays and
models that might overlap for evaluating potential endocrine
bioactivity of chemicals. The in vitro high throughput and
computational model alternatives provide a focused evaluation of the
mechanistic aspects of endocrine pathways, thereby providing specific
and quantitative measures of bioactivity. Several assays in the Tier 1
battery rely on intact animals and identify bioactivity in the multiple
biological pathways present. For this reason, the specificity of the in
vitro high throughput and computational model alternatives may be more
informative of specific endocrine pathway bioactivity.
The annual EDSP Comprehensive Management Plan and future FIFRA SAP
meetings are opportunities for staying informed on EPA's scientific
progress on the evolution of Tier 1 screening in the EDSP. For
information, visit EPA's Web site (https://www.epa.gov/endo) or sign-up
to receive announcements go to (https://www.epa.gov/endo/pubs/assayvalidation/listserv.htm).
IV. Issues for Comment
In connection with EPA's stated intention to use the scientific
tools discussed in this Notice as alternatives to some of the current
EDSP Tier 1 screening assays, EPA is specifically seeking public
comment on the following:
1. The use of the ToxCast\TM\ ``ER Model'' for bioactivity as an
alternative method for the current ER binding and ERTA Tier 1 screening
assays.
2. The use of the ToxCast\TM\ ``ER Model'' for bioactivity as an
alternative method for the current uterotrophic Tier 1 screening assay.
3. The use of results from the ToxCast\TM\ ``ER Model'' for
bioactivity on over 1800 chemicals as partial screening for the
estrogen receptor pathway.
V. References
The following is a listing of the documents that are specifically
referenced in this document. The docket includes these documents and
other information considered by EPA, including documents that are
referenced within the documents that are included in the docket, even
if the referenced document is not physically located in the docket. For
assistance in locating these other documents, please consult the
technical person listed under FOR FURTHER INFORMATION CONTACT.
1. U.S. EPA. Endocrine Disruptor Screening Program; Universe of
Chemicals and General Validation Principles. November 2012.
Available at https://www.epa.gov/endo/pubs/edsp_chemical_universe_and_general_validations_white_paper_11_12.pdf.
2. U.S. EPA. Endocrine Disruptor Screening Program for the 21st
Century: (EDSP21 Work Plan); The Incorporation of In Silico Models
and In Vitro High Throughput Assays in the Endocrine Disruptor
Screening Program (EDSP) for Prioritization and Screening; Summary
Overview. A Part of the EDSP Comprehensive Management Plan.
September 30, 2011. Available at https://www.epa.gov/endo/pubs/edsp21_work_plan_summary%20_overview_final.pdf.
3. U.S. EPA. Endocrine Disruptor Screening and Testing Advisory
Committee (EDSTAC); Final Report. August 1998. Available at https://www.epa.gov/endo/pubs/edspoverview/finalrpt.htm.
4. U.S. EPA. Endocrine Disruptor Screening Program; Proposed
Statement of Policy; Notice. Federal Register (63 FR 71542, December
28, 1998) (FRL-6052-9).
5. U.S. EPA. Endocrine Disruptor Screening Program; Tier 1 Screening
Order Issuance Announcement; Notice. Federal Register (74 FR 54422,
October 21, 2009) (FRL-8434-8).
6. U.S. EPA. Office of Research and Development (ORD); Description
of Computational Toxicology Research Program. Available at https://epa.gov/ncct.
7. U.S. EPA. Interactive Chemical Safety for Sustainability (iCSS)
Dashboard, Version 0.5. Available at https://actor.epa.gov/dashboard.
8. U.S. EPA. EDSP21 Dashboard. Available at https://actor.epa.gov/edsp21.
9. U.S. EPA. Integrated Bioactivity and Exposure Ranking: A
Computational Approach for the Prioritization and Screening of
Chemicals in the Endocrine Disruptor Screening Program. December
2014. Docket ID No. EPA-HQ-OPP-2014-0614-0003. Available at https://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPP-2014-0614-0003.
10. U.S. EPA. Endocrine Disruptor Screening Program (EDSP); Estrogen
Receptor Bioactivity Based on ToxCa \TM\ ``ER Model.'' June 1, 2015.
Available at https://www.epa.gov/endo.
11. U.S. EPA. FIFRA SAP Minutes No. 2015-01. FIFRA SAP Meeting on
the Integrated Bioactivity and Exposure-Based Prioritization and
Screening, held December 2-4, 2014. Docket ID No. EPA-HQ-OPP-2014-
0614-0029. March 2, 2015. Available at https://www.epa.gov/scipoly/sap/meetings/2014/december/120214minutes.pdf.
12. Browne, P., Judson, R.S., Casey, W., Kleinstreuer, N., Thomas,
R.S. Screening Chemicals For Estrogen Receptor Bioactivity Using A
Computational Model. Manuscript accepted for publication. Environ.
Sci. Technol. June 12, 2015. Available in the docket and
electronically at https://pubs.acs.org/journal/esthag.
13. U.S. EPA. Endocrine Disruptor Screening Program; Policies and
Procedures for Initial Screening; Notice. Federal Register (74 FR
17560, April 15, 2009) (FRL-8399-9). Note: the status and progress
of all List 1 Tier 1 orders are available at https://www.epa.gov/endo/pubs/toresources/index.htm.
14. U.S. EPA. Endocrine Disruptor Screening Program Test Guidelines;
OPPTS 890.1250: Estrogen Receptor Binding Assay Using Rat Uterine
Cytosol (ER-RUC). October 2009. EPA 740-C-09-005. Available at
https://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series890.htm.
15.U.S.EPA. Endocrine Disruptor Screening Program Test Guidelines;
OPPTS 890.1300: Estrogen Receptor Transcriptional Activation (Human
Cell Line (HeLa-9903)). October 2009. EPA 740-C-09-006. Available at
https://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series890.htm.
16.U.S.EPA. Endocrine Disruptor Screening Program Test Guidelines;
OPPTS 890.1600: Uterotrophic Assay. October 2009. EPA 740-C-09-0010.
Available at https://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series890.htm.
17. Organization of Economic Co-operation and Development (OECD).
Test Guideline No. 440:Uterotrophic Bioassay in Rodents: A short-
term screening test for oestrogenic properties. OECD Guidelines for
the Testing of Chemicals, Section 4, OECD Publishing, Paris. DOI:
https://dx.doi.org/10.1787/
[[Page 35355]]
9789264067417-en.
18. World Health Organization (WHO)/International Programme on
Chemical Safety (IPCS). Global Assessment of the State-of-the-
Science of Endocrine Disruptors. WHO/IPCS/EDC/02.2. 2002. Available
at https://www.who.int/ipcs/publications/new_issues/endocrine_disruptors/en.
19. U.S. EPA. Endocrine Disruptor Screening Program; Final Second
List of Chemicals and Substances for Tier 1 Screening; Notice.
Federal Register (78 FR 35922, June 14, 2013) (FRL-9375-8).
Available at https://www.gpo.gov/fdsys/pkg/FR-2013-06-14/pdf/2013-14232.pdf.
Authority: 21 U.S.C. 346a(p).
Dated: June 11, 2015.
James J. Jones,
Assistant Administrator, Office of Chemical Safety and Pollution
Prevention.
[FR Doc. 2015-15182 Filed 6-18-15; 8:45 am]
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