HESI:
Health and Environmental Sciences Institute
Syril D Pettit, MEM
Executive Director
Seminar for NIES
February 26, 2015
HESI
Washington, DC USA
www.hesiglobal.org
ILSI Health and
Environmental Sciences
Institute
HESI Mission
Create science-based solutions for a
sustainable, healthier world.
Accurate
and Efficient
Chemical
Risk
Assessment
Safe and
Effective
Medicines
Environmental
Quality and
Sustainability
Food Safety
Protecting
sensitive
populations
Sustaining critical
environments
Supporting ecological
and human safety of
essential food
resources
Promoting Discovery
Predicting and
Protecting Against
Adverse Effects
from Chronic
Exposures
The HESI Model:
Bridging Research to Application
Academic
& basic
research
sector
SAFETY &
INNOVATION FOR
HUMAN &
ENVIRONMENTAL
HEALTH
Patient
Advocates,
Foundations
&
NGOs
Industry
R&D
Government
Research
&
Regulation
90
University
&
Research
Centers
From 14
Countries
47
Government
Agencies
66
Corporate
Sponsors
Across multiple
sectors
From 12
Countries
14
Scientific
Committees
>70
Distinct
Projects
Impact via
Quality
Science
HESI achieves its mission via…
Scientific
Research
Millions in in-kind
research annually
Publication
Communication
& Translation
Tools
Training
Novel programs
with
interdisciplinary
and cross-sector
focus
Platforms for
Interaction
Active public-private partnership
We know the model works…
HESI’s scientific programs and publications have..
Influenced their approach to safety or risk
assessment decision-making; 70%
Influenced their level of confidence in the use of
particular technologies, markers, endpoints, or
analysis approaches; 80%
Directly and positively impacted
safety of patients and the environment
History of Success
Creating frameworks to
integrate data and
decision-making
Prioritize risks,
Protect
ecosystems and
their inhabitants
Resources
“The recommendations from these
From HESI’s First
Program…
early HESI-EPA workshops provided a
foundation upon which to develop a
mode of action (MOA) framework.
The IPCS defined the criteria for
accepting a MOA as adequate for
evaluating a specific tumor type in
animals…subsequent work
…determined how MOA studies can be
used to establish the relevance of
rodent tumors to humans.”
Integrating Alternatives
to Animal Testing for
Ecotox
Globally
recognized HESI
roadmaps to
guide
integration of data
and decisions.
Mode of Action
Risk / Safety
In vivo
In vitro
Toxicity
Innovating
Chemical Risk
Assessment
Matrix
QSAR/
TTC
Biomonitoring
Probabilistic
Deterministic
Minimal
Info
Assessing Adverse vs
Adaptive Transitions in
Toxicity Pathways
Exposure
Problem
Formulation
Conclude
Enhancing
AgChem
Safety
A spotlight
on one of
many…
US EPA Scientific and Technological
 Basis for OECD Guideline for
Achievement Award
Testing of Chemicals (443):
(Honorable Mention)
Extended One-Generation
Reproductive Toxicity Study
UK National Center for the
 Canine study requirement
Replacement, Refinement, and
dropped in EPA Pesticide
Reduction of Animals in Research
guidelines;
“Highly Commended Prize”
 Increased use of ADME to
enhance dose selection
Impact cited in 2 National Academy
reports
Enhancing
AgChem
Safety
Informing discovery
& decision-making
with new technologies
Toxicogenomics • First large scale TGx experimental
program, first public array/tox dbase
for Risk
• Led to adoption of data standards,
Assessment
genomic biomarkers
• Resource for strengths & limitations
of TgX use for safety
Transgenic Models for
Cancer Risk Assessment
• $33M collaborative effort
• Critical data on predictivity of
available transgenic models
• Data underpins current
guidelines on alternatives to
2 year mouse bioassay
• Improved prediction of safety
Translating from animal to human,
and back to improve predictive
safety
HESI Approach to Biomarkers
• Consensus on Safety or Translational Need
• Experimental Data
• Analysis & Publication
Nonclinical
cTn serum
assays
• Integration of Data & Context of Use
Nonclinical
Inhibin
Assays
Urinary
Renal
Protein
Biomarkers
MicroRNAs as
translation tox markers
Chemical Safety
Evaluation
Predictive models;
Risk assessment methodologies;
Sustainability
Capacity Building & Education
Predictive Models
Zebrafish &
multigenerational
epigenetics
Bioaccumulation:
In vitro method,
hepatic
clearance in trout
Utility of 2nd Species
for assessing
developmental
toxicity
Pig-A assay for
genotoxicity
Risk Assessment Methodologies
RISK21
AOP and
Ecotox
Sustainability
Capacity Building & Education
Scientist from the
following
organization’s
collaborate with
HESI on Chemical
Risk Assessment
research….
and many more!
The HESI Model:
Bridging Research to Application
Academic
& basic
research
sector
SAFETY &
INNOVATION FOR
HUMAN &
ENVIRONMENTAL
HEALTH
Patient
Advocates,
Foundations
&
NGOs
Industry
R&D
Government
Research
&
Regulation
Questions?
SYRIL D PETTIT
HESI EXECUTIVE DIRECTOR
SPETTIT@HESIGLOBAL.ORG
WWW.HESIGLOBAL.ORG
AYAKO TAKEI
HESI SCIENCE ADVISOR IN JAPAN
ATAKEI@HESIGLOBAL.ORG
With this framework
LET’S DO A DEEPER DIVE INTO SOME SPECIFIC PROGRAMS…
Mode of Action
Risk / Safety
In vivo
In vitro
Toxicity
RISK21
Matrix
QSAR/
TTC
Biomonitoring
Probabilistic
Deterministic
Minimal
Info
Exposure
Problem
Formulation
Conclude
Risk Assessment in the 21st Century (RISK21)
• MISSION: Bring applicable, accurate, and resource
appropriate approaches to the evolving world of human
health risk assessment
•
Convened experts from academia, industry, government
and other stakeholders
•
•
RISK21 involved > 120 scientists from Europe and USA
•
Revised current thinking about how to approach the
science and art of risk assessment
Developed a risk assessment approach that embraces
advances in scientific knowledge and methods
How is RISK21 Different?
• Think about the problem that needs to be
addressed; then select sources of information which
will have the most value
• RISK21 Principles:
–
–
–
–
Problem-formulation based
Exposure-driven
Prior knowledge
“Enough precision to make the decision”
• Provides a framework that is…
– Flexible
– Transparent
– Visual
Risk / Safety
Mode of Action
In vivo
4
In vitro
3
QSAR/
TTC
Toxicity
Biomonitoring
Probabilistic
Deterministic
Minimal
Info
2
Exposure
1
Problem
Formulation
Conclude
High
Toxicity range
0.001
Mod
0.01
0.1
1.0
10
Exposure range
Low
Estimate of Human Toxicity (mg/kg)
Plotting Ranges on the RISK21 Matrix
0.0001
0.001
Low
0.01
0.1
Mod
1
10
Estimate of Human Exposure (mg/kg)
100
High
Web-Tool beta version available: http://risk21.sciome.com/
Use of RISK21 Matrix
MOE set at 1:1
Benefits of the RISK21 Matrix
• Visual
• Effective risk communications tool
• Multiple applications:
– Priority setting
– Evaluation of data needs
– Evaluation of new use or release scenario
• Flexible and transparent
• Allows incorporation of new data when appropriate
• Can inform study design & resource allocation
Publications
Critical Reviews in Toxicology, 2014; 44(S3): 1–5
Critical Reviews in Toxicology, 2014; 44(S3): 6–16
Critical Reviews in Toxicology, 2014; 44(S3): 17–43
OPEN ACCESS!
RISK21: Next steps
• Additional publications on:
–
–
–
–
Case studies (chemicals in water and pyrethroid)
Cumulative risk
Exposure
In vitro to in vivo extrapolation
• Training seminars and additional case studies
• New projects on exposure and risk
For More Information on
HESI’s RISK21 Project, contact…
Dr. Michelle Embry
HESI Sr. Scientific Program Manager
membry@hesiglobal.org
www.hesiglobal.org
www.risk21.org
BIOACCUMULATION
OF CHEMICALS
Bioaccumulation Committee: Mission and Objectives
– To develop the tools needed for assessing
the potential bioaccumulation of organic
chemicals,
– To address how the various metrics used
to assess bioaccumulation can be
integrated to develop an overall weight of
evidence approach for deriving
assessment conclusions, and
– To partner with other groups involved in
research and improvements in
bioaccumulation methods and
assessment.
38
Key Drivers for B Work
•
National and international regulatory programs are focused
on identifying and controlling chemicals that are Persistent,
Bioaccumulative, and Toxic (PBT)
•
Stockholm Convention led to increased need for PBT
assessment
•
Bioaccumulation data are scarce (<3% of all chemicals
have data)
•
Animal testing reduction goals and acceptance of these
data
39
Various Bioaccumulation committee activities
• Development of an in vitro metabolism assay
• Development of a new model for chemical
transfer efficiency across the fish GI tract
• Bioaccumulation in terrestrial systems
• Benchmarking
• Trophic Magnification Factor studies: best
practices
• Risk assessment of ionizable organic
compounds
40
In vitro Fish Hepatic Metabolism OECD Project 3.13
A multi-site
laboratory ring trial
ILSI Health and
Environmental Sciences
Institute
Joint Research Centre
United States Environmental
Protection Agency
Bioaccumulation: in vitro metabolism
Accumulation of a chemical in an
organism is the result of absorption,
distribution, metabolism, and excretion
(ADME).
Loss via
gills
(k2)
Loss via
egestion
(kE)
Uptake via
food
Loss via
metabolism
(kMET)
Uptake via
gills
(k1)
•
BCF models based on log KOW are used to
screen chemicals for potential
bioaccumulative properties: Do not
account for “M” (i.e., biotransformation)!
•
Some QSAR models predict metabolism
rates based on chemical structure (incl. in
BCF/BAF™ programme of US EPA Episuite)
•
Only waterborne uptake
•
BCF = k1 / (k2 + kE + kMET)
In vitro methods?
Overall aim of the OECD 3.13
• Development of an OECD test guideline for in vitro
determination of hepatic biotransformation in fish
• Supports in silico determination of bioaccumulation
For this purpose:
• The performance of two in vitro methods based on
rainbow trout S9 and cryopreserved hepatocytes will be
evaluated within and across participating laboratories:
• Reproducibility
• Activity
• Ring trial involving seven laboratories and testing of six
chemicals
In vitro method: Application for Bioaccumulation
Calculate in vitro intrinsic clearance rate for the
parent chemical
Analyze
Extract parent compound
•Liver weight (g/kg)
•Hepatocellularity (cells/g liver)
•S9 protein content (protein/g liver)
Calculate in vivo intrinsic clearance rate (L/h/kg)
• Liver blood flow rate (L/d/kg)
• Binding corrections as appropriate
Quench reaction at
regular intervals
Calculate hepatic clearance (L/h/kg)
• Apparent volume of distribution (L/kg)
Incubate w/ test chemical
Calculate whole-fish metabolism rate constant kMET (1/d)
Combine kMET with estimates of k1, k2 and kE to simulate CFish and
predict BCF
Isolate liver S9 fraction or
hepatocytes
Fay et al, 2014; Nichols et al., 2013; Johanning et al, 2012;
Cowan-Ellsberry et al., 2008; Han et al., 2007
Chemical Selection
4-nnonylphenol
Pyrene
Cyclohexyl
salicylate
Fenthion
•
•
•
•
•
•
Methoxychlor
Deltamethrin
Range of log Kow (4 – 6)
High quality in vivo fish BCF data
Available in vitro data
Well-behaved
Analytical capabilities of participating laboratories
Range of chemical classes
Participating Laboratories
USEPA
DuPont
Dow
Givaudan
Fraunhofer
Procter &
Gamble
KJ Scientific
/ SCJ
Isolation of
biological
material
Incubations
Incubations
Incubations
Incubations
Incubations
Incubations
Analytical:
Pyrene,
Fenthion
Analytical:
4NP
Analytical:
Deltamethrin
Analytical:
Cyclohexyl
salicylate
Analytical:
Methoxychlor
Biological Material Characterization
• Each lot of biological material (11 lots of S9, 8 lots of hepatocytes)
characterized using prototypical substrates for:
• CYP1A
• Glutathione transferase (GST)
• Uridine 5'-diphospho-glucuronosyl transferase (UGT)
• Carboxyl esterase
• Each chemical will be run with the same lot for all labs
Status & Timeline
Completed
January 2015
February 2015
Participating laboratories
identified
Modification of SOPs
Optimization of test conditions
for all chemicals
Biological material isolated
Incubations with pyrene in all 6
laboratories
Start of incubations with
additional test chemicals
Biological material characterized
Analytical analysis of pyrene data
Test chemicals selected
Analytical laboratories identified
Study design optimized
SOPs developed
Biological material shipped
All incubations completed by 2Q 2015
Analytical completed by 3Q 2015
Test chemicals ordered
Pyrene conditions optimized
(positive control / pilot)
Finalization of SOPs
Development of data
spreadsheet
Final report by early 2016
Questions?
CONTACT INFORMATION:
Michelle Embry (membry@hesiglobal.org)
Sustainable
alternatives
51
Today’s Challenging Landscape

Increasing pressures to find more sustainable, safer alternatives
Regulatory Drivers: Replace Chemicals of Concern

REACH Authorization

California Safer Consumer Products Regulation

TSCA reform
Corporate Sustainability Initiatives

Greenhouse gas reduction

Energy conservation

Raw material preservation

Reduced hazard options
Customer/Consumer Drivers

Banned lists of chemicals

Ecolabel certifications
52
HESI Sustainable Alternatives
Committee - Formed in 2011

Mission: To evaluate and identify key elements/criteria
and tools to help trigger and guide the selection of safer,
sustainable alternatives while minimizing the likelihood of
regrettable substitutions.

Objective: The main objective is to develop practical,
problem-driven guidance on the conduct of alternative
chemical assessment.
 Workshop: 7-8 February 2013 at
NIEHS in North Carolina.
Small Pilot Project Highlights Opportunity to Improve Tools1
EXAMPLE: RESULTS OF CHEMICAL SCREENING TOOLS COMPARISON
SCREENING TOOL SCORE
GreenWercs GreenWercs GreenScreen GreenScreen
CHEMICAL NAME
Natural Chemical A
Natural Chemical B
Industrial Chemical A
Natural Chemical C
Industrial Chemical B
Industrial Chemical C
Industrial Chemical D
1Sponsored
(Walmart
(GreenScreen
Scoring Model)
Model)
(full
assessment)
(list translator)
0-5000
0-2500
(preferable); 5000(preferable);
15000
2500-6000
(acceptable);
(acceptable);
15000-20000
6000-8500 (avoid)
(avoid)
BM1 worst;
BM4 best
LT-1 = BM1; LTP1 = Possible BM1; (concern - # endpoints of
LT-U = BM
concern)
Unspecified
0 (preferable)
6100
(acceptable)
0 (preferable)
2200
(preferable)
0 (preferable)
10700
(acceptable)
BM2
Moderate
Group I Human
BM2
High Group II
Human
BM1
High Group I
Human
USEPA DfE*
GreenSuite
GreenSuite
(preferred)
(adjusted*)
0 (most concern) to
100% (no concern)
LT-1
High – reproductive and
developmental toxicity
76.1%
71.55%
LT-U
Approved Safer Ingredient
High – Eye irritation
86.75%
78.77%
LT-P1
No Endpoints of Concern
72.32%
69.01%
86.22%
75.49%
73.13%
67.01%
Very High – Eye/Skin
irritation
Moderate to High –
Reproductive and
developmental toxicity
High – Reproductive and
developmental toxicity
Very High – Acute and
chronic aquatic toxicity
0 (preferable)
2200
(preferable)
BM1
High Group I
Human
LT-U
1700
(preferable)
12700
(acceptable)
BM1
High Group I
Human/SVHC
List
LT-1
0 (preferable)
3200
(preferable)
U
Carcinogenicity
LT-U
TBD
77.8%
67.55%
LT-1
High – Developmental
toxicity and persistence
Very High – Acute and
chronic aquatic toxicity
and bioaccumulation
86.57%
79.05%
0 (preferable)
2500
(preferable)
BM1
vBT
53
by American Chemistry Council, Value Chain Outreach Committee, Tools Subcommittee (2014). Publication in preparation.
54
Plan for HESI SCA Technical Committee



IMPROVE the ability of existing tools to identify sustainable alternatives

Identify and demonstrate relevant safety information to inform sustainable
choices

Build consensus on approaches to reduce sources of variation

Build case studies to evaluate and validate tool(s) performance
ENHANCE tools by incorporating advances in the science

Develop principles or tools to support integration of next gen safety data

Demonstrate how exposure data can inform and improve sustainable choices
BUILD OUT to include other LCA-like attributes based on established success with
hazard and exposure
Spring 2015 Workshop in Planning
HESI Scientific Program Manager
Dr. Jennifer Tanir
jtanir@hesiglobal.org
For more information
Animal Alternatives
in Environmental Risk
Assessment
Context for Alternatives Development
Regulatory drivers
Cosmetics Directive
Animal Protection Directive
REACh
Tox21 (regulatory and scientific)
OECD Fish Testing Framework
Scientific drivers
Mechanistic ecotoxicology
AOPs
Endocrine disruption
Modeling mammalian toxicity
Regulatory
Needs
Societal
Needs
Societal drivers
Animal welfare pressure (3R’s approach)
Increased fish use as an alt to higher vertebrates
Reflected in regulatory and scientific worlds
Scientific
Needs
Fish Embryo Test (FET)
• Correlation between FET and acute fish test is better than acute
fish tests between species
• Little difference between zebrafish and fathead minnow with
chemicals tested thus-far
• Already being submitted for registrations / dossiers
• Use of OECD TG 236 data has not translated to acceptance in all
regulatory jurisdictions
• Potential applications beyond acute testing...
Fish Embryo Test (FET) – chronic toxicity
• Work initiated by HESI to explore application /
optimization of the FET to predict chronic toxicity
• Focus on OECD 210 TG (Fish Early Life Stage Test); most
frequently used bioassay to predict chronic toxicity
• Ongoing work to discover, characterize, and annotate FELS
AOPs and prioritize their development (CEFIC LRI, others)
• Ultimate goal is to identify alternative test methods to
predict chronic toxicity in fish
Villeneuve et al. 2014. Environ. Toxicol. Chem. 33: 158-169
Effluent testing
• Effluent testing is the single largest source of use of aquatic
vertebrates globally for a regulatory purpose and is a subject
separate from chemical regulation
•
Initial laboratory research project aimed at developing
approaches to evaluate chronic toxicity of effluents to fish
that extend beyond acute toxicity.
• 2 publications in-press
•
Ongoing work to identify best scientific practices for effluent
testing / assessment
• International workshop on “Concepts, Tools, and
Strategies for Effluent Testing: An International
Workshop” planned for January 2016
Non-testing Approaches: eco-TTC Development
Threshold of Toxicological Concern (TTC)
• An approach for establishing an exposure level for
chemicals, below which no appreciable risk to
human health and/or the environment is expected
• TTC proposes that a de minimis value for toxicity
can be identified for many chemicals, including
those of unknown toxicity
• TTC was originally applied to assess chemicals in
food:
• Flavorings
• Food contact materials
• Pharmaceutical impurities
Environmental TTC Approaches (eco-TTC)
• De Wolf et al. 2005. ET&C 24:479
• Compiled aquatic toxicity data from:
• ECETOC EAT database
• USEPA fathead minnow database
• EU Existing substances database
• Utrecht guppy database
• Binned by Mode of Action (MoA)
• PNECs derived using EU procedures /
Application Factors
• Eco-TTC of 0.1 µg/L for Verhaar MoA 1-3
See also: Gross et al. (2010); Williams et al. (2011) for additional eco-TTC like approaches
Benefits of an eco-TTC
• Maximizes resource use (animals, time, $$)
• Allows screening-level assessment for
chemicals with little or no toxicity data
•
•
•
Groups without a QSAR (or no hope for one)
polymers? UVCBs*?
New chemistries
Difficult to test substances (?)
• Particularly useful for screening assessment
when production is low
• Supports read-across
*Unknown or variable composition, complex reaction products, biological materials
HESI eco-TTC Project
• Led by the HESI Animal Alternatives in Environmental
Risk Assessment Committee
• General strategy to explore this approach in a stepwise manner developed and presented at SETAC,
WC9
• Initial analysis is ongoing, utilizing readily-available
databases (D. deZwart, RIVM) and industry data
when made available
• Strong interest from multiple sectors / groups
• Exploring additional data sources and partners
HESI Sr. Scientific Program Manager
Dr. Michelle Embry
membry@hesiglobal.org
For more information
Genotoxicity Testing
Committee
 Frameworks for integration of
testing results into a riskbased assessment
 Integration and use of
new/emerging technologies
GTTC’s: Improve the scientific basis of the
interpretation of results from genetic toxicology tests
for purposes of more accurate hazard identification
and assessment of human risk.
Clean Sheet Testing Strategy:
Can we do better with a new
approach?
New Approach
•
Accurate

Standard battery?
•
Efficient

Role of MoA?
•
Systems biology-based

•
New technologies
How to define tiers and
need for addn’l testing?

Epigenetics and germ
cells?

Defining risk?
Draft approaches in
summer 2015
Leaders:
Key Questions
Kerry Dearfield (USDA), Mirjam Luijten (RIVM), Bhaskar Gollapudi (Consultant)
PIG-A Assay
-in vivo rodent gene mutation
assay
-faster and less expensive,
relevant results
-HESI publication and research
history
PLAN OF WORK
-SPSF SUBMITTED TO
THE OECD IN 2014
-POTENTIAL TEST
GUIDELINE REVISION
BASED ON DATA TO
BE SUBMITTED BY HESI
COMMITTEE
HESI Scientific Program Manager
Dr. Jennifer Tanir
jtanir@hesiglobal.org
For more information
Developmental and
Reproductive
Toxicology
Committee
Developmental Toxicity Testing:
Value of the 2nd species
Goals & Objectives:


To assess whether testing a 2nd (non-rodent) species provides
added value and to gain a better understanding of
circumstances in which 2nd species does not add value

Inform the decision around the current proposal to revise ICH
S5
•
Workplan:

•
•
•
Customization of US EPA’s
ToxRefDB
Data entry (Microsoft Accessbased; exportable to Excel for
analyses)
Data analyses
•
•
•
Data (non-registered/non-approved
compounds) submitted by sponsors
•
Written & tabulated
summaries (eCTD)
•
Both rat and rabbit EFD
studies
Data blinded by HESI staff
Sponsoring of RIVM post-doc
Provided access to EMA
registered compounds
2nd Species

Preliminary Results:
 Analysis
by maternal systemic exposure:
 For
the majority of compounds (80%), no meaningful
differences in rat and rabbit maternal systemic exposure at
the fLOAEL
 Results
suggest that differences between species based on
maternal systemic exposure may often be due to stochastic
variation rather than being a specific finding.
 Analysis
by severity of effect type:
 In
general, fetal death was more often observed in rabbit,
whereas malformations occur more often in the rat.
 Additional
parameters (maternal toxicity, TK data, strain and
mode of action) did not show any large differences between
species.
2nd Species


Next Steps:
Dec 2014
2015
• Complete two hazard ID
(analyses) manuscripts
• Complete risk assessment
manuscript
• Present results at SOT
Anticipated Impact:

Data & analysis was presented at ICH level.

Database allows future interrogation of additional
questions re species sensitivity (e.g., Human data
exposure vs. animal data are now being overlaid for
marketed products)
backups
January 2015
Diversified Corporate Sponsorship
Why?
Number of Sponsor Organizations
• Emphasis on Recruitment
• Enhanced diversification of
small & large company
participation
• Downsizing and merger of
some larger companies
44
1.5M
49
1.6M
2012
2011
Year
53
57
59?
1.7M
2013
1.7M
2014
1.7-8M?
2015
HESI Staff
Syril Pettit, MEM,
Executive Director
Nancy Doerrer, MS,
Associate Director
Cynthia Nobles,
Branch Administrator
Michelle Embry, PhD
Senior Scientific
Program Manager
Connie Chen, MPH, PhD
Scientific Program
Manager
Stanley Parish, PhD
Scientific Program Manager
Brianna Farr,
Science Program
Associate
Raegan O’Lone, PhD
Senior Scientific
Program Manager
Jennifer Tanir, PhD
Jennifer Pierson, MPH
Scientific Program Manager Scientific Program Manager
Oscar Bermudez
Science Program
Associate
•
http://www.monticello.org/site/research-and-collections/historic-landscape-institute
Science never appears so beautiful as when applied to the uses
of human life. Thomas Jefferson, 1798. Charlottesville, Virginia.