NorCal Society of Toxicology Fall Symposium
Advances in Predictive and Investigative Toxicology
Thursday, November 4th, 2010
Genentech, Building 42, Conf Rooms D/E
475 East Grand Avenue, South San Francisco, CA 94080
Registration and Continental Breakfast:
9:20 – 9: 40
8:00-9:20
Welcome and Chapter Business Updates: Tao Wang, President for NorCal SOT
9:40 – 10:20 Multi-Hit Models of Neurotoxicity: Implications for Understanding Disease and Improving
Risk Assessment: Deborah Cory-Slechta, Professor, University of Rochester School of
Medicine and Dentistry
10:20 – 11:00 Advances in Preclinical Immunotoxicology Safety Assessment: Michael P. Holsapple,
Executive Director, ILSI Health and Environmental Sciences Institute
11:00 – 11:40 Applying NMR- and LC/MS-Based Metabolite Profiling to Predictive and Investigative
Toxicology: Lois Lehman-McKeeman, Distinguished Research Fellow, Discovery
Toxicology, Bristol-Myers Squibb
Lunch (Lunch with an Expert program):
11:40 – 1:00
1:00 – 1:40
From Descriptive to Predictive Environmental Toxicology at the U.S. EPA: David Dix,
Acting Deputy Director, National Center for Computational Toxicology, US EPA
1:40 – 2:20
Investigative and Predictive Approaches to Drug-Induced Steroid Hormone Perturbation:
Jeff Lawrence, Director, Investigative Toxicology, Amgen
2:20 – 3:00
BREAK: vendors can move to 42C; poster setup in 42C
3:00- 3:40
Exploratory Cardiovascular Safety Pharmacology in the Lead-Optimization of Novel Small
Molecule Kinase Inhibitors: Michael Flagella, Scientist, Investigative Safety Assessment,
Genentech
3:40 – 4:20
Pharmacokinetic Drivers of Toxicity for Small Molecules: Evaluating Plasma-Tissue
Concentration Relationships: Dolo Diaz, Scientist, Investigative Safety Assessment,
Genentech
Wine and Cheese/ poster viewing:
4:30-5:30
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Registration
Pre-registration will facilitate planning for security and parking purposes. Registration is
$25 for NorCal SOT members, $35 for nonmembers and FREE for students/post-docs.
Additional $5 will be applied to onsite registration. Students are encouraged to bring posters
documenting their research and will receive $50 for doing so. Students may have their posters
printed by contacting Steve Dizio at sdizio@dtsc.ca.gov.
Sponsors
This meeting is generously sponsored by the following:
BioReliance
Charles River
Genentech
Covance
MPI
SRI
ChanTest Corp
Toxicon
Pacific BioLabs
Huntingdon Life Sciences
ITR Lab
Zenas Technology
BMG Lab Tech
Battelle
Abstracts and Biographical sketches
Deborah Cory-Slechta, Professor, University of Rochester School of Medicine and Dentistry
Multi-Hit Models of Neurotoxicity: Implications for Understanding Disease and Improving Risk
Assessment
It is increasingly recognized that most human diseases and disorders are multi-factorial in etiology. Yet both
experimental models and epidemiological studies evaluating the potential contributions of environmental or
occupational chemical exposures to these diseases generally focus on the impact of a single chemical exposure
in isolation from other risk factors. This includes interactions with other chemical exposures, as is more
consistent with the human environment. This talk will present the results from studies of multi-hit models of
pesticide exposures in conjunction with other risk factors for the Parkinson’s disease phenotype, and for
cognitive deficits in children in response to elevated lead exposure combined with prenatal stress, cooccurring risk factors for low socioeconomic status communities. Based on such approaches, it will suggest
strategies for improving animal models and epidemiological studies of neurotoxicity to advance our
understanding of diseases and disorders, and to move forward cumulative risk assessment by using common
adverse outcomes to define those chemical exposures and other risk factors that should be evaluated in
combination.
Bio: Dr. Deborah Cory-Slechta became a faculty member at the University of Rochester Medical School
(URMC) in 1982 where she rose from Assistant Professor to Professor. She became Chair of its Department
of Environmental Medicine and Director of the NIEHS Environmental Health Sciences Center in 1998, and
served as Dean for Research from 2000-2002.From 2003-2007, she served as Director of the Environmental
and Occupational Health Sciences Institute (EOHSI) and Chair of the Department of Environmental and
Community Medicine at the UMDNJ-Robert Wood Johnson Medical School in New Jersey, before returning
to URMC as Professor in Environmental Medicine and Pediatrics. Dr. Cory-Slechta has served on national
review and advisory panels of the National Institutes of Health, the National Institute of Environmental Health
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Sciences, the Food and Show Desktop.scf Drug Administration, the National Center for Toxicological
Research, the Environmental Protection Agency, the National Academy of Sciences, the Institute of Medicine,
and the Agency for Toxic Substances and Disease Registry, Centers for Disease Control. She currently serves
on the Science Advisory Board of the US EPA and on the Advisory Committee for Childhood Lead Poisoning
Prevention of the CDC. In addition, Dr. Cory-Slechta has served on the editorial boards of the journals
Neurotoxicology, Toxicology, Toxicological Sciences, Fundamental and Applied Toxicology,
Neurotoxicology and Teratology, and American Journal of Mental Retardation. She has held the elected
positions of President of the Neurotoxicology Specialty Section of the Society of Toxicology, President of the
Behavioral Toxicology Society, and been named a Fellow of the American Psychological Association. Her
research has focused largely on the relationships between brain neurotransmitter systems and behavior, and
how such relationships are altered by exposures to neurotoxicants, particularly the role played by
environmental neurotoxicant exposures in developmental disabilities and neurodegenerative diseases. These
research efforts have resulted in over 120 papers and book chapters to date.
Michael P. Holsapple, Executive Director, ILSI Health and Environmental Sciences Institute
Advances in Preclinical Immunotoxicology Safety Assessment
This presentation will capitalize on the nearly 30 years of experience by the speaker as an
immunotoxicologist. The advances in preclinical immunotoxicology safety assessment will be addressed
from two important perspectives. The first perspective will be to consider the current state-of-the-science of
immunotoxicity testing, as it relates to regulatory guidelines for immunotoxicology for both drugs and
chemicals. The second perspective will be to highlight some of the contributions and current activities by the
Immunotoxicology Technical Committee (ITC), one of the oldest and most productive components of the
scientific portfolio within the ILSI Health and Environmental Sciences Institute (HESI).
Bio: Dr. Holsapple is a toxicologist with over 30 years of experience. He received his graduate training in
Pharmacology and Toxicology from Purdue University, having earned an M.S. in 1978 and a Ph.D. in 1981.
From 1983–1994, he served on the faculty at the Medical College of Virginia/Virginia Commonwealth
University in Richmond, VA. During his academic career, he served as the advisor for 8 Ph.D. and M.S.
candidates, and as a member of the doctoral dissertation committees for 21 other students. From 1994-2002,
Dr. Holsapple worked in the Toxicology, Environmental Research and Consulting Laboratories at the Dow
Chemical Company in Midland, MI. During his industry career, his responsibilities included leading both the
Immunotoxicology and the Respiratory Toxicology groups. Dr. Holsapple is currently the Executive Director
of the Health and Environmental Sciences Institute (HESI), the global branch of the International Life
Sciences Institute (ILSI). During his time with HESI, Dr. Holsapple has facilitated the organization’s
emergence as a recognized global leader in advancing the state-of-the-science of safety and risk assessment.
Dr. Holsapple has published over one hundred and fifty manuscripts and chapters. In recognition of his
contributions to toxicology, Dr. Holsapple received the Society of Toxicology (SOT) Achievement Award in
1992. In recognition for his contributions to immunotoxicology, he received the Vos Award – Career
Achievement in Immunotoxicology in 2009. Dr. Holsapple served on Council for the SOT from 2005-2007,
and on Council for the American College of Toxicology (ACT) from 2003-2006. He was elected a ‘fellow’ in
the Academy of Toxicological Sciences (A.T.S.) in 2006. Dr. Holsapple was elected into the Presidential
track of the SOT in 2008, and is currently the President.
Lois Lehman-McKeeman, Ph.D.Distinguished Research Fellow, Discovery Toxicology, Bristol-Myers
Squibb Co., Princeton, NJ
Applying NMR- and LC/MS-Based Metabolite Profiling to Predictive and Investigative Toxicology
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Metabolomic profiles can provide important leads for identifying key events underlying toxicologic responses.
Furthermore, as a likely phenotypic consequence of altered gene expression, metabolomic changes are
important complementary data for determining the systemic relevance or significance of induction or
repression of specific gene pathways. In this presentation, examples of how NMR- and LC/MS-based
metabolite profiling have aided the identification of potential biomarkers and/or mechanisms of toxicity will
be discussed. In particular, urinary profiles facilitated the discovery of methylhistidine metabolites as
potential new biomarkers for assessing skeletal muscle toxicity. Evaluation of urine, serum and tissue lipid
profiles have also afforded new perspectives on biochemical changes predictive of and mechanistically
relevant for evaluating drug-induced phospholipidosis. There is potential for metabolomic investigations to
identify non-invasive tools that translate from preclinical to clinical assessments and thereby facilitate human
safety evaluation.
Bio: Dr. Lois Lehman-McKeeman is currently a Distinguished Research Fellow in Discovery Toxicology at
the Bristol-Myers Squibb Company in Princeton, NJ. She received a BS degree in Toxicology from the
University of the Sciences in Philadelphia and holds a Ph.D. in Toxicology from the University of Kansas
Medical Center. She was employed in the Human and Environmental Safety Division of the Procter and
Gamble Company for 15 years prior to joining Bristol Myers Squibb in 2001. Lois has active research
interests and programs broadly in biochemical mechanisms of toxicity and non-genotoxic mechanisms of
carcinogenesis. She is also working to develop and apply metabonomic and transcriptomic technologies to
mechanistic toxicology. She has published extensively in these fields. She has been active professionally in
the Society of Toxicology (SOT) serving on numerous SOT committees, and she held elective office in the
SOT as Councilor (2000-2002) and the SOT Awards Committee (2008-2010). In 2003 she was appointed
Editor of Toxicological Sciences, a position she currently holds, and she serves on a number of other editorial
boards. She was a member of the Human Subjects Review Advisory Board of the USEPA (2006-2010) and
she has or is presently sitting on a variety of national and international advisory committees for EPA, NIH and
IARC. She is a Trustee on the Board of Directors of the International Life Sciences Human and
Environmental Safety Institute (ILSI-HESI). She was elected as a Fellow of the American Association for the
Advancement of Science in 2008 and has been a fellow in the Academy of Toxicological Sciences since 2000.
She was the recipient of the Robert Scala Award in Toxicology for research excellence in an industrial
laboratory (1994), the Society of Toxicology Achievement Award (2003) and the George H. Scott Award for
scientific excellence from the Toxicology Forum (2006).
David Dix, PhD, Acting Deputy Director, National Center for Computational Toxicology, US EPA
High Throughput Screening For Hazard And Risk Of Environmental Contaminants
High throughput toxicity testing provides detailed mechanistic information on the concentration response of
environmental contaminants in numerous potential toxicity pathways. High throughput screening (HTS) has
several key advantages: (1) expense orders of magnitude less than animal testing; (2) direct study of human
gene, protein and cell targets is possible; and (3) hundreds to thousands of contaminants can be studied
simultaneously. Quantitative HTS hazard assessment can thus identify potential mechanisms and pathways by
which a contaminant can lead to specific adverse outcomes. EPA’s ToxCast project is evaluating the use of
HTS for understanding the types of molecular and pathway perturbations caused by environmental chemicals,
and building initial predictive models of in vivo toxicity for prioritization and hazard assessment. To date we
have tested 309 pesticide active and industrial chemicals in 467 assays across 9 HTS technologies. These
include cell-free assays, as well as cell-based assays in a variety of human and rodent primary cells and cell
lines, Both individual assays and composite assays for effects on genes and pathways demonstrated a broad
spectrum of chemical activity at the molecular and pathway levels. Many expected interactions were seen in
the data, including endocrine and xenobiotic metabolism enzyme activity.
Chemicals show widely varying promiscuity across pathways, from no activity to activity in dozens of
pathways. This diversity of behavior is seen even within well-defined chemical classes. This approach
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promises to provide meaningful data on thousands of untested environmental chemicals, and to guide more
intelligent, targeted testing of environmental contaminants in the future. This abstract may not necessarily
reflect Agency policy.
Bio: Dr. David J. Dix is a Research Biologist and Acting Deputy Director of the U.S. Environmental
Protection Agency’s National Center for Computational Toxicology (NCCT), at Research Triangle Park, NC,
USA. NCCT develops high-throughput decision support tools for screening and assessing chemical exposure,
hazard and risk. For example, ToxCastTM utilizes high-throughput screening technologies to derive
bioactivity profiles that predict toxicity, characterize toxicity pathways, and prioritize environmental
chemicals for further testing. Dr. Dix is an adjunct Associate Professor in the Department of Environmental
Sciences and Engineering at the University of North Carolina at Chapel Hill. He earned his undergraduate
degree in Biological Sciences from the University of Illinois at Chicago, a Ph.D. in Physiology from Rush
University in Chicago, and completed postdoctoral training at the U.S.National Institute of Environmental
Health Sciences. He has published over 90 articles, reviews and book chapters, serves on several Editorial
Boards, and has given numerous national and international presentations on his research.
Jeffrey W. Lawrence, Ph.D, Director, Biochemical Toxicology and Biomarkers, Amgen, Inc.
Investigative and predictive approaches to drug-induced steroid hormone perturbation
During development, a candidate molecule demonstrated male reproductive toxicity suggesting testosterone
hormone disruption. In vitro models and an array of biochemical methods were used to determine the site of
perturbation. The mechanism was found to only affect the signaling portion of testosterone synthesis. An in
vivo metabolomics study was conducted to bridge the putative mechanism to an animal model. These results
from this study confirmed the mechanism and demonstrated a distinct profile from an agent with a different
hormone disruption mechanism. Consequently, an in vitro assay was used to survey other molecules to
prioritize them for development.
Bio: Jeff Lawrence is the Director of Biochemical Toxicology and Biomarkers in the Department of
Investigative Toxicology at Amgen in Thousand Oaks, CA. Jeff Lawrence holds a B.Sc. degree in
Toxicology from the Philadelphia College of Pharmacy and Science, and a Ph.D in Pharmacology from the
University of Florida. During his dissertation, Jeff studied DNA toposiomerases and mtDNA depletion
mechanisms under the direction of Dr. Tom Rowe. Jeff completed his postdoctoral training at Eli Lilly and
Co, where he worked with Dr. Pat Eacho in the Hepatotoxicity laboratory in the Biochemical Toxicology
group and later the Cardiovascular Discovery Research group. Following his postdoctoral training, Jeff
worked in the Department of Safety Assessment at Merck and Co., Inc. for 10 years. During this time, he was
involved in toxicokinetic analysis for various drug development programs and actively led and participated in
Investigative Toxicology research. Jeff Joined Amgen, Inc. in 1996 as Director, Biochemical Toxicology and
Safety Biomarkers. His investigative efforts use biochemical, cellular, and molecular based approaches using
in vitro and in vivo systems to solve program issues jeopardizing continued development of drug candidates.
Michael Flagella, Ph.D. Investigative Safety Assessment, Genentech
Exploratory cardiovascular safety pharmacology in the lead-optimization of novel small molecule
kinase inhibitors
Early prediction of potential cardiac liabilities and selection of molecules without these liabilities in the
research phase will enable an overall decrease in compound attrition at later stages of drug development.
During the early lead optimization phase of novel small molecule kinase inhibitors, we observed acute, lethal
toxicities in some, but not all, structurally-similar molecules at subefficacious exposures. These toxicities were
consistent with functional changes to neuromuscular or cardiovascular systems, and were not attributable to
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inhibition of the intended target kinase. Overlapping patterns of significant off-target inhibition (muscarinic,
GABAa GPCRs and acetylcholinesterase) provided insight into the observed toxicities. Our approach
integrated cardiac safety pharmacology testing (e.g. ex vivo Langendorff preparations and rodent telemetry
systems) into medicinal chemistry. We iteratively utilized in vitro receptor binding and functional assays with
in vivo counter-screens to rank-order compounds with the lowest potential for cardiac safety liabilities.
Efficacious, free-drug exposures were used to guide safety pharmacology experiments and to identify a lead
candidate based on the best possible safety characteristics and margin. Studies in telemeterized monkeys
identified minor, transient blood pressure decreases at therapeutic exposures, thus validating our approach.
Coupled with repeat-dose safety studies, a lead candidate compound with minimal cardiac effects was
nominated for IND-enabling studies.
Bio: Mike is a scientist in the Investigative Safety Assessment group at Genentech, and provides proactive
discovery toxicology team representation to enable toxicology-related decision-making during discovery, lead
optimization and early development.
Mike received his Ph.D. in Molecular Genetics from the University Of Cincinnati College Of Medicine and
his B.S. in Recombinant Gene Technology from the State University of New York College at Fredonia. Prior
to arriving at Genentech in 2008, he was a cell biologist at Novabay Pharmaceuticals, a gene-expression assay
development scientist at Panomics (now Affymetrix), and a post-doctoral fellow in the Systems Biology group
at Eli Lilly and Co.
Dolo Diaz, Ph.D. Investigative Safety Assessment, Genentech
Pharmacokinetic Drivers of Toxicity for Small Molecules: Evaluating Plasma-Tissue Concentration
Relationships
Plasma drug exposures are routinely used in the interpretation of animal toxicity data. In reality, target tissue
exposures can be much higher than plasma exposures, which could have implications in the understanding of
in vivo toxicities and in the determination of in vivo-in vitro correlations. Examples will be presented in
which in vitro-in vivo correlations and differences in species sensitivity can be better understood by
considering target organ exposures. Systematic analysis of tissue drug levels for a group of structurally
diverse small molecules reveals large differences in tissue distribution; in addition, tissue exposure levels
show improved correlation with toxicity compared to plasma exposure levels. Analysis of pharmacokinetic
parameters for these compounds indicates that tissue distribution can be roughly predicted by taking into
consideration the volume of distribution (Vss) and the clearance (CLp) of a particular molecule. These
findings suggest that consideration of these pharmacokinetic parameters could potentially help provide more
relevant and translatable safety information.
Bio: Dolo has been a Scientist in Investigative Safety Assessment at Genentech for the past 3.5 years, where
she focuses on discovery and investigative toxicology for small molecules. Before joining Genentech, she
was a group leader for in vitro toxicology at Cerep (4 years). Areas of expertise include lead optimization,
hepatotoxicity, genotoxicity, investigative toxicology. She has a BS in Pharmacy, a PhD in Toxicology from
the University of Washington (developmental toxicology, metal toxicology) and postdoctoral training at the
Fred Hutchinson Cancer Research Center (cancer biology); she is a Diplomat of the American Board of
Toxicology.
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