Toxicogenomics-Based Cellular Models. Alternatives to Animal Testing for Safety Assessment

331 Pages·2014·12.798 MB·English

by  Jos Kleinjans (Eds.)

Toxicogenomics-Based
Cellular Models
Toxicogenomics-Based
Cellular Models
Alternatives to Animal Testing
for Safety Assessment
Edited by
Jos Kleinjans, PhD
Professor and Chair
Department of Toxicogenomics
Maastricht University, Maastricht, The Netherlands
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10 9 8 7 6 5 4 3 2 1
List of Contributors
Marjam Alloul-Ramdhani
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands
Scott S. Auerbach
Biomolecular Screening Branch, Division of the National Toxicology Program, NIEHS, Research
Triangle Park, North Carolina
Giulia Benedetti
Division of Toxicology, LACDR, Leiden University, Leiden, The Netherlands
Harrie Besselink
BioDetection Systems b.v., Amsterdam, The Netherlands
André Boorsma
The Netherlands Organisation for Applied Scientific Research, Microbiology, and Systems
Biology, Zeist, The Netherlands
Bram Brouwer
BioDetection Systems b.v., Amsterdam, The Netherlands
Ines Chaves
Department of Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands
Maarten Coonen
Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands
Emanuela Corsini
Department of Pharmacological Sciences, University of Milan, Milano, Italy
Erik H.J. Danen
Division of Toxicology, LACDR, Leiden University, Leiden, The Netherlands
Marjo de Graauw
Division of Toxicology, LACDR, Leiden University, Leiden, The Netherlands
Eugin Destici
Department of Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands;
Department of Medicine, School of Medicine, University of California, San Diego, California
Marja Driessen
RIVM—National Institute for Public Health and the Environment, Bilthoven, The Netherlands
xv
xvi List of Contributors
Jennifer Fostel
National Toxicology Program, National Institute of Environmental Health Sciences, Research
Triangle Park, North Carolina
Abdoelwaheb El Ghalbzouri
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands
Sanne A.B. Hermsen
Center for Health Protection Research (GZB), National Institute for Public Health and the
Environment (RIVM), Bilthoven, The Netherlands
Kristina M. Hettne
Department of Medical Informatics, Biosemantics Group, Erasmus University Medical Center,
Rotterdam, The Netherlands
Wouter T.M. Jansen
PricewaterhouseCoopers Advisory N. V., The Hague, The Netherlands
Barae Jomaa
Division of Toxicology, Wageningen University, Wageningen, The Netherlands
Jos Kleinjans
Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands
Jan A. Kors
Department of Medical Informatics, Biosemantics Group, Erasmus University Medical Center,
Rotterdam, The Netherlands
Dinant Kroese
The Netherlands Organisation for Applied Scientific Research, Risk Analysis for Products In
Development, Zeist, The Netherlands
Robert Luebke
Cardiopulmonary and Immunotoxicology Branch, US Environmental Protection Agency,
Research Triangle Park, North Carolina, USA
Karen Mathijs
Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands
Romana Nijman
Department of Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands
Richard S. Paules
Toxicology and Pharmacology Laboratory, Division of Intramural Research, NIEHS, Research
Triangle Park, North Carolina
List of Contributors xvii
Jeroen Pennings
National Institute for Public Health and the Environment (RIVM), Centre for Health Protection,
Bilthoven, The Netherlands
Aldert H. Piersma
Center for Health Protection Research (GZB), National Institute for Public Health and the
Environment (RIVM), Bilthoven, The Netherlands; Department of Toxicogenomics (TGX),
Maastricht University, Maastricht, The Netherlands
Jan Polman
Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands
Leo S. Price
Division of Toxicology, Leiden Amsterdam Center for Drug Research, Leiden University, Leiden,
The Netherlands
Tessa Pronk
National Institute for Public Health and the Environment (RIVM), Center for Health Protection,
Bilthoven, The Netherlands
Sreenivasa Ramaiahgari
Division of Toxicology, Leiden Amsterdam Center for Drug Research, Leiden University, Leiden,
The Netherlands
Erwin L. Roggen
3Rs Management and Consultancy, Kongens Lyngby, Denmark
Peter Schmeits
RIKILT—Institute of Food Safety, Wageningen University and Research Centre, BU Toxicology
and Bioassays, Wageningen, The Netherlands
Jan Hendrik R.H.M. Schretlen
PricewaterhouseCoopers Advisory N. V., The Hague, The Netherlands
Jia Shao
RIKILT—Institute of Food Safety, Wageningen University and Research Centre, BU Toxicology
and Bioassays, Wageningen, The Netherlands
Rob H. Stierum
The Netherlands Organisation for Applied Scientific Research, Microbiology, and Systems
Biology, Zeist, The Netherlands
Laura Suter-Dick
University of Applied Sciences and Art, Northwestern Switzerland (FHNW), School for Life
Sciences, Institute of Chemistry and Bioanalytics, Muttenz, Switzerland
xviii List of Contributors
Ewa Szalowska
RIKILT—Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The
Netherlands
Cornelis P. Tensen
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands
Elisa C.M. Tonk
Department of Toxicogenomics (TGX), Maastricht University, Maastricht, The Netherlands
Geert R. Verheyen
Drug Safety Sciences, Janssen Research and Development, Beerse, Belgium
Oscar L. Volger
RIKILT Institute of Food Safety, Wageningen University and Research Center, Wageningen,
The Netherlands
Annelieke S. de Wit
Department of Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands
Bart van der Burg
BioDetection Systems b.v., Amsterdam, The Netherlands
Joost van Delft
Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands
Freddy van Goethem
Drug Safety Sciences, Janssen Research and Development, Beerse, Belgium
Henk van Loveren
Center for Health Protection Research, National Institute of Public Health and the Environment,
Bilthoven, The Netherlands
Eugene van Someren
The Netherlands Organisation for Applied Scientific Research, Microbiology, and Systems
Biology, Zeist, The Netherlands
Leo van de Ven
RIVM—National Institute for Public Health and the Environment, Bilthoven, The Netherlands
Bob van de Water
Division of Toxicology, LACDR, Leiden University, Leiden, The Netherlands
Gijsbertus T.J. van der Horst
Department of Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands
Louise von Stechow
Division of Toxicology, LACDR, Leiden University, Leiden, The Netherlands
CHAPTER

1.1
Introduction to
Toxicogenomics-Based
Cellular Models
Jos Kleinjans
Department of Toxicogenomics, Maastricht University, Maastricht, the Netherlands
We live in an era where we witness increased needs for alternative models to ultimately replace the
“gold standard” in repeated-dose toxicity testing: the rodent bioassay. In demand are tests that are
more reliably predicting human health risks, are less costly and time consuming, and are preferably
non-animal based, also to meet with ethical concerns on animal welfare. Since the turn of the millen-
nium, so-called ’omics technologies, coming from the endeavor of unraveling the human genome,
have been increasingly applied to these challenges in chemical safety assessment, in an approach
which is generally referred to as “toxicogenomics.” In addition, a wide range of cellular models,
exploiting human cell lines, human primary cells, and human embryonic stem cells, have been put
to the test. While initial results are promising, the upgrading of such cell assays, the absorbing of the
newest ’omics technologies, and in particular the managing of the tsunami of ’omics data, still pose a
major challenge to the research community of toxicogenomics.
Multiple user groups have high expectations of such toxicogenomics-based approaches towards
developing novel test systems for chemical safety assessment. However, a thorough overview that
will simultaneously introduce toxicogenomics to a wider readership is still lacking.
The goal of this book is to describe the state of the art in developing toxicogenomics-based cel-
lular models for chemical-induced carcinogenicity, immunotoxicity, and reproduction toxicity, all
important endpoints of toxicity, the evaluation of which to date costs large numbers of animal lives.
Also, where ’omics technologies tend to generate “big data” requiring extensive bioinformatics and
biostatistics efforts for actually retrieving toxicologically meaningful results, the field of toxicoinfor-
matics will be thoroughly introduced. The book will also address how to validate toxicogenomics-
based alternative test models, and will provide an outlook to societal and economic implementation
of these novel assays.
3
Toxicogenomics-Based Cellular Models.
© 22001144 Elsevier Inc. All rights reserved.
4 CHAPTER 1.1 Introduction to Toxicogenomics-Based Cellular Models
1.1.1 The demands for alternatives to current animal test models for
chemical safety
The current default model for assessing repeated-dose toxicity of novel or existent chemicals is the
rodent bioassay involving mice or rats. Sometimes, however, also guinea pigs and rabbits, and on rare
occasions dogs or monkeys, are used. Options here are the 28-day oral toxicity test, the 28-day der-
mal toxicity test, and the 28-day inhalation toxicity test. For sub-chronic toxicity testing, 90-day oral,
dermal, or inhalation toxicity studies are available. Lastly, chronic rodent assays have been put in
place, such as the 2-year treatment protocol for carcinogenicity testing. All protocols involve the daily
administration of the compound of interest. These assays aim to quantitatively analyze whether and to
what extent toxicity, a persistent or progressively deteriorating dysfunction of cells, organs, or multi-
ple organ systems, is present upon repeated administration to the animal of the chemical under inves-
tigation. Repeated-dose testing in vivo enables evaluating particular molecular and histopathological
endpoints of toxicity in organs, but also provides information on perturbations of more complex (e.g.
hormonal, immunological, neurological) systems. Focus is on establishing dose–response relation-
ships, from which a NOAEL (no observable adverse effect level) is derived that forms the basis for
setting safety standards for human health in relation to daily lifetime exposure to the chemical. For
formalized safety testing, international regulatory authorities such as the Organization of Economic
Cooperation and Development (OECD) have developed a series of dedicated guidelines. It should
be noted that toxicity testing protocols may differ to some extent, depending on the domain of ulti-
mate application—for example, pharmaceuticals, cosmetics, or food. Current estimates calculate that
approximately 14% of all animals annually tested within the EU for the purpose of chemical safety
assessment are used in sub-chronic and chronic safety assessments, e.g. repeated-dose toxicity.
Obviously, the underlying assumption for using animals in safety testing is that results can be
extrapolated to humans relatively reliably, because of resemblances at the molecular and physiologi-
cal levels: basically, we are all mammals. It may be convincingly argued that the repeated-dose ani-
mal test has probably prevented mankind in the past from major chemical disasters. By contrast, it is
worthwhile to note that your average Shakespearian king, under constant threat of being poisoned,
would not have trusted an animal for his private chemical safety testing but would have required a
serf for pre-tasting his food. In more civilized terms, over the last decade or so, we have learned to
ask critical questions concerning the actual relevance of the rodent bioassay for assessing chemical
safety to human health. A short overview follows.
Over the last decade, the pharmaceutical industry has suffered from high attrition rates of novel can-
didate drugs, in particular because of adverse findings in the last research and development stages—for
example, during clinical trials. These related to disappointing efficacies, or inadequate adsorption, dis-
tribution, metabolism and excretion (ADME) properties, but in 30–40% of cases also to overt toxicity,
in particular for the liver and the heart, despite the fact that animal tests had reported these novel com-
pounds to be safe [1]. Unexpected toxicity in humans may even occur after market introduction. Each
year, about two million patients in the United States experience a serious adverse drug reaction when
using marketed drugs, resulting in 100,000 deaths, thus representing the fourth leading cause of death
[2]. Similar percentages have been estimated for other Western countries such as The Netherlands [3].
Here is a case where the “gold standard” animal model for chemical safety clearly lacks sufficient sen-
sitivity. Failure in the last phases of drug development obviously is to the disadvantage of patients, but
also, in view of the extreme costs of developing new drugs, involves huge economic losses [4].
1.1.2 The toxicogenomics approach 5
Simultaneously, other examples demonstrate that animal models for repeated-dose toxicity may
also over-report human health risks: the US Physicians' Desk Reference has reported that out of 241
pharmaceutical agents used for chronic treatment, 101 agents were demonstrated to be carcinogenic
to rodents. However, epidemiological studies among chronically treated patients as reviewed by the
World Health Organization (WHO) International Agency on Research on Cancer have identified only
19 pharmaceuticals, mostly intended for anticancer treatment or hormone therapy, to be actually car-
cinogenic to man.
This apparent lack of sufficient specificity and sensitivity of the rodent bioassay for repeated-dose
toxicity is underscored by a report indicating that only 43% of toxic effects of pharmaceuticals in
humans were correctly predicted by tests in rodents [5].
These examples demonstrate that better tests for predicting human drug safety are in demand.
Next, there also are ethical concerns with animal toxicity testing. In general, the way animal wel-
fare is considered has often be claimed to represent a sign of civilization. In the words of the twen-
tieth century Indian civil rights activist and political leader Mahatma Gandhi, “The greatness of a
nation and its moral progress can be judged by the way its animals are treated.” Immanuel Kant, the
great eighteenth century German moral philosopher, had already stated that “We can judge the heart
of a man by his treatment of animals.” With regard to animal experimentation, also implying the use
of animal tests for chemical safety evaluations, these concepts were successfully adopted by William
Russell and Rex Burch in their 1959 book The Principles of Humane Experimental Technique, in
which they presented the 3R principle, referring to replacement, refinement, and reduction of ani-
mal testing. Since then, the 3R principle has also found political recognition, for instance within the
EU where the Protocol on Protection and Welfare of Animals annexed to the European Community
(EC) Treaty aims at ensuring improved protection and respect for the welfare of animals as sentient
beings. It is stated that in formulating and implementing the Community’s policies, the Community
and the member states shall pay full regard to the welfare requirements of animals. All industry sec-
tors, including pharmaceuticals, chemicals, cosmetics, agrochemicals, and foods manufacturers, are
consequently obliged to apply available methods to replace, reduce, and refine animal use (three Rs)
in safety and efficacy evaluations under the existing animal protection legislation (Directive 86/609/
EEC) [6]. The most prominent political action within the EU in this context, concerning safety testing
of cosmetic ingredients, undoubtedly is the 7th Amendment (Directive 2003/15/EC) to the Cosmetics
Directive, which requires the full replacement of animals in safety testing, and which sets a timetable
for the availability of alternative testing methods for assessing safety of cosmetics ingredients and
products, with a deadline of 2013.
All in all, we now see the need for better, reliable predictive tests for assessing chemical safety for
human health, which come less costly and less time consuming, and preferably are no longer animal
based.
1.1.2 The toxicogenomics approach
With the advent of genomics technologies to the domain of toxicology, hopes have been set high
that the so-called toxicogenomics approach may actually deliver the desired alternative test systems
to the current animal models for chemical safety. This is, for instance, expressed in the 2006 EU
Regulation on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH),