High-Throughput Screening Methods in Toxicity Testing

Explores the benefits and limitations of the latest high-throughput screening methods

With its expert coverage of high-throughput in vitro screening methods for toxicity testing, this book makes it possible for researchers to accelerate and streamline the evaluation and risk assessment of chemicals and drugs for toxicity. Moreover, it enables them to comply with the latest standards set forth by the U.S. National Research Council's "Toxicity Testing in the 21st Century: A Vision and Strategy" and the E.U.'s REACH legislation. Readers will discover a variety of state-of-the-science, high-throughput screening methods presented by a group of leading authorities in toxicology and toxicity testing.

High-Throughput Screening Methods in Toxicity Testing is divided into five parts:

• General aspects, including predicting the toxicity potential of chemicals and drugs via high-throughput bioactivity profiling
• Assessing different cytotoxicity endpoints
• Assessing DNA damage and carcinogenesis
• Assessing reproductive toxicity, cardiotoxicity, and haematotoxicity
• Assessing drug metabolism and receptor-related toxicity

Each chapter describes method principles and includes detailed information about data generation, data analysis, and applications in risk assessment. The authors not only enumerate the advantages of each high-throughput method over comparable conventional methods, but also point out the high-throughput method's limitations and potential pitfalls. In addition, the authors describe current research efforts to make high-throughput toxicity screening even more cost effective and streamlined.

Throughout the book, readers will find plenty of figures and illustrations to help them understand and perform the latest high-throughput toxicity screening methods.

This book is ideal for toxicologists and other researchers who need to implement high-throughput screening methods for toxicity testing in their laboratories as well as for researchers who need to evaluate the data generated by these methods.

PREFACE ix

CONTRIBUTORS xi

PART I GENERAL ASPECTS

1 ToxCast: Predicting Toxicity Potential Through High-Throughput Bioactivity Profiling 3
Keith A. Houck, Ann M. Richard, Richard S. Judson, Matthew T. Martin, David M. Reif, and Imran Shah

2 High-Throughput Toxicity Testing in Drug Development: Aims, Strategies, and Novel Trends 33
Willem G.E.J. Schoonen, Walter M.A. Westerink, Femke M. van de Water, and Horbach G. Jean

3 Incorporating Human Dosimetry and Exposure Information with High-Throughput Screening Data in Chemical Toxicity Assessment 77
Barbara A. Wetmore and Russell S. Thomas

4 The Use of Human Embryonic Stem Cells in High-Throughput Toxicity Assays 97
Xin Huang, Dan-yan Zhu, and Yi-jia Lou

PART II HIGH-THROUGHPUT ASSAYS TO ASSESS DIFFERENT CYTOTOXICITY ENDPOINTS

5 High-Throughput Screening Assays for the Assessment of Cytotoxicity 109
Andrew L. Niles, Richard A. Moravec, Tracy J. Worzella, Nathan J. Evans, and Terry L. Riss

6 High-Throughput Flow Cytometry Analysis of Apoptosis 129
Francesca de Giorgi and Franc¸ois Ichas

7 High Content Imaging-Based Screening for Cellular Toxicity Pathways 143
Bram Herpers and Bob van de Water

8 The Keratinosens Assay: A High-Throughput Screening Assay to Assess Chemical Skin Sensitization 159
Andreas Natsch

9 High-Throughput Screening Assays to Assess Chemical Phototoxicity 177
Satomi Onoue, Yoshiki Seto, and Shizuo Yamada

PART III HIGH-THROUGHPUT ASSAYS TO ASSESS DNA DAMAGE AND CARCINOGENESIS

10 Ames IITM and Ames Liquid Format Mutagenicity Screening Assays 193
Kamala Pant

11 High-Throughput Bacterial Mutagenicity Testing: VitotoxTM Assay 213
Luc Verschaeve

12 Genotoxicity and Carcinogenicity: Regulatory and Novel Test Methods 233
Walter M.A. Westerink, Joe C.R. Stevenson, G. Jean Horbach, Femke M. van de Water, Beppy van de Waart, and Willem G.E.J. Schoonen

13 High-Throughput Genotoxicity Testing: The Greenscreen Assay 271
Jorg Blumel and Nadine Krause

14 High-Throughput Assays to Quantify the Formation of DNA Strand Breaks 285
Marýa Moreno-Villanueva and Alexander Burkle

15 High-Throughput Versions of the Comet Assay 295
Irene Witte and Andre Stang

16 Automated Soft Agar Colony Formation Assay for the High-Throughput Screening of Malignant Cell Transformation 309
Pablo Steinberg

17 High-Throughput Quantification of Morphologically Transformed Foci in Bhas 42 Cells (v-Ha-ras Transfected BALB/c 3T3) Using Spectrophotometry 317
Kiyoshi Sasaki, Ayako Sakai, and Noriho Tanaka

PART IV HIGH-THROUGHPUT ASSAYS TO ASSESS REPRODUCTIVE TOXICITY, CARDIOTOXICITY,
AND HAEMATOTOXICITY

18 ReProGlo: A New Stem-Cell-Based High-Throughput Assay to Predict the Embryotoxic Potential of Chemicals 343
Frederik Uibel and Michael Schwarz

19 Embryonic Stem Cell Test (EST): Molecular Endpoints Toward High-Throughput Analysis of Chemical Embryotoxic Potential 357
Peter T. Theunissen, Esther de Jong, Joshua F. Robinson, and Aldert H. Piersma

20 Zebrafish Development: High-Throughput Test Systems to Assess Developmental Toxicity 371
Stephanie Padilla

21 Single Cell Imaging Cytometry-Based High-Throughput Analysis of Drug-Induced Cardiotoxicity 385
Min Jung Kim and Joon Myong Song

22 High-Throughput Screening Assays to Evaluate the Cardiotoxic Potential of Drugs 403
Carl-Fredrik Mandenius and Thomas Meyer

23 High-Throughput Screening Assays to Evaluate the Hematotoxic Potential of Drugs 421
Caroline Haglund, Rolf Larsson, and Martin Hoglund

PART V HIGH-THROUGHPUT ASSAYS TO ASSESS DRUG METABOLISM AND RECEPTOR-RELATED TOXICITY

24 High-Throughput Enzyme Biocolloid Systems for Drug Metabolism and Genotoxicity Profiling Using LC–MS/MS 433
James F. Rusling and John Schenkman

25 Higher-Throughput Screening Methods to Identify Cytochrome P450 Inhibitors and Inducers: Current Applications and Practice 453
David M. Stresser and George Zhang

26 High-Throughput Yeast-Based Assays to Study Receptor-Mediated Toxicity 479
Johanna Rajasarkka and Marko Virta

27 Evaluating the Peroxisomal Phenotype in High Content Toxicity Profiling 501
Jonathan Z. Sexton and Kevin P. Williams

28 A Panel of Quantitative Calux R Reporter Gene Assays for Reliable High-Throughput Toxicity Screening of Chemicals and Complex Mixtures 519
Bart van der Burg, Sander van der Linden, Hai-yen Man, Roos Winter, LydiaJonker, Barbara van Vugt-Lussenburg, and Abraham Brouwer

29 DR-Calux R : A High-Throughput Screening Assay for the Detection of Dioxin and Dioxin-Like Compounds in Food and Feed 533
Barbara van Vugt-Lussenburg, Harrie T. Besselink, Bart van der Burg, andAbraham Brouwer

INDEX 547

PABLO STEINBERG, PhD, is Full Professor for Food Toxicology and Replacement/Complementary Methods to Animal Testing as well as Director of the Institute for Food Toxicology and Analytical Chemistry at the University of Veterinary Medicine Hannover. Prior to that, Dr. Steinberg was director of the Institute of Nutritional Science at the University of Potsdam. He has also held research positions at the University of Mainz.