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Molecular and Cellular Toxicology: An Introduction

Molecular and Cellular Toxicology: An Introduction

Lesley Stanley

ISBN: 978-1-119-95207-7

Jun 2014

434 pages

In Stock

$154.95

Description

Toxicology is the study of the adverse effects of chemical, physical, or biological agents on people, animals, and the environment. Toxicologists are trained to investigate, interpret, and communicate the nature of those effects. 

Over the last ten years the subject of toxicology has changed dramatically, moving from a discipline which was once firmly wedded to traditional methods to one which is keen to embrace the innovative techniques emerging from the developing fields of cell culture and molecular biology. There is an acute need for this to be reflected in a paradigm shift which takes advantage of the opportunities offered by modern developments in the life sciences, including new in vitro and in silico approaches, alternative whole organism (non-mammalian) models and the exploitation of ‘omics methods, high throughput screening (HTS) techniques and molecular imaging technologies.

This concise, accessible introduction to the field includes the very latest concepts and methodologies. It provides MSc, PhD and final year undergraduate students in pharmacy, biomedical and life sciences, as well as individuals starting out in the cosmetics, consumer products, pharmaceutical and testing industries, with everything they need to know to get to grips with the fast moving field of toxicology and the current approaches used in the risk assessment of drugs and chemicals.

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Foreword xiii

Preface xv

Acknowledgements xvii

Abbreviations xix

About the Companion Website xxiii

1 Background to Molecular and Cellular Toxicology 1

1.1 What do we mean by molecular and cellular toxicology? 1

1.2 Tissues and their maintenance 2

1.2.1 Stem cells 3

1.3 Tissue damage 4

1.3.1 Consequences of tissue injury 4

1.3.2 Reversible changes in cells and tissues 6

1.3.3 Irreversible changes in cells and tissues 7

1.4 Tissue responses to injury 7

1.4.1 Oxidative stress 7

1.4.2 Necrosis and apoptosis 10

1.4.3 Neoplasia 13

1.4.4 The initiation–promotion paradigm 13

1.5 Key concepts in toxicology 23

1.5.1 Risk and hazard 23

1.5.2 Variability and uncertainty 25

1.5.3 Threshold and non-threshold dose responses 26

1.5.4 The regulatory context 28

1.5.5 Limitations of whole animal studies 29

1.5.6 Use of human tissues in toxicology 31

1.6 Summing up 33

Self-assessment questions 33

Background Reading 34

References 34

2 Individual Susceptibility to Toxic Chemicals 37

2.1 Introduction 37

2.2 Toxicogenetics and toxicogenomics 38

2.3 Genotyping and phenotyping 39

2.3.1 Genotyping 40

2.3.2 Phenotyping 43

2.3.3 Correlating genotype and phenotype 44

2.4 Polymorphic xenobiotic metabolism 45

2.4.1 Polymorphic xenobiotic metabolising enzymes 49

2.4.2 The role of xenobiotic metabolising polymorphisms in susceptibility to toxic agents 50

2.5 Study numbers and effect size 60

2.6 Recent developments 62

2.6.1 Genome-wide association studies 62

2.6.2 Collaborative programmes 64

2.7 The UK Biobank 69

2.8 Conclusions 71

Self-assessment questions 72

Background Reading 72

References 73

3 ‘Omics Techniques 79

3.1 ‘Omics and bioinformatics 79

3.2 Transcriptomics 80

3.2.1 Methodology 80

3.2.2 Proof of principle 89

3.2.3 Hepatotoxicity 91

3.2.4 Extrahepatic toxicity 96

3.3 Proteomics 97

3.3.1 Methodology 98

3.4 Metabolomics/metabonomics 101

3.4.1 MS-based metabolomics 102

3.4.2 NMR-based metabolomics 106

3.5 Integrating different types of ‘omics data 107

3.5.1 ‘Omics in drug discovery 108

3.5.2 ‘Omics profiles as biomarkers of toxicity 109

3.6 Remaining issues with ‘omics approaches 111

3.7 Conclusions 112

Self-assessment questions 113

Background Reading 113

References 113

4 In Vitro Methods for Predicting In Vivo Toxicity 117

4.1 In vitro toxicology 117

4.2 Tissue culture 117

4.2.1 Primary cell cultures 122

4.2.2 Established cell lines 125

4.3 Acute toxicity in vitro 127

4.3.1 Cytotoxicity testing 127

4.3.2 Choice of cell line 129

4.3.3 Liver 131

4.3.4 Skin 133

4.3.5 Eye 141

4.4 Repeated dose toxicity 144

4.5 Reproductive toxicity 147

4.6 Stem cell-derived systems 149

4.7 Conclusions 151

Self-assessment questions 151

Background Reading 152

References 152

5 In Vitro Methods for Absorption, Distribution, Metabolism and Excretion 159

5.1 Why study ADME in vitro? 159

5.2 Absorption 160

5.2.1 Dermal penetration 160

5.2.2 Gastrointestinal absorption 164

5.3 Distribution 171

5.3.1 Protein binding 172

5.3.2 Blood-brain barrier 172

5.3.3 Other protective barriers 176

5.4 Metabolism 176

5.4.1 Skin 177

5.4.2 Gastrointestinal tract 179

5.4.3 Liver 179

5.5 Excretion 189

5.5.1 Biliary excretion 190

5.5.2 Renal clearance 191

5.6 Conclusions 191

Self-assessment questions 192

References 192

6 In Silico Methods and Structure–Activity Relationships 199

6.1 Why in silico? 199

6.2 Predicting the ADME characteristics of xenobiotics 200

6.2.1 Absorption 200

6.2.2 Distribution 206

6.2.3 Metabolism 207

6.2.4 Excretion 212

6.3 Physiologically based biokinetic modelling 212

6.4 Toxicity 221

6.4.1 Exposure modelling 222

6.4.2 Prediction of toxicity 223

6.5 Conclusions 233

Self-assessment questions 235

References 235

7 Transgenic Animal Models for ADME and Systemic Toxicity 241

7.1 Transgenic models and their use in toxicology 241

7.2 ADME models 242

7.2.1 Nuclear receptor models 244

7.2.2 Xenobiotic metabolism models 251

7.2.3 Drug transporter models 259

7.3 Reporter models 264

7.3.1 LacZ-based models 264

7.3.2 Green fluorescent protein-based models 267

7.3.3 Luciferase-based models 268

7.3.4 Evaluation 273

7.4 Conclusions 273

Self-assessment questions 274

Background Reading 274

References 274

8 Genotoxicity and its Measurement 281

8.1 Genotoxicity testing 281

8.2 Core in vitro tests 282

8.2.1 The Ames test 282

8.2.2 In vitro gene mutation tests using mammalian cells 283

8.2.3 The in vitro chromosome aberration test 284

8.2.4 The in vitro micronucleus assay 287

8.3 Assessment of genotoxicity for regulatory purposes 291

8.4 Novel in vitro methods 292

8.4.1 GreenScreen HC 292

8.4.2 The Reconstructed Skin MicroNucleus assay 293

8.5 Novel in vivo assays for gene mutations 294

8.5.1 The Pig-A assay 294

8.5.2 In vivo assays using transgenic mouse models 295

8.6 DNA damage and its repair 300

8.6.1 DNA damage 300

8.6.2 DNA repair 305

8.7 Thresholds 308

8.8 Conclusions 310

Self-assessment questions 310

References 311

9 Oncogenes and the Identification of Human Carcinogens 317

9.1 Introduction 317

9.2 Identification of human carcinogens 317

9.2.1 The lifetime carcinogenicity bioassay 317

9.2.2 The National Toxicology Program 2-year bioassay 318

9.3 Genetic changes in cancer 321

9.3.1 Methods for detecting activated oncogenes 324

9.3.2 In vitro transformation assays 324

9.3.3 Ras oncogene activation during tumour development 326

9.3.4 Non-ras oncogenes 328

9.3.5 Evaluation 329

9.4 Non-genotoxic carcinogenesis 329

9.4.1 Non-receptor-mediated mechanisms 330

9.4.2 Receptor-mediated mechanisms 331

9.4.3 When is a genotoxic carcinogen not a genotoxic carcinogen? 333

9.5 Transgenic models for short-term carcinogenicity bioassays 335

9.5.1 RasH2 335

9.5.2 Tg.AC 337

9.5.3 p53 models 338

9.5.4 XPC−¨M−, XPA−¨M− and XPA−¨M−/p53+¨M− null mouse models 340

9.5.5 Comparative evaluation of models 340

9.5.6 Regulatory status 341

9.5.7 Limitations of the assays 343

9.5.8 Evaluation 344

9.6 Conclusions 345

Self-assessment questions 346

References 346

10 Emerging Techniques 351

10.1 What’s next? 351

10.2 Novel model organisms 351

10.2.1 The zebrafish 352

10.2.2 Evaluation 358

10.3 Less invasive methods 359

10.3.1 Use of biomarkers 359

10.3.2 Liver 359

10.3.3 Kidney 367

10.3.4 Circulating mRNA biomarkers 371

10.3.5 Evaluation 373

10.4 The systems biology approach 373

10.4.1 Systems biology in toxicology 376

10.5 Collaborative programmes 381

10.5.1 Europe 381

10.5.2 USA 383

10.5.3 Evaluation 384

10.6 Final word 385

Self-assessment questions 385

References 385

Index 391

“Overall, we consider that this book is a useful summary of current and emerging techniques in molecular toxicology.”  (BTS Newsletter, 1 March 2015)