SCOPE 52IPCS JOINT ACTIVITY 19SGOMSEC 8

Methods to AssessDNA Damageand Repair:Interspecies ComparisonsEdited byROBERT G. TARDIFF

EA Engineering, Science, & Technology, Inc., Silver Spring, Maryland,USA

PAULH.M.LOHMANLeiden University, The NetherlandsGERALD N. WOGAN

Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

Prepared byScientific Group on Methodologies for the Safety Evaluation of Chemicals(SGOMSEC)

Published on behalf of the Scientific Committee on the Problems of the Environment (SCOPE)of the International Council of Scientific Unions (ICSU), and the International Programme onChemical Safety (IPCS) of the World Health Organization (WHO), the United NationsEnvironment Programme (UNEP), the the International Labour Organization (ILO)

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byJOHN WILEY & SONS

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Copyright @ 1994 by theScientific Committee on Problems of the Environment (SCOPE)

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Library of Congress Cataloging-in-Publication Data

Methods to assess DNA damage and repair: interspecies comparisons /edited by Robert G. Tardiff, Paul H. M. Lohman, Gerald N. Wogan:prepared by Scientific Group on Methodologies for the SafetyEvaluation of Chemicals (SGOMSEC).

p. em. - (SCOPE; 52) (IPCS joint activity; 19) (SGOMSEC; 8)

Includes bibliographical references and index.ISBN 0-471-94256-11. DNA repair. 2. DNA damage. 3. Genetic toxicology.

4. Physiology, Comparative. I. Tardiff, Robert G. II. Lohman,Paul H. M. III. Wogan, Gerald Norman, 1930- . IV. ScientificGroup on Methodologies for the Safety Evaluation of Chemicals.V. Series: SCOPE report; 52. VI. Series: IPCS joint symposia;19. VII. Series: SGOMSEC (Series) ; 8QH467.M48 1994591.87'3282-dc20 93-37549

CIP

British Library Cataloguing in Publication Data

A catalogue record for this book is availablefrom the British Library

ISBN 0-471-94256-1

Produced from author's camera-ready copyPrinted and bound in Great Britain by Biddies Ltd, Guildford and King's Lynn

International Council of Scientific Unions (ICSU)Scientific Committee on Problems of the Environment (SCOPE)

SCOPE is one of a number of committees established by a non-governmental groupof scientific organizations, the International Council of Scientific Unions (ICSU).The membership of ICSU includes representatives from 93 National Academies ofScience, 23 International Unions, and 29 other bodies called Scientific Associates.To cover multidisciplinary activities which include the interest of several unions,ICSU has established 12 scientific committees, of which SCOPE, founded in 1969,is one. Currently, representatives of 36 member countries and 22 Unions andScientific Committees participate in the work of SCOPE, which directs particularattention to the needs of developing countries.

The mandate of SCOPE is to assemble, review, and assess the informationavailable on man-made environmental changes and the effects of these changes onman; to assess and evaluate the methodologies of measurement of environmentalparameters; to provide an intelligence service on current research; and by therecruitment of the best available scientific information and constructive thinking toestablish itself as a corpus of informed advice for the benefit of centres offundamental research and of organizations and agencies operationally engaged instudies of the environment.

SCOPE is governed by a General Assembly, which meets every three years.Between such meetings its activities are directed by the Executive Committee.

R. E. MunnEditor-in-ChiefSCOPE Publications

Executive Director: Ms V. Plocq-Fichelet

Secretariat: 51 BId de Montmorency75016 PARIS, France

Contents

Foreword Michel 1. Mercier xiii

Preface Philippe Bourdeau and Bernard D. Goldstein xv

Acknowledgements xvii

Scientific Group on Methodologies for the Safety Evaluation ofChemicals xix

Participants of the Workshop xxi

PART 1 JOINT REPORT

1 Introduction, General Conclusions, and Recommendations1.1 Introduction1.2 Carcinogenesis as a MultiStep Process1.3 Reaction Kinetics and Adduct Monitoring1.4 General Recommendations1.5 References

334568

2 DNA Damage2.1 Molecular Dosimetry

2.1.1 Absorption and Distribution2.1.2 Bioactivation and Detoxification2.1.3 Methods for Molecular Dosimetry2.1.4. Examples of Carcinogen Dosimetry in Humans

2.2 Cross-Species Sensitivity in Carcinogenic Response2.3 Approaches to Predict Cancer Activity and Potency

2.3.1 Structure-Activity Relationships2.3.1.1 Fragmentation of the Chemistry2.3.1.2 Fragmentation of Biological Parameters

2.4 Genotoxicity ProfIles in Eukaryotes

999

101112151516161718

3 DNA Repair and Mutagenesis3.1 State of Knowledge3.2 Impacts of Preferential Repair3.3 Mechanisms of Mutagenesis

19192021

Vlll CON1ENTS

4 DNA Repair, Mutagenesis, and Risk Assessment4.1 Introduction4.2 Monitoring of Human Populations

PART 2 CONTRIBUTED CHAPTERS

5 DNA Adducts and Their ConsequencesF. A. Beland and M. C. Poirier5.1 Introduction5.2 N-Nitrosamines5.3 Aflatoxins5.4 Aromatic Arnines5.5 Polycyclic Aromatic Hydrocarbons5.6 Summary5.7 References

6 Nucleotide Excision-Repair Among SpeciesJ. H. J. Hoeijmakers and A. R. Lehman6.1 Introduction6.2 Excision-Repair in Escherichia coli6.3 Excision-Repair in Saccharomyces cerevisiae

6.3.1 Yeast as a Model System6.3.2 Repair Mutants in S. cerevisiae

6.3.2.1 The RAD3 Epistasis Group6.4 Drosophila

6.4.1 Drosophila as a Model System6.4.2 Repair-Deficient Mutants6.4.3 Interaction of Excision-Repair Enzymes with Other Processes

6.4.3.1 DNA Replication on Damaged Templates6.4.3.2 Recombination

6.5 Mammalian Cells6.5.1 Cellular Biochemistry6.5.2 Repair-Deficient Mutants

6.5.2.1 Human Mutants6.5.2.2 Rodent Cell Mutants6.5.2.3 Repair of (6-4) Photoproducts6.5.2.4 Cloning of Human Repair Genes

6.6 Relationship between Repair Systems of Different Organisms6.6.1 Prokaryotes and Eukaryotes6.6.2 Yeast and Humans

6.6 References

232324

29

29303335394244

57

5757595960606363636565656565666668697074747677

CONTENTS

7 Interspecies Determinants:Bioactivation and Inactivation of CarcinogensG. 1. Mulder7.1 Introduction7.2 Exposure and Pharmacokinetic Scaling7.3 Activation of Carcinogens

7.3.1 Reactive Intermediates: Rate of Formation7.3.2 Bioactivation Mechanisms: Identification of

Ultimate Carcinogens7.4 Prediction of Metabolite Pattern

7.5 Protective Systems7.6 Extrapolation from Laboratory Animals to Humans7.7 References

8 Monitoring Cytogenetic Damage In VivoA. T. Natarajan, 1. D. Tucker, and M. S. Sasaki8.1 Introduction8.2 Chromosomal Aberrations8.3 Sister Chromatid Exchanges8.4 Micronuclei

8.4.1 Erythrocytes8.4.1.1 Bone Marrow8.4.1.2 Peripheral Blood8.4.1.3 Automation

8.4.2 Nucleated Cells8.4.2.1 Application of the CB Procedure In Vitro8.4.2.2 Application of the CB Procedure In Vivo8.4.2.3 Automation8.4.2.4 Aneuploidy

8.5 Sperm Cytogenetics8.6 Interspecies Comparisons8.7 Conclusions8.8 References

9 In Vivo Somatic Cell Gene Mutations in HumansR.1. Albertini, J. A. Nicklas, S. H. Rebison, and J. P. O'Neill9.1 Introduction

9.1.1 Rationale9.1.2 Historical Context9.1.3 Current Era

9.2 Current Assays for Human In Vivo Somatic Cell Mutations9.2.1 RBC Assays9.2.2 Hemoglobin Mutants9.2.3 G1ycophorinA (GPA) Loss Mutants

ix

83

83848585

8788919192

95

959698999999

100101101102102103104106108108109

117

117117117118118118119120

x

9.2.4 Lymphocyte Assays9.2.4.1 hpri Mutants

9.3 Other Measures of In Vivo Somatic Cell Gene Mutation9A Conclusions9.5 References

10 DNA Repair in Specific Sequences and Genomic RegionsL. F. Mullenders and C. A. Smith10.1 Introduction10.2 Repair of UV-Induced Lesions

10.2.1 Cyclobutane Pyrimidine Dimers10.2.1.1 Rodent Cells10.2.1.2 Human Cells10.2.1.3 Drosophila melanogaster10.2.1.4 Yeast10.2.1.5 E. coli

10.2.2 (6-4) Photoproducts10.2.3 Repair Heterogeneity Determined from Repair Synthesis

10.3 Alterable Repair of UV Damage in Mammalian GeneslOA Repair of Other Types of Damage

1004.1 Aflatoxin B] Adducts1004.2 Acetylaminofluorene Adducts10.4.3 Psoralen Adducts100404Adducts of Simple Alkylating Agents

10.5 Conclusions10.6 References

11 Molecular Analysis of Mutations in Endogenous GenesH. Vrieling and A. A. van Zeeland11.1 Introduction11.2 Factors Influencing Mutation Spectra

11.2.1 DNA Lesions11.2.2 DNA Structure and Sequence11.2.3 DNA Repair

11.3 Target Genes for Analysis of Mutation Spectra11.3.1 Introduction11.3.2 Chromosomal Target Genes for Mutation Studies11.3.3 Spontaneously Occurring Mutations11.304 Alkylating Agents11.3.5 Cross-linking Agents

1104 Conclusions11.5 References

CONIENTS

122122129132133

141

141142142142144145145146146147150151152152153154155156

161

161161161162163164164164165166168170170

CONTENTS

12 Molecular Analysis of Mutations in Shuttle Vectors and TransgenicAnimals 179

M. L. Wood, S. B. Verghis, and 1. M. Essigman12.1 Introduction12.2 Operational Features of Shuttle Vector and Transgenic

Animal Assays for Mutagenesis12.2.1. Transiently Replicating, Episomal Shuttle Vectors12.2.2 Stably Replicating Episomal Vectors12.2.3 Chromosomally Integrated Shuttle Vectors

12.3 Mutational Specificity of Ultraviolet Light12.4 Mutational Specificity of DNA Alkylating Agents

12.4.1 Conventional Shuttle Vector Systems12.4.2 Transgenic Animal Models12.4.3 Site-Specific Mutagenesis by Individual Alkyl-DNA Adducts

12.5 Mutational Specificity of Aromatic Amines, Amides, andNitro Compounds

12.5.1 2-Aminofluorene Derivatives12.5.2 Nitropyrene Derivatives12.5.3 N-methyl-4-Aminoazobenzene Derivatives

12.6 Mutational Specificity of Polycyclic Aromatic Hydrocarbons12.6.1 Benzo[a]pyrene12.6.2 Benzo[c]phenanthrene

12.7 Mutational Specificity of Other DNA Damaging Agents12.7.1 Iron(III)/Hydrogen Peroxide/EDTA12.7.2 Safrole12.7.3 8-Methoxypsoralen (8-MOP)

12.8 Conclusions12.9 References

13 Structure-Activity Relationships (SAR): Computerized SystemsM. D. Waters, A. M. Richards, J. R. Rabinowitz, H. F. Stack,P. H. M. Lohman, and H. S. Rosenkranz13.1 Introduction13.2 SAR as a Process13.3 Correlative and Mechanistic Approaches13.4 Correlative SAR Techniques

13.4.1 The Case System13.4.2 Chemical Classification and Representation13.4.3 Sar Modeling of Genotoxicity and Carcinogenesis

13.5 Genetic Toxicology Databases: the Gene-Tox andNTP Databases

13.5.1 Genetic Activity Profiles13.5.2 Problemswith SomeExisting Data Bases

13.6 Integration of Structural Concepts with Database Evaluation

Xl

179

180180181181182184185185186

187187188189189189190190190191191192193

201

201202204205207208209

211212213215

Xll

13.7 SAR Estimations

13.7.1 Profile Matching13.8 Weight-of-Evidence Scoring System13.9 Summary and Discussion13.10 References

14 Structure-Activity Relationships: Experimental ApproachesE. W. Vogeland 1. Ashby14.1 Introduction14.2 Early Approaches14.3 Criteria and Methods for Interspecies Comparisons

14.3.1 Genotoxic Effectiveness14.3.2 Genotoxic Efficiency14.3.3 Relative Genotoxic Efficiency

14.3.3.1 Relative Carcinogenic Potency14.3.3.2 Altered DNA Repair14.3.3.3 Biological Dosimetry

14.4 Molecular Genetic Spectra of Procarcinogens14.5 SAR Among Carcinogens/Mutagens in General14.6 Conclusions and Perspectives14.7 References

Index

CONTENTS

215217220222226

231

231231234235235235235239241245246248250

255

Foreword

The development of scientifically sound methodology for the assessment of humanhealth and environmental risks from exposure to chemicals continues to be one ofthe objectives of the International Programme on Chemical Safety (IPCS). Thisobjective also supports a major IPCS activity, namely: the preparation anddissemination of evaluations of the risks to human health and the environment fromexposure to chemicals.

Many chemicals, both man-made and natural, are capable of reacting with DNA,thus having the potential to cause cancer, mutations and adverse reproductiveoutcomes. The complexity of the field and the rapid development of knowledgerelated to DNA damage and repair makes it extremely difficult to integrate thisinformation into public health programmes. However, this can be accomplishedwhen research scientists are given an opportunity to discuss their findings withscientists having the responsibility for the assessment and management of humanhealth risks from exposure to genotoxic chemicals. By convening the workshop on"Methods to Assess DNA Damage and Repair: Interspecies Comparison," theScientific Group on Methodologies for the Safety Evaluation of Chemicals(SGOMSEC) has provided such an opportunity.

The in-depth scientific review made by world leaders in the field will providemuch needed guidance to those asked to use the results from studies inexperimental animals to assess human health risks. Also, the results of thisSGOMSEC activity will assist IPCS in developing further its activities related tothe role of biomarkers in the overall process of health risk assessment.

Michel J. MercierManager, InternationalProgramme on Chemical Safety

Preface

The Scientific Group on Methodologies for the Safety Evaluation of Chemicals(SGOMSEC) was established in 1979 at the initiative of Professor Norton Nelsonfrom New York University. It is a non-governmental organization sponsored byIPCS (the International Program on Chemical Safety, established within WHO withthe cooperation of UNEP and ILO), and the Scientific Committee on Problems ofthe Environment (SCOPE), itself a body of the International Council of ScientificUnions (ICSU).

The broad objective of SGOMSEC is to contribute to the reduction andprevention of risks caused to humans and non-human targets (ecosystems) by theintroduction in the environment in increasing quantities of a large number of naturaland man-made chemicals. The specific contribution of SGOMSEC is to assess themethodologies in use for the evaluation of these risks with a view to determinetheir values, to identify gaps and emerging needs, and to make recommendationsfor future research. Previous SGOMSEC projects have dealt either with generalproblems of chemical exposure and effects or with specific issues such as the non-intentional effects of pesticides or the consequences of large chemical accidents.

Volume 8 of SGOMSEC is concerned with a subject basic to chemical riskassessment: DNA damage and repair, taking account of interspecies differences.Many natural and man-made chemicals, as well as some physical agents, may reactwith DNA and thus produce harmful effect including cancer. However, mammalianspecies have various defence systems which may overcome the deleterious effects,of these genotoxic agents: metabolic and pharmacokinetic processes, DNA repairmechanisms, immunological processes, etc. This report evaluates currentknowledge of the mechanisms of activation/detoxication of DNA damaging agents,organotropic and cell-structure effects, induction and repair of DNA damage, andthe molecular and phenotypical analysis of mutation induction This evaluation ispresented in seven contributed papers and a joint report which was prepared at aworkshop held at the National Institute of Environmental Health Sciences (NIEHS),Research Triangle Park, North Carolina, USA in March 1990.

The joint report includes general recommendations aimed at improving researchin this field and developing methodologies for genotoxicity testing, taking accountof the potentials and limitations of available animal models.

The project was co-chaired by Professors H. M. Lohman of Leiden Universityand Gerald N. Wogan of MIT. They and the authors of the contributed papers, allexperts in their respective fields, are thanked for their participation. We are alsomost grateful to Robert G. Tardiff (EA Engineering, Science and Technology, Inc.,Silver Spring, Maryland) for his part in editing the report.

Special thanks are due to Dr. David Rall, Director of NIEHS, for hosting the

XVI PREFACE

workshop at the US National Institute of Environmental Health Sciences (NIEHS).SGOMSEC gratefully acknowledges the support and financial assistance ofIPCS,

SCOPE, NIEHS, the US Environmental Protection Agency, and the Commissionof the European Community, without which this project would not have beenpossible.

Philippe BourdeauBernard D. Goldstein

Co-Chairmen, Scientific Groupon Methodologies for theSafety Evaluation of Chemicals

Acknowledgements

The editors are pleased to acknowledge the assistance received from:

J. GalvinI. GobelC. HartwickM. JacksonS. JohnsonE. LinthicumT. PiccinP. RoneyC. StapletonB. TardiffL. Wakefield

Scientific Group onMethodologies for the SafetyEvaluation of Chemicals

*George C. BeckingInterregional Research Unit, International Programme on Chemical Safety, WorldHealth Organization, MD-A206, P.O. Box /2233, Research Triangle Park, NC27709 USA (Secretary)

N. P. Bockov

Director, Institute of Medical Genetics, Kasirskoesosse 6A Moscow 115478, USSR

*Philippe BourdeauDirectorate General for Science Research and Development, Commission of theEuropean Communities, Rue de la Loi 200, B-1049 Brussels, Belgium (Chairman)

*Bernard Goldstein

Professor and Chairman, Department of Environmental and Community Medicine,UMDNJ-RobertWoodJohnsonMedicalSchool,675Hoes Lane,Piscataway,NJ08854, USA (Vice Chairman)

Gareth GreenProfessor and Chairman,Department of Environmental Health Sciences, The JohnsHopkins University, School of Hygiene and Public Health, 615 North Wolfe Street,Baltimore, MD 21205 USA

*Miki GotoProfessor, Department of Chemistry and Director, Institute of Ecotoxicology,Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171, Japan

J. R. HickmanBureauof ChemicalHazards,EnvironmentalHealthCenter,DepartmentHealthand Welfare,Tunney'sPasture,Ottawa,OntarioKLAOL2,Canada

Vladimir LandaCzechoslovak Academy of Sciences, Institute of Entomology, Branisovdska 31,370

05 Ceske Budejovice, Czechoslovakia Republic

xx SCIENTIFIC GROUP ON METHODOLOGIES

* Aly Massoud

Professor and Chairman, Department of Community, Environmental andOccupational Medicine, Ein Shams University, Abbassia Cairo, Egypt

*Michel MercierManager, International Programme on Chemical Safety, World HealthOrganization, 12/I Geneva 27, Switzerland

Thomas OdhiamboDirector, International Centrefor Insect Physiology and Ecology, P.O. Box 30772,Nairobi, Kenya

*Blanca Raquel OrdonezEnvironmental Advisor of the Minister of Health, School of Public Health, LopezCotilla 739, Col. del Valle, 03/00 D.F. Mexico

Dennis V. ParkeHead, Department of Biochemistry, University of Surrey UK, Guildford, SurreyGU2 5XH, UK

David B. PeakaUMonitoring and Research Centre, Old Coach House, Campden Hill, London, UK

Jerzy PiotrowskiLodz Medical Academy, Institute of Environmental Research, Narutowicza I20A,90-145 Lodz, Poland

David P. Rail5302 Reno Road, NW, Washington, DC 20015, USA

*Robert G. TardiffVice President for Health Sciences, EA Engineering, Science, & Technology, Inc.,8401 Colesville Road, Silver Spring, MD 20910, USA (Editor)

Rene TruhautLaboratoire de Toxicologie et d'Hygiene Industrielle, Faculte des Sciences,Pharmaceutiques et Biologiques de Paris, Universite Rene Descartes, 4 avenue deI'Observatoire, 75006 Paris, France

*Member of the Executive Committee