Application of the comet assay in human biomonitoring: An hCOMET perspective.

Amaya Azqueta, Carina Ladeira, Lisa Giovannelli, Elisa Boutet-Robinet, Stefano Bonassi, Monica Neri, Goran Gajski, Susan Duthie, Cristian Del Bo', Patrizia Riso, Gudrun Koppen, Nursen Basaran, Andrew Collins, Peter Møller

Journal: Mutation research. Reviews in mutation research 2020;783():108288

PMID: 32192646

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Abstract

The comet assay is a well-accepted biomonitoring tool to examine the effect of dietary, lifestyle, environmental and occupational exposure on levels of DNA damage in human cells. With such a wide range of determinants for DNA damage levels, it becomes challenging to deal with confounding and certain factors are inter-related (e.g. poor nutritional intake may correlate with smoking status). This review describes the effect of intrinsic (i.e. sex, age, tobacco smoking, occupational exposure and obesity) and extrinsic (season, environmental exposures, diet, physical activity and alcohol consumption) factors on the level of DNA damage measured by the standard or enzyme-modified comet assay. Although each factor influences at least one comet assay endpoint, the collective evidence does not indicate single factors have a large impact. Thus, controlling for confounding may be necessary in a biomonitoring study, but none of the factors is strong enough to be regarded a priori as a confounder. Controlling for confounding in the comet assay requires a case-by-case approach. Inter-laboratory variation in levels of DNA damage and to some extent also reproducibility in biomonitoring studies are issues that have haunted the users of the comet assay for years. Procedures to collect specimens, and their storage, are not standardized. Likewise, statistical issues related to both sample-size calculation (before sampling of specimens) and statistical analysis of the results vary between studies. This review gives guidance to statistical analysis of the typically complex exposure, co-variate, and effect relationships in human biomonitoring studies.

Address: Department of Pharmacology and Toxicology, University of Navarra, C/Irunlarrea 1, 31009 Pamplona, Spain; IdiSNA, Navarra Institute for Health Research, Spain.; H&TRC - Health & Technology Research Center, ESTeSL - Escola Superior de Tecnologia da Saúde, Instituto Politécnico de Lisboa, Av. D. João II, lote 4.69.01, Parque das Nações, 1990-096 Lisboa, Portugal; Centro de Investigação e Estudos em Saúde Pública, Escola Nacional de Saúde Pública, ENSP, Universidade Nova de Lisboa, Portugal.; Dept. NEUROFARBA, Section Pharmacology and Toxicology, University of Florence, Florence, Italy.; Toxalim (Research Centre in Food Toxicology), Université de Toulouse, INRA, ENVT, INP-Purpan, UPS, Toulouse, France.; Unit of Clinical and Molecular Epidemiology IRCCS San Raffaele Pisana, Rome, Italy; Department of Human Sciences and Quality of Life Promotion, San Raffaele University, Rome, Italy.; Unit of Clinical and Molecular Epidemiology IRCCS San Raffaele Pisana, Rome, Italy.; Mutagenesis Unit, Institute for Medical Research and Occupational Health, Ksaverska Cesta 2, 10000 Zagreb, Croatia.; School of Pharmacy and Life Sciences, The Robert Gordon University, Riverside East, Garthdee Road, Aberdeen AB10 7GJ, United Kingdom.; Università degli Studi di Milano, Department of Food, Environmental and Nutritional Sciences (DeFENS), Division of Human Nutrition, Via G. Celoria 2, 20133 Milano, Italy.; VITO Health, Flemish Institute for Technological Research (VITO), Mol, Belgium.; Hacettepe University, Faculty of Pharmacy, Department of Toxicology, 06100 Ankara, Turkey.; Department of Nutrition, Institute for Basic Medical Sciences, University of Oslo, Sognsvannsveien 9, 0372 Oslo, Norway.; Department of Public Health, Section of Environmental Health, University of Copenhagen, DK-1014 Copenhagen K, Denmark.

DOI: 10.1016/j.mrrev.2019.108288

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MeSH Terms: Adult, Age Factors, Biological Monitoring, Comet Assay, DNA Damage, DNA-Formamidopyrimidine Glycosylase, Environmental Exposure, Escherichia coli Proteins, Female, Humans, Male, Middle Aged, Obesity, Oxidative Stress, Risk Factors, Seasons, Sex Factors, Tobacco Smoking