The risk of exposure to nanoparticles (NPs) has rapidly increased during the last decade due to the vast use of nanomaterials (NMs) in many areas of human life. Despite this fact, human biomonitoring studies focused on the effect of NP exposure on DNA alterations are still rare. Furthermore, there are virtually no epigenetic data available. In this study, we investigated global and gene-specific DNA methylation profiles in a group of 20 long-term (mean 14.5 years) exposed, nanocomposite, research workers and in 20 controls. Both groups were sampled twice/day (pre-shift and post-shift) in September 2018. We applied Infinium Methylation Assay, using the Infinium MethylationEPIC BeadChips with more than 850,000 CpG loci, for identification of the DNA methylation pattern in the studied groups. Aerosol exposure monitoring, including two nanosized fractions, was also performed as proof of acute NP exposure. The obtained array data showed significant differences in methylation between the exposed and control groups related to long-term exposure, specifically 341 CpG loci were hypomethylated and 364 hypermethylated. The most significant CpG differences were mainly detected in genes involved in lipid metabolism, the immune system, lung functions, signaling pathways, cancer development and xenobiotic detoxification. In contrast, short-term acute NP exposure was not accompanied by DNA methylation changes. In summary, long-term (years) exposure to NP is associated with DNA epigenetic alterations.

Center for Experimental Medicine Institute for Clinical and Experimental Medicine Videnska 1958 9 140 21 Prague 4 Czech Republic

Department of Computer Science Czech Technical University Prague Karlovo namesti 13 121 35 Prague 2 Czech Republic

Department of Genetic Toxicology and Epigenetics Institute of Experimental Medicine CAS Videnska 1083 142 20 Prague 4 Czech Republic

Department of Machining and Assembly Department of Engineering Technology Department of Material Science Faculty of Mechanical Engineering Technical University in Liberec Studentska 1402 2 Liberec Czech Republic

Department of Nanotoxicology and Molecular Epidemiology Institute of Experimental Medicine CAS Videnska 1083 142 20 Prague 4 Czech Republic

Department of Occupational Medicine 1st Faculty of Medicine Charles University Prague and General University Hospital Prague Na Bojisti 1 120 00 Prague 2 Czech Republic

Laboratory of Aerosol Chemistry and Physics Institute of Chemical Process Fundamentals CAS Rozvojova 1 165 02 Prague 6 Czech Republic

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