Susceptibility towards formaldehyde-induced genotoxicity

Gnter Speit
Universitt Ulm Institut fr Humangenetik D-89069 Ulm (Germany) guenter.speit@uni-ulm.de

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Susceptibility towards formaldehyde-induced genotoxicity

Outline of the presentation:

I. II. III. IV. V. Introduction Nasal sensitivity and susceptibility towards genotoxicity Potential susceptibility of different study groups Polymorphisms in the FDH gene and susceptibility Summary

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I. Introduction

Susceptible subgroups are groups of individuals with predisposition to greater response to an exposure: Genetics (e.g., genetic polymorphisms) Gender differences Life stages (e.g. children) Health status (e.g., asthmatics) Lifestyle factors Nutritional state Exposure to other chemicals

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Background

SCOEL (2008): Exposure limit should especially consider possible inter-individual differences in susceptibility to irritation by FA. The critical health effect of exposure to FA is the potential to induce cancer in the upper respiratory tract. There is no reason to assume that subgroups with increased susceptibility to irritation may also have a higher risk for the induction of mutations and cancer but data are not available. We investigated the in vitro genotoxic susceptibility of blood cells in relationship to nasal sensitivity.

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Cellular defence against FA-induced mutagenicity

methanol

endogenous sources

exogenous sources

DPX

Repair

formaldehyde
glutathione adduct formation

Proliferation

NAD+ ALDH1A1 ALDH2 NADH +H+

S-hydroxymethylglutathione
NAD+

H2O2

Mutation

FDH
NADH +H+

one carbon pool

catalase

H2O

S-formylglutathione
S-formylglutathione hydrolase

glutathione

formate

Increased susceptibility may be due to: - impaired metabolic inactivation - impaired DNA repair

CO2 + H2O

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II. Nasal sensitivity and susceptibility towards genotoxicity

Study design: 41 volunteers (male non-smokers) Definition of hyper- and hyposensitive subgroups with regard to unspecific nasal irritation (CO2-sensitivity). Determination of cellular protection by FDH (gene expression) Determination of cellular susceptibility to DNA damage and cellular repair capacity by tests with peripheral blood as a surrogate tissue for studying mutagen sensitivity.

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II. Nasal sensitivity and susceptibility towards genotoxicity

Blood samples were exposed to FA in vitro: Comet assay was performed to measure - FA-induced DNA damage (DPX) - removal (repair) of FA-induced DPX

SCE-test was performed to measure - persistence of FA-induced DNA damage - FA-induced genotoxicity (SCE)

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II. Nasal sensitivity and susceptibility towards genotoxicity

Results: Large differences in CO2-sensitivity (nasal irritation) No significant differences between the subgroups - with regard to the induction of genotoxic effects (DPX, SCE) - with regard to the protection against FA (FDH; DPX-repair)
Zeller et al. Mutat. Res. 723, 11-17 (2011)
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II. Nasal sensitivity and susceptibility towards genotoxicity

Correlation analysis for all parameters tested: - for all subjects, - for the subgroups of hypersensitive and hyposensitive subjects, - for the subgroups of extremly hypersensitive and hyposensitive subjects. Results: No correlation between CO2-sensitivity and the expression of FDH. No correlation between CO2-sensitivity and indicators of cellular sensitivity. No subgroup with particular mutagen sensitivity towards FA was identified.
Zeller et al. Mutat. Res. 723, 11-17 (2011)
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III. Potential susceptibility of different study groups

Ex-vivo susceptibility towards FA-induced genotoxicity was tested: - in 30 male smokers - in 30 female non-smokers - in 30 school children The comet assay and the SCE-test were performed in accordance with the previous study. The expression of the FDH-gene was measured by quantitative real-time RT-PCR. Polymorphisms in GSTM1 and GSTT1 were considered.
Zeller et al. Arch. Toxicol., in press (2012)
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III. Potential susceptibility of different study groups

No biologically relevant differences between the three groups.

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III. Potential susceptibility of different study groups

DNA damage

SCE-test

DNA repair

FDH

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III. Potential susceptibility of different study groups

Induced genotoxic effects (comet assay) are not associated with polymorphisms in GSTM1 and GSTT1.

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III. Potential susceptibility of different study groups

Induced SCE frequencies are slightly higher in subjects with GSTM1 null genotype. Induced SCE frequencies are not associated with polymorphisms in GSTT1 and GSTM1/GSTT1.
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III. Potential susceptibility of different study groups

Summary No biologically relevant differences were measured between the three study groups with regard to the tested indicators of cellular sensitivity towards FA-induced genotoxic effects and the expression of FDH. There was no clear association between the induced genotoxic effects and polymorphisms in GSTM1 and GSTT1. None of the groups showed particular mutagen sensitivity towards FA-induced genotoxicity. The results suggest that a low scaling factor to address possible human inter-individual differences in FA-induced mutagenicity could be reasonable.
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IV. Polymorphisms in the FDH gene and susceptibility

We identified three SNPs in the FDH-gene (ADH5) with a potential impact on gene expression (NCBI database). No variant allel was detected for 2 SNPs in our study population. 89 Subjects with a variant allele were identified for rs13832 (exon 9).

Just et al. Toxicol. Lett. 207, 121-127 (2011)

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IV. Polymorphisms in the FDH gene and susceptibility

The variant allele did not cause a difference in FDH expression (mRNA level).

The variant allele did not cause a difference in the protection of cells against FA-induced genotoxicity (comet assay): - No difference in background DNA effects - No difference in FA-induced DPX No indication for inter-individual differences in the metabolic inactivation of FA.
Just et al. Toxicol. Lett. 207, 121-127 (2011)
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Susceptibility towards FA-induced genotoxicity

Summary Our study did not reveal a polymorphism of FDH which might affect gene expression and lead to inter-individual differences in the protection of FA-induced (geno-)toxicity. Altogether, our studies do not point to relevant differences in susceptibility towards FA-induced genotoxicity which need to be considered in risk assessment.

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THANK YOU
Acknowledgements: Jasmin Zeller Alexandra Ulrich Clarissa Riegert Walter Just Simone Neu Christopher Teller Regina Linsenmeyer Josef Hgel Gerhard Triebig Jrg U. Mller Thomas Bruckner

Financial support: EPF & CEFIC / FormaCare

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