TY - JOUR
T1 - Mutagenicity of acrylamide and glycidamide in human TP53 knock-in (Hupki) mouse embryo fibroblasts
AU - Holzl-Armstrong, Lisa
AU - Kucab, Jill
AU - Moody, Sarah
AU - Zwart, Edwin
AU - Loutkovotova, Lucie
AU - Duffy, Veronica
AU - Luijten, Mirjiam
AU - Gamboa da Costa, Goncalo
AU - Stratton, Michael
AU - Phillips, David
AU - Arlt, Volker
PY - 2020/8/12
Y1 - 2020/8/12
N2 - Acrylamide is a suspected human carcinogen formed during high-temperature cooking of starch-rich foods. It is metabolised
by cytochrome P450 2E1 to its reactive metabolite glycidamide, which forms pre-mutagenic DNA adducts. Using the human
TP53 knock-in (Hupki) mouse embryo ibroblasts (HUFs) immortalisation assay (HIMA), acrylamide- and glycidamideinduced
mutagenesis was studied in the tumour suppressor gene TP53. Selected immortalised HUF clones were also subjected
to next-generation sequencing to determine mutations across the whole genome. The TP53-mutant frequency after
glycidamide exposure (1.1 mM for 24 h, n = 198) was 9% compared with 0% in cultures treated with acrylamide [1.5 (n = 24)
or 3 mM (n = 6) for 48 h] and untreated vehicle (water) controls (n = 36). Most glycidamide-induced mutations occurred at
adenines with A > T/T > A and A > G/T > C mutations being the most common types. Mutations induced by glycidamide
occurred at speciic TP53 codons that have also been found to be mutated in human tumours (i.e., breast, ovary, colorectal,
and lung) previously associated with acrylamide exposure. The spectrum of TP53 mutations was further relected by the
mutations detected by whole-genome sequencing (WGS) and a distinct WGS mutational signature was found in HUF clones
treated with glycidamide that was again characterised by A > G/T > C and A > T/T > A mutations. The WGS mutational
signature showed similarities with COSMIC mutational signatures 3 and 25 previously found in human tumours (e.g., breast
and ovary), while the adenine component was similar to COSMIC SBS4 found mostly in smokers’ lung cancer. In contrast, in
acrylamide-treated HUF clones, only culture-related background WGS mutational signatures were observed. In summary, the
results of the present study suggest that glycidamide may be involved in the development of breast, ovarian, and lung cancer.
AB - Acrylamide is a suspected human carcinogen formed during high-temperature cooking of starch-rich foods. It is metabolised
by cytochrome P450 2E1 to its reactive metabolite glycidamide, which forms pre-mutagenic DNA adducts. Using the human
TP53 knock-in (Hupki) mouse embryo ibroblasts (HUFs) immortalisation assay (HIMA), acrylamide- and glycidamideinduced
mutagenesis was studied in the tumour suppressor gene TP53. Selected immortalised HUF clones were also subjected
to next-generation sequencing to determine mutations across the whole genome. The TP53-mutant frequency after
glycidamide exposure (1.1 mM for 24 h, n = 198) was 9% compared with 0% in cultures treated with acrylamide [1.5 (n = 24)
or 3 mM (n = 6) for 48 h] and untreated vehicle (water) controls (n = 36). Most glycidamide-induced mutations occurred at
adenines with A > T/T > A and A > G/T > C mutations being the most common types. Mutations induced by glycidamide
occurred at speciic TP53 codons that have also been found to be mutated in human tumours (i.e., breast, ovary, colorectal,
and lung) previously associated with acrylamide exposure. The spectrum of TP53 mutations was further relected by the
mutations detected by whole-genome sequencing (WGS) and a distinct WGS mutational signature was found in HUF clones
treated with glycidamide that was again characterised by A > G/T > C and A > T/T > A mutations. The WGS mutational
signature showed similarities with COSMIC mutational signatures 3 and 25 previously found in human tumours (e.g., breast
and ovary), while the adenine component was similar to COSMIC SBS4 found mostly in smokers’ lung cancer. In contrast, in
acrylamide-treated HUF clones, only culture-related background WGS mutational signatures were observed. In summary, the
results of the present study suggest that glycidamide may be involved in the development of breast, ovarian, and lung cancer.
KW - TP53 · Mutation · Whole-genome sequencing · Dietary carcinogen · Acrylamide · DNA adducts
M3 - Article
SN - 0340-5761
JO - Archives of Toxicology
JF - Archives of Toxicology
M1 - 2878
ER -
