TY - JOUR
T1 - Metabolic Activation of Benzo[a]pyrene by Human Tissue Organoid Cultures
AU - Caipa Garcia, Angela L.
AU - Kucab, Jill E.
AU - Al-Serori, Halh
AU - Beck, Rebekah S.S.
AU - Fischer, Franziska
AU - Hufnagel, Matthias
AU - Hartwig, Andrea
AU - Floeder, Andrew
AU - Balbo, Silvia
AU - Francies, Hayley
AU - Garnett, Mathew
AU - Huch, Meritxell
AU - Drost, Jarno
AU - Zilbauer, Matthias
AU - Arlt, Volker M.
AU - Phillips, David H.
N1 - Funding Information:
This work was supported by the UK Medical Research Council (MR/N013700/1 to A.L.C.G.) and King’s College London, which is a member of the MRC Doctoral Training Partnership in Biomedical Sciences, and by Cancer Research UK Grand Challenge Award “Mutographs of Cancer” (C98/A24032). D.H.P. is a member of the Health Protection Research Unit in Chemical and Radiation Threats and Hazards, a partnership with UK Health Security Agency and Imperial College London, which is funded by the National Institute for Health Research (NIHR).
Publisher Copyright:
© 2022 by the authors.
PY - 2023/1
Y1 - 2023/1
N2 - Organoids are 3D cultures that to some extent reproduce the structure, composition and function of the mammalian tissues from which they derive, thereby creating in vitro systems with more in vivo-like characteristics than 2D monocultures. Here, the ability of human organoids derived from normal gastric, pancreas, liver, colon and kidney tissues to metabolise the environmental carcinogen benzo[a]pyrene (BaP) was investigated. While organoids from the different tissues showed varied cytotoxic responses to BaP, with gastric and colon organoids being the most susceptible, the xenobiotic-metabolising enzyme (XME) genes, CYP1A1 and NQO1, were highly upregulated in all organoid types, with kidney organoids having the highest levels. Furthermore, the presence of two key metabolites, BaP-t-7,8-dihydrodiol and BaP-tetrol-l-1, was detected in all organoid types, confirming their ability to metabolise BaP. BaP bioactivation was confirmed both by the activation of the DNA damage response pathway (induction of p-p53, pCHK2, p21 and γ-H2AX) and by DNA adduct formation. Overall, pancreatic and undifferentiated liver organoids formed the highest levels of DNA adducts. Colon organoids had the lowest responses in DNA adduct and metabolite formation, as well as XME expression. Additionally, high-throughput RT-qPCR explored differences in gene expression between organoid types after BaP treatment. The results demonstrate the potential usefulness of organoids for studying environmental carcinogenesis and genetic toxicology.
AB - Organoids are 3D cultures that to some extent reproduce the structure, composition and function of the mammalian tissues from which they derive, thereby creating in vitro systems with more in vivo-like characteristics than 2D monocultures. Here, the ability of human organoids derived from normal gastric, pancreas, liver, colon and kidney tissues to metabolise the environmental carcinogen benzo[a]pyrene (BaP) was investigated. While organoids from the different tissues showed varied cytotoxic responses to BaP, with gastric and colon organoids being the most susceptible, the xenobiotic-metabolising enzyme (XME) genes, CYP1A1 and NQO1, were highly upregulated in all organoid types, with kidney organoids having the highest levels. Furthermore, the presence of two key metabolites, BaP-t-7,8-dihydrodiol and BaP-tetrol-l-1, was detected in all organoid types, confirming their ability to metabolise BaP. BaP bioactivation was confirmed both by the activation of the DNA damage response pathway (induction of p-p53, pCHK2, p21 and γ-H2AX) and by DNA adduct formation. Overall, pancreatic and undifferentiated liver organoids formed the highest levels of DNA adducts. Colon organoids had the lowest responses in DNA adduct and metabolite formation, as well as XME expression. Additionally, high-throughput RT-qPCR explored differences in gene expression between organoid types after BaP treatment. The results demonstrate the potential usefulness of organoids for studying environmental carcinogenesis and genetic toxicology.
KW - 3D culture
KW - benzo[a]pyrene
KW - carcinogen
KW - CYP1A1
KW - DNA adducts
KW - DNA damage response
KW - human tissue organoid
KW - NQO1
KW - RT-qPCR
UR - http://www.scopus.com/inward/record.url?scp=85145981245&partnerID=8YFLogxK
U2 - 10.3390/ijms24010606
DO - 10.3390/ijms24010606
M3 - Article
C2 - 36614051
AN - SCOPUS:85145981245
SN - 1661-6596
VL - 24
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 1
M1 - 606
ER -