Human tissue three-dimensional (3D) organoid cultures
have the
potential to reproduce in vitro the physiological
properties and cellular architecture of the organs from which they
are derived. The ability of organoid cultures derived from human stomach,
liver, kidney, and colon to metabolically activate three dietary carcinogens,
aflatoxin B1 (AFB1), aristolochic acid I (AAI),
and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine
(PhIP), was investigated. In each case, the response of a target tissue
(liver for AFB1; kidney for AAI; colon for PhIP) was compared
with that of a nontarget tissue (gastric). After treatment cell viabilities
were measured, DNA damage response (DDR) was determined by Western
blotting for p-p53, p21, p-CHK2, and γ-H2AX, and DNA adduct
formation was quantified by mass spectrometry. Induction of the key
xenobiotic-metabolizing enzymes (XMEs) CYP1A1, CYP1A2, CYP3A4, and
NQO1 was assessed by qRT-PCR. We found that organoids from different
tissues can activate AAI, AFB1, and PhIP. In some cases,
this metabolic potential varied between tissues and between different
cultures of the same tissue. Similarly, variations in the levels of
expression of XMEs were observed. At comparable levels of cytotoxicity,
organoids derived from tissues that are considered targets for these
carcinogens had higher levels of adduct formation than a nontarget
tissue.