Cargando…

Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids

Short-/middle-term and simple prediction studies for carcinogenesis are needed for the safety assessment of chemical substances. To establish a novel genotoxicity assay with an in vivo mimicking system, we prepared murine colonic/pulmonary organoids from gpt delta mice according to the general proce...

Descripción completa

Detalles Bibliográficos
Autores principales: Komiya, Masami, Ishigamori, Rikako, Naruse, Mie, Ochiai, Masako, Miyoshi, Noriyuki, Imai, Toshio, Totsuka, Yukari
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8638810/
https://www.ncbi.nlm.nih.gov/pubmed/34868254
http://dx.doi.org/10.3389/fgene.2021.768781
_version_ 1784609015003611136
author Komiya, Masami
Ishigamori, Rikako
Naruse, Mie
Ochiai, Masako
Miyoshi, Noriyuki
Imai, Toshio
Totsuka, Yukari
author_facet Komiya, Masami
Ishigamori, Rikako
Naruse, Mie
Ochiai, Masako
Miyoshi, Noriyuki
Imai, Toshio
Totsuka, Yukari
author_sort Komiya, Masami
collection PubMed
description Short-/middle-term and simple prediction studies for carcinogenesis are needed for the safety assessment of chemical substances. To establish a novel genotoxicity assay with an in vivo mimicking system, we prepared murine colonic/pulmonary organoids from gpt delta mice according to the general procedure using collagenase/dispase and cultured them in a 3D environment. When the organoids were exposed to foodborne carcinogens—2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) and acrylamide (AA)—in the presence of metabolic activation systems, mutation frequencies (MFs) occurring in the gpt gene dose-dependently increased. Moreover, the mutation spectrum analysis indicated predominant G:C to T:A transversion with PhIP, and A:T to C:G and A:T to T:A transversion with AA. These data correspond to those of a previous study describing in vivo mutagenicity in gpt delta mice. However, organoids derived from the liver, a non-target tissue of PhIP-carcinogenesis, also demonstrated genotoxicity with a potency comparable to colonic organoids. Organoids and PhIP were directly incubated in the presence of metabolic activation systems; therefore, there was a lack of organ specificity, as observed in vivo. Additionally, PhIP-DNA adduct levels were comparable in hepatic and colonic organoids after PhIP exposure. Taken together, the organoids prepared in the present study may be helpful to predict chemical carcinogenesis.
format Online
Article
Text
id pubmed-8638810
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-86388102021-12-03 Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids Komiya, Masami Ishigamori, Rikako Naruse, Mie Ochiai, Masako Miyoshi, Noriyuki Imai, Toshio Totsuka, Yukari Front Genet Genetics Short-/middle-term and simple prediction studies for carcinogenesis are needed for the safety assessment of chemical substances. To establish a novel genotoxicity assay with an in vivo mimicking system, we prepared murine colonic/pulmonary organoids from gpt delta mice according to the general procedure using collagenase/dispase and cultured them in a 3D environment. When the organoids were exposed to foodborne carcinogens—2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) and acrylamide (AA)—in the presence of metabolic activation systems, mutation frequencies (MFs) occurring in the gpt gene dose-dependently increased. Moreover, the mutation spectrum analysis indicated predominant G:C to T:A transversion with PhIP, and A:T to C:G and A:T to T:A transversion with AA. These data correspond to those of a previous study describing in vivo mutagenicity in gpt delta mice. However, organoids derived from the liver, a non-target tissue of PhIP-carcinogenesis, also demonstrated genotoxicity with a potency comparable to colonic organoids. Organoids and PhIP were directly incubated in the presence of metabolic activation systems; therefore, there was a lack of organ specificity, as observed in vivo. Additionally, PhIP-DNA adduct levels were comparable in hepatic and colonic organoids after PhIP exposure. Taken together, the organoids prepared in the present study may be helpful to predict chemical carcinogenesis. Frontiers Media S.A. 2021-11-18 /pmc/articles/PMC8638810/ /pubmed/34868254 http://dx.doi.org/10.3389/fgene.2021.768781 Text en Copyright © 2021 Komiya, Ishigamori, Naruse, Ochiai, Miyoshi, Imai and Totsuka. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Genetics
Komiya, Masami
Ishigamori, Rikako
Naruse, Mie
Ochiai, Masako
Miyoshi, Noriyuki
Imai, Toshio
Totsuka, Yukari
Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
title Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
title_full Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
title_fullStr Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
title_full_unstemmed Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
title_short Establishment of Novel Genotoxicity Assay System Using Murine Normal Epithelial Tissue-Derived Organoids
title_sort establishment of novel genotoxicity assay system using murine normal epithelial tissue-derived organoids
topic Genetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8638810/
https://www.ncbi.nlm.nih.gov/pubmed/34868254
http://dx.doi.org/10.3389/fgene.2021.768781
work_keys_str_mv AT komiyamasami establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids
AT ishigamoririkako establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids
AT narusemie establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids
AT ochiaimasako establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids
AT miyoshinoriyuki establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids
AT imaitoshio establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids
AT totsukayukari establishmentofnovelgenotoxicityassaysystemusingmurinenormalepithelialtissuederivedorganoids