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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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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 |
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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 |
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