TY - JOUR
T1 - Nanomaterial genotoxicity evaluation using the high-throughput p53-binding protein 1 (53BP1) assay
AU - Fontaine, Maelle
AU - Bartolami, Eline
AU - Prono, Marion
AU - Béal, David
AU - Blosi, Magda
AU - Costa, Anna L.
AU - Ravagli, Costanza
AU - Baldi, Giovanni
AU - Sprio, Simone
AU - Tampieri, Anna
AU - Fenoglio, Ivana
AU - Tran, Lang
AU - Fadeel, Bengt
AU - Carriere, Marie
N1 - Funding Information:
Funding: EB, MB, ALC, CR, GB, SS, AT, IF, LT, BF, MC received funding by the European Union’s Horizon 2020 research and innovation programme under grant agreement no. 760928 (BIORIMA, https://www.biorima.eu). DB, MC received funding from Labex Serenade (ANR-11-LABX-0064, https://labex-serenade.fr) funded by the French Government’s Investissements d’Avenir program of Agence Nationale de la Recherche, through the A*MIDEX project (ANR-11-IDEX-0001-02). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
Copyright: © 2023 Fontaine et al.
PY - 2023/9
Y1 - 2023/9
N2 - Toxicity evaluation of engineered nanomaterials is challenging due to the ever increasing number of materials and because nanomaterials (NMs) frequently interfere with commonly used assays. Hence, there is a need for robust, high-throughput assays with which to assess their hazard potential. The present study aimed at evaluating the applicability of a genotoxicity assay based on the immunostaining and foci counting of the DNA repair protein 53BP1 (p53-binding protein 1), in a high-throughput format, for NM genotoxicity assessment. For benchmarking purposes, we first applied the assay to a set of eight known genotoxic agents, as well as X-ray irradiation (1 Gy). Then, a panel of NMs and nanobiomaterials (NBMs) was evaluated with respect to their impact on cell viability and genotoxicity, and to their potential to induce reactive oxygen species (ROS) production. The genotoxicity recorded using the 53BP1 assay was confirmed using the micronucleus assay, also scored via automated (high-throughput) microscopy. The 53BP1 assay successfully identified genotoxic compounds on the HCT116 human intestinal cell line. None of the tested NMs showed any genotoxicity using the 53BP1 assay, except the positive control consisting in (CoO)(NiO) NMs, while only TiO2 NMs showed positive outcome in the micronucleus assay. Only Fe3O4 NMs caused significant elevation of ROS, not correlated to DNA damage. Therefore, owing to its adequate predictivity of the genotoxicity of most of the tested benchmark substance and its ease of implementation in a high throughput format, the 53BP1 assay could be proposed as a complementary high-throughput screening genotoxicity assay, in the context of the development of New Approach Methodologies.
AB - Toxicity evaluation of engineered nanomaterials is challenging due to the ever increasing number of materials and because nanomaterials (NMs) frequently interfere with commonly used assays. Hence, there is a need for robust, high-throughput assays with which to assess their hazard potential. The present study aimed at evaluating the applicability of a genotoxicity assay based on the immunostaining and foci counting of the DNA repair protein 53BP1 (p53-binding protein 1), in a high-throughput format, for NM genotoxicity assessment. For benchmarking purposes, we first applied the assay to a set of eight known genotoxic agents, as well as X-ray irradiation (1 Gy). Then, a panel of NMs and nanobiomaterials (NBMs) was evaluated with respect to their impact on cell viability and genotoxicity, and to their potential to induce reactive oxygen species (ROS) production. The genotoxicity recorded using the 53BP1 assay was confirmed using the micronucleus assay, also scored via automated (high-throughput) microscopy. The 53BP1 assay successfully identified genotoxic compounds on the HCT116 human intestinal cell line. None of the tested NMs showed any genotoxicity using the 53BP1 assay, except the positive control consisting in (CoO)(NiO) NMs, while only TiO2 NMs showed positive outcome in the micronucleus assay. Only Fe3O4 NMs caused significant elevation of ROS, not correlated to DNA damage. Therefore, owing to its adequate predictivity of the genotoxicity of most of the tested benchmark substance and its ease of implementation in a high throughput format, the 53BP1 assay could be proposed as a complementary high-throughput screening genotoxicity assay, in the context of the development of New Approach Methodologies.
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U2 - 10.1371/journal.pone.0288737
DO - 10.1371/journal.pone.0288737
M3 - Article
C2 - 37713377
AN - SCOPUS:85171417845
SN - 1932-6203
VL - 18
JO - PLoS ONE
JF - PLoS ONE
IS - 9 SEPTEMBER
M1 - e0288737
ER -