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Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs) / DEMI PRITCHARD

Swansea University Author: DEMI PRITCHARD

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DOI (Published version): 10.23889/SUthesis.66287

Abstract

This work aimed to develop an in vitro testing battery, to detect non-genotoxic carcinogens (NGCs). Currently, in vitro test systems aim at detecting genotoxic carcinogens (GCs) whereas, the gold standard test for NGCs is the two-year rodent bioassay. There are numerous problems with this method suc...

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Published: Swansea, Wales, UK 2024
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
Supervisor: Pritchard, Demi J. ; Chapman, Katherin E. ; Doak, Shareen H. ; Jenkins, Gareth J. S.
URI: https://cronfa.swan.ac.uk/Record/cronfa66287
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spelling v2 66287 2024-05-03 Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs) 0db60e95c1db88f6507f59402aaedac5 DEMI PRITCHARD DEMI PRITCHARD true false 2024-05-03 This work aimed to develop an in vitro testing battery, to detect non-genotoxic carcinogens (NGCs). Currently, in vitro test systems aim at detecting genotoxic carcinogens (GCs) whereas, the gold standard test for NGCs is the two-year rodent bioassay. There are numerous problems with this method such as long experimental periods, high costs and many ethical concerns. NGCs make up ̴15% of all carcinogens, highlighting the importance of their correct detection and labelling. The in vitro test battery includes a selection of endpoints comprised primarily from the hallmarks of cancer. The endpoints explored were acute and chronic genotoxicity assessment via the mononucleated micronucleus (Mn) assay; cytotoxicity assessment utilising RPD; cell cycle and apoptosis assessment using flow cytometry; gene expression analysis using a PCR array and determining mitochondrial health using the Seahorse analyser. Six carcinogens were assessed using the multi-endpoint, in vitro testing system. These chemicals included nickel chloride (NiCl2), sodium meta arsenite (NaMAr), cacodylic acid (GC- included to compare with NaMAr), rosuvastatin, chloroprene and 2,3,7,8-tetrachlorodibenzodioxin (TCDD). This work showed that some NGCs can behave as GCs so there may be some mislabelling of carcinogens. NaMAr indicated genotoxicity due to a chronic dosing approachand NiCl2 also showed genotoxicity but in an acute setting. The correct categorisation of these carcinogens is of increasing importance due to the use in consumer product for example. The in vitro battery appears to be a promising first step in detecting and understanding the mechanism(s) of action (MoA) used by different NGCs. This work highlighted some potentially mislabelled NGCs, such as NiCl2 and NaMAr, which is an important novel finding. The PCR array proved to be very useful to identify genes of interest, that could help unpick the MoA of each diverse NGC. Next steps could include utilising a targeted three-dimensional (3D) spheroid approach based on the exposure route (E.g. Rosuvastatin is ingested so a gut model). Chronic dosing is an important real-life component of the test battery and is required to improve the whole picture. These additional approaches, together with an in vitro test battery, could improve detection and provide a complete narrative of the MoAs utilized by these complex carcinogens. E-Thesis Swansea, Wales, UK Genetic toxicology, Non-genotoxic carcinogens, cancer, micronucleus assay 5 3 2024 2024-03-05 10.23889/SUthesis.66287 COLLEGE NANME COLLEGE CODE Swansea University Pritchard, Demi J. ; Chapman, Katherin E. ; Doak, Shareen H. ; Jenkins, Gareth J. S. Doctoral Ph.D NC3Rs (JAF1056-100) NC3Rs (JAF1056-100) 2024-05-03T12:41:09.2997617 2024-05-03T10:31:34.8981515 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science DEMI PRITCHARD 1 66287__30260__8d1fdf4cbd7c45a9bb79f7947d4f242f.pdf Pritchard_Demi_J_PhD_Thesis_Final_Redacted_Signature.pdf 2024-05-03T10:56:02.2183738 Output 5369289 application/pdf E-Thesis – open access true Copyright: The Author, Demi Jade Pritchard, 2024. true eng
title Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
spellingShingle Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
DEMI PRITCHARD
title_short Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
title_full Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
title_fullStr Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
title_full_unstemmed Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
title_sort Developing an in vitro repeat-dose approach to detect non-genotoxic carcinogens (NGCs)
author_id_str_mv 0db60e95c1db88f6507f59402aaedac5
author_id_fullname_str_mv 0db60e95c1db88f6507f59402aaedac5_***_DEMI PRITCHARD
author DEMI PRITCHARD
author2 DEMI PRITCHARD
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publishDate 2024
institution Swansea University
doi_str_mv 10.23889/SUthesis.66287
college_str Faculty of Medicine, Health and Life Sciences
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hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
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description This work aimed to develop an in vitro testing battery, to detect non-genotoxic carcinogens (NGCs). Currently, in vitro test systems aim at detecting genotoxic carcinogens (GCs) whereas, the gold standard test for NGCs is the two-year rodent bioassay. There are numerous problems with this method such as long experimental periods, high costs and many ethical concerns. NGCs make up ̴15% of all carcinogens, highlighting the importance of their correct detection and labelling. The in vitro test battery includes a selection of endpoints comprised primarily from the hallmarks of cancer. The endpoints explored were acute and chronic genotoxicity assessment via the mononucleated micronucleus (Mn) assay; cytotoxicity assessment utilising RPD; cell cycle and apoptosis assessment using flow cytometry; gene expression analysis using a PCR array and determining mitochondrial health using the Seahorse analyser. Six carcinogens were assessed using the multi-endpoint, in vitro testing system. These chemicals included nickel chloride (NiCl2), sodium meta arsenite (NaMAr), cacodylic acid (GC- included to compare with NaMAr), rosuvastatin, chloroprene and 2,3,7,8-tetrachlorodibenzodioxin (TCDD). This work showed that some NGCs can behave as GCs so there may be some mislabelling of carcinogens. NaMAr indicated genotoxicity due to a chronic dosing approachand NiCl2 also showed genotoxicity but in an acute setting. The correct categorisation of these carcinogens is of increasing importance due to the use in consumer product for example. The in vitro battery appears to be a promising first step in detecting and understanding the mechanism(s) of action (MoA) used by different NGCs. This work highlighted some potentially mislabelled NGCs, such as NiCl2 and NaMAr, which is an important novel finding. The PCR array proved to be very useful to identify genes of interest, that could help unpick the MoA of each diverse NGC. Next steps could include utilising a targeted three-dimensional (3D) spheroid approach based on the exposure route (E.g. Rosuvastatin is ingested so a gut model). Chronic dosing is an important real-life component of the test battery and is required to improve the whole picture. These additional approaches, together with an in vitro test battery, could improve detection and provide a complete narrative of the MoAs utilized by these complex carcinogens.
published_date 2024-03-05T12:41:09Z
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