In Vitro Cell Death Determination for Drug Discovery: A Landscape Review of Real Issues

Méry, Benoîte; Guy, Jean‐Baptiste; Vallard, Alexis; Espenel, Sophie; Ardail, Dominique; Rodriguez-Lafrasse, Claire; Rancoule, C.; Magné, Nicolas

doi:10.1177/1179670717691251

Cited by 77 publications

(59 citation statements)

References 56 publications

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“…However, the lack of correlation could also be due to different assay outcomes (cell permeability versus mitochondrial function) and both assays have diagnostic weaknesses. As a vital dye, YO‐PRO is considered a conclusive test for cell death . However, false negatives can occur with cell death that does not involve increased membrane permeability and false positives are possible with high expression of transporters for large cations .…”

Section: Discussionmentioning

confidence: 99%

“…However, false negatives can occur with cell death that does not involve increased membrane permeability and false positives are possible with high expression of transporters for large cations . In the MTT assay, impaired mitochondrial reductase function in otherwise healthy cells will yield falsely low viability results . These factors may lead to discordant results between YO‐PRO and MTT assays …”

Section: Discussionmentioning

confidence: 99%

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RNA expression profiling in sulfamethoxazole‐treated patients with a range of in vitro lymphocyte cytotoxicity phenotypes

Reinhart

Rose

Panyard

et al. 2018

Pharmacology Res & Perspec

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The lymphocyte toxicity assay (LTA) is a proposed surrogate marker of sulfonamide antibiotic hypersensitivity. In the LTA, peripheral blood mononuclear cells (PBMCs) undergo apoptosis more readily in hypersensitive versus tolerant patients when exposed to drug‐hydroxylamine metabolites in vitro. The purpose of this study was to identify key gene transcripts associated with increased cytotoxicity from sulfamethoxazole‐hydroxylamine in human PBMCs in the LTA. The LTA was performed on PBMCs of 10 patients hypersensitive to trimethoprim‐sulfamethoxazole (HS) and 10 drug‐tolerant controls (TOL), using two cytotoxicity assays: YO‐PRO (n = 20) and MTT (n = 12). mRNA expression profiles of PBMCs, enriched for CD8+ T cells, were compared between HS and TOL patients. Transcript expression was interrogated for correlation with % cytotoxicity from YO‐PRO and MTT assays. Correlated transcripts of interest were validated by qPCR. LTA results were not significantly different between HS and TOL patients, and no transcripts were found to be differentially expressed between the two groups. 96 transcripts were correlated with cytotoxicity by YO‐PRO (r = ±.63‐.75, FDR 0.188). Transcripts were selected for validation based on mechanistic plausibility and three were significantly over‐expressed by qPCR in high cytotoxicity patients: multi‐specific organic anion transporter C (ABCC5), mitoferrin‐1 (SLC25A37), and Porimin (TMEM123). These data identify novel transcripts that could contribute to sulfonamide‐hydroxylamine induced cytotoxicity. These include SLC25A37, encoding a mitochondrial iron transporter, ABCC5, encoding an arylamine drug transporter, and TMEM123, encoding a transmembrane protein that mediates cell death.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

RNA expression profiling in sulfamethoxazole‐treated patients with a range of in vitro lymphocyte cytotoxicity phenotypes

Reinhart

Rose

Panyard

et al. 2018

Pharmacology Res & Perspec

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“…For drug discovery, cell death related testing is paramount in oncology because resistance to dying is a hallmark of cancer cells (40). Knowing the cytotoxic activity demonstrated by ELPA, we decided to identify the mechanism of the cytotoxicity induced by the extract.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of the cytotoxic potential of extracts from the genus Passiflora cultived in Brazil against cancer cells

Svf

Âr

et al. 2018

Preprint

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Abstract20 This work aimed to evaluate the cytotoxic potential against cancer cells of Passiflora genus 21 plant species cultivated in Brazil and identify the mechanism of cytotoxicity induced by the 22 most promising extract. Leaf extracts from 14 Passiflora (P.) species were obtained ASE and 23 in vitro cytotoxicity evaluated against cancer cell lines using MTT assay at a single . CC-BY 4.0 International license It is made available under a (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.

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“…In in vitro toxicity evaluation, choosing the right cytotoxicity assay, or a combination of different ones is not trivial and needs to be chosen wisely (Méry et al, 2017). It is recommended to use a larger variety of measures for the endpoint cell death because cells can die by different modes, which are sometimes difficult to distinguish, and it is thereby essential that the measurement of cell death involves complementary methodologies.…”

Section: Chicken and Xenopusmentioning

confidence: 99%

Literature review and appraisal on alternative neurotoxicity testing methods

Masjosthusmann

Barenys

El‐Gamal

et al. 2018

EFSA Supporting Publications

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The goal of this review was the evaluation of information on assessment methods in the field of alternative neurotoxicity (NT) testing. We therefore performed a systematic and comprehensive collection of scientific literature (in English) from the past 27 years until mid of 2017 on state of the art alternative testing methods including in vitro test methods, in silico methods and alternative non-mammalian models. This review identified a variety of test methods that have the ability to predict NT of chemicals based on predefined key NT endpoint categories (27). Those endpoint categories were derived from the Mode of Action (MoA) of known human neurotoxicants. Pre-evaluated MoAs of human neurotoxicants allowed the identification of performance characteristics with regard to the ability of a test system to correctly predict a chemical effect on an endpoint category. The most predictive in vitro model that covers a large variety of endpoint categories are primary rodent cells or tissues. Human based systems derived from induced pluripotent stem cells (iPSC) are promising and warrant human relevance. There is however not yet sufficient data on these models to demonstrate their suitability to reliably substitute primary rodent cells for NT testing purposes. Test methods for glia toxicity are rare and glia endpoint categories are clearly underrepresented. Therefore, a focus for future method development should be placed on glia, astrocytes, oligodendrocytes and microglia based models, preferably in a co-culture se up. The review on in silico methods, resulted into 54 QSARs publications, relevant for NT, of which 39 on blood brain barrier (BBB) permeation. The QSARs available in the publications were developed from data on drugs and chemicals, but there appears a limited set of experimental data for chemicals and pesticides on blood-brain barrier passage. The evaluation of NT methods using alternative whole organism approaches demonstrated a majority of data for C. elegans (nematode species), represented with high true prediction (96%). The main endpoint category was inhibition of cholinergic transmission, with specific endpoints for AChE activity and motor activity, the latter confirming the added value of a whole organism approach among alternative models. Though D. rerio, the zebrafish model appeared a www.efsa.europa.eu/publications 2 EFSA Supporting publication 2018:EN-1410The present document has been produced and adopted by the bodies identified above as authors. This task has been carried out exclusively by the authors in the context of a contract between the European Food Safety Authority and the authors, awarded following a tender procedure. The present document is published complying with the transparency principle to which the Authority is subject. It may not be considered as an output adopted by the Authority. The European Food Safety Authority reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights ...

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In Vitro Cell Death Determination for Drug Discovery: A Landscape Review of Real Issues

Cited by 77 publications

References 56 publications

RNA expression profiling in sulfamethoxazole‐treated patients with a range of in vitro lymphocyte cytotoxicity phenotypes

RNA expression profiling in sulfamethoxazole‐treated patients with a range of in vitro lymphocyte cytotoxicity phenotypes

Evaluation of the cytotoxic potential of extracts from the genus Passiflora cultived in Brazil against cancer cells

Literature review and appraisal on alternative neurotoxicity testing methods

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