A cell-based biosensor system HepG2CDKN1A–DsRed for rapid and simple detection of genotoxic agents

Blagus, Tanja; Žager, Valerija; Čemažar, Maja; Serša, Gregor; Kamenšek, Urška; Žegura, Bojana; Nunić, Jana; Filipič, Metka

doi:10.1016/j.bios.2014.05.002

Cited by 14 publications

(7 citation statements)

References 50 publications

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“…With regard to in vitro studies, various assays have been performed ( Table 5 ) [ 26 , 27 , 28 , 35 , 53 , 60 , 63 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 ], mainly in mammalian systems, which demonstrated the pro-genotoxic activity of CYN.…”

Section: Toxicity Mechanismsmentioning

confidence: 99%

In Vitro Toxicological Assessment of Cylindrospermopsin: A Review

Pichardo

Cameán

Jos

2017

Toxins

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Cylindrospermopsin (CYN) is a cyanobacterial toxin that is gaining importance, owing to its increasing expansion worldwide and the increased frequency of its blooms. CYN mainly targets the liver, but also involves other organs. Various mechanisms have been associated with its toxicity, such as protein synthesis inhibition, oxidative stress, etc. However, its toxic effects are not yet fully elucidated and additional data for hazard characterization purposes are required. In this regard, in vitro methods can play an important role, owing to their advantages in comparison to in vivo trials. The aim of this work was to compile and evaluate the in vitro data dealing with CYN available in the scientific literature, focusing on its toxicokinetics and its main toxicity mechanisms. This analysis would be useful to identify research needs and data gaps in order to complete knowledge about the toxicity profile of CYN. For example, it has been shown that research on various aspects, such as new emerging toxicity effects, the toxicity of analogs, or the potential interaction of CYN with other cyanotoxins, among others, is still very scarce. New in vitro studies are therefore welcome.

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Section: Toxicity Mechanismsmentioning

confidence: 99%

In Vitro Toxicological Assessment of Cylindrospermopsin: A Review

Pichardo

Cameán

Jos

2017

Toxins

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“…More recently, analogous systems have been developed in yeast and mammalian cells − that should more accurately reflect toxicological responses in humans. In one of the key examples, the promoter and regulatory sequences of a g rowth a rrest and D NA d amage gene, GADD45a, the expression of which is upregulated in human cells in response to DNA damage, were coupled to the expression of GFP in a human lymphoblastoid TK6 cell line .…”

Section: Genetically Encoded Response Circuits: Augmented Toxicologic...mentioning

confidence: 99%

“…GreenScreen HC also compares well to alternative genotoxicity tests such as the SOS-Chromotest and Mini Ames II. − This has been the forerunner of a number of similar reporter gene assays based on linking the regulatory sequences for some protein involved in a response to toxic insult (multiple mechanisms of genotoxicity, oxidative stress, etc.) to a fluorescent or luminescent reporter gene. ,− These have been reviewed recently elsewhere and will not be covered in detail here. Again, however, most lack any endogenous bioactivation system to enable the detection of toxicity due to metabolites rather than parent compounds.…”

Section: Genetically Encoded Response Circuits: Augmented Toxicologic...mentioning

confidence: 99%

Prospects for Applying Synthetic Biology to Toxicology: Future Opportunities and Current Limitations for the Repurposing of Cytochrome P450 Systems

Behrendorff

Gillam

2016

Chem. Res. Toxicol.

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The 30 years since the inception of Chemical Research in Toxicology, game-changing advances in chemical and molecular biology, the fundamental disciplines underpinning molecular toxicology, have been made. While these have led to important advances in the study of mechanisms by which chemicals damage cells and systems, there has been less focus on applying these advances to prediction, detection, and mitigation of toxicity. Over the last ∼15 years, synthetic biology, the repurposing of biological "parts" in systems engineered for useful ends, has been explored in other areas of the biomedical and life sciences, for such applications as detecting metabolites, drug discovery and delivery, investigating disease mechanisms, improving medical treatment, and producing useful chemicals. These examples provide models for the application of synthetic biology to toxicology, which, for the most part, has not yet benefited from such approaches. In this perspective, we review the synthetic biology approaches that have been applied to date and speculate on possible short to medium term and "blue sky" aspirations for synthetic biology, particularly in clinical and environmental toxicology. Finally, we point out key hurdles that must be overcome for the full potential of synthetic biology to be realized.

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“…To overcome these problems, rapid high-throughput cell-based assays that measure cell responses and gene induction are being developed to provide early warning on metabolic pathway activations and toxicity [4]. Among these assays, whole cell biosensors in HepaRG [21,26,27] and HepG2 [26,27,28,29] hepatoma cell lines have been developed in order to detect mitochondrial dysfunction, toxicity, metabolism and genotoxicity [4].…”

Section: Introductionmentioning

confidence: 99%

Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity

Vlach¹,

Quesnot²,

Dubois-Pot-Schneider

et al. 2019

Sensors

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Human hepatoma HepaRG cells express most drug metabolizing enzymes and constitute a pertinent in vitro alternative cell system to primary cultures of human hepatocytes in order to determine drug metabolism and evaluate the toxicity of xenobiotics. In this work, we established novel transgenic HepaRG cells transduced with lentiviruses encoding the reporter green fluorescent protein (GFP) transcriptionally regulated by promoter sequences of cytochromes P450 (CYP) 1A1/2, 2B6 and 3A4 genes. Here, we demonstrated that GFP-biosensor transgenes shared similar expression patterns with the corresponding endogenous CYP genes during proliferation and differentiation in HepaRG cells. Interestingly, differentiated hepatocyte-like HepaRG cells expressed GFP at higher levels than cholangiocyte-like cells. Despite weaker inductions of GFP expression compared to the strong increases in mRNA levels of endogenous genes, we also demonstrated that the biosensor transgenes were induced by prototypical drug inducers benzo(a)pyrene and phenobarbital. In addition, we used the differentiated biosensor HepaRG cells to evidence that pesticide mancozeb triggered selective cytotoxicity of hepatocyte-like cells. Our data demonstrate that these new biosensor HepaRG cells have potential applications in the field of chemicals safety evaluation and the assessment of drug hepatotoxicity.

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A cell-based biosensor system HepG2CDKN1A–DsRed for rapid and simple detection of genotoxic agents

Cited by 14 publications

References 50 publications

In Vitro Toxicological Assessment of Cylindrospermopsin: A Review

In Vitro Toxicological Assessment of Cylindrospermopsin: A Review

Prospects for Applying Synthetic Biology to Toxicology: Future Opportunities and Current Limitations for the Repurposing of Cytochrome P450 Systems

Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity

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