Development of a panel of high-throughput reporter-gene assays to detect genotoxicity and oxidative stress

Linden, Sander C. van der; Bergh, Anne R. M. von; Vught-Lussenburg, Barbara M.A. van; Jonker, Lydia; Teunis, Marc; Krul, Cyrille; Burg, Bart van der

doi:10.1016/j.mrgentox.2013.09.009

Cited by 105 publications

(79 citation statements)

References 38 publications

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“…The existing data are strongest for PD, ALS, and MS, but ongoing experiments in AD and HD should provide more insight into how important Nrf2 is in these diseases as well. Numerous cell-based and in silico high-throughput screens have identified novel Nrf2-activating compounds [199–205]. The most promising approach may be to find compounds that noncovalently disrupt the DLG or ETGE motif interactions with Keap1 [206, 207].…”

Section: Discussionmentioning

confidence: 99%

Nrf2—a therapeutic target for the treatment of neurodegenerative diseases

Johnson

2015

Free Radical Biology and Medicine

278

228

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The brain is very sensitive to changes in redox status; thus maintaining redox homeostasis in the brain is critical for the prevention of accumulating oxidative damage. Aging is the primary risk factor for developing neurodegenerative diseases. In addition to age, genetic and environmental risk factors have also been associated with disease development. The primary reactive insults associated with the aging process are a result of oxidative stress (OS) and nitrosative stress (NS). Markers of increased oxidative stress, protein and DNA modification, inflammation, and dysfunctional proteostasis have all been implicated in contributing to the progression of neurodegeneration. The ability of the cell to combat OS/NS and maintain a clearance mechanism for misfolded aggregating proteins determines whether or not it will survive. A critical pathway in this regard is the Nrf2 (nuclear factor erythroid 2-related factor 2)- antioxidant response element (ARE) pathway. Nrf2 activation has been shown to mitigate a number of pathologic mechanisms associated with Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, and multiple sclerosis. This review will focus on the role of Nrf2 in these diseases and the potential for Nrf2 activation to mitigate disease progression.

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Section: Discussionmentioning

confidence: 99%

Nrf2—a therapeutic target for the treatment of neurodegenerative diseases

Johnson

2015

Free Radical Biology and Medicine

278

228

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“…Likewise, inclusion of metabolic steps can contribute to improved predictions at least at the level of the individual assays, as has been shown recently for steroid-receptor mediated responses [9,72], or genotoxicity [60]. Our current study focused on hazard identification primarily and did not include these metabolic and pharmacokinetic steps which will likely lead to further improvements of predictions.…”

Section: Metabolism/kineticsmentioning

confidence: 97%

“…This screening panel was completed with a range of U2-OS-based singly transfected lines expressing a reporter gene only, which are designed to selectively measure the activity of main intracellular signaling pathways. This included assays to assess transcriptional activation by the p53 protein; p53 CALUX line, transcriptional activation of the cyclin-dependent kinase inhibitor p21 promoter region; p21 CALUX, the oxidative stress responsive nrf2 pathway; Nrf2 CALUX [60], the endoplasmic reticulum stress pathway ESRE CALUX, The Wnt signaling pathway; TCF CALUX, the activator protein 1 pathway; AP1 CALUX, the nuclear factor B pathway; NFB CALUX and the hypoxia-inducible factor induced pathway; HIF1␣ CALUX [11]. In general, tests were carried out in a way to measure agonistic effects of compounds, while in several cases deemed relevant, also tests were carried out in the additional presence of reference agonist at an EC50 level in order to assess antagonism [15].…”

Section: Calux Batterymentioning

confidence: 99%

A high throughput screening system for predicting chemically-induced reproductive organ deformities

Burg

Pieterse

Buist³

et al. 2015

Reproductive Toxicology

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“…These responses can also be relevant in reproductive toxicity, however. A control cell line, called cytotox CALUX has been generated which constitutively expresses the luciferase gene [27]. To validate this approach of using limited sets of selective assays it was shown that a single selective and extensively validated CALUX assay (ERalpha CALUX…”

Section: Establish a High Throughput Mechanistic Pathway Screen For Rmentioning

confidence: 99%

“…This tool was successfully constructed and installed and all CALUX HTS screening data have been uploaded. Finally, the CALUX HTS panel endogenously expresses little metabolic activity but can be run with and without S9 metabolic fractions [27].…”

Section: Establish a High Throughput Mechanistic Pathway Screen For Rmentioning

confidence: 99%

The ChemScreen project to design a pragmatic alternative approach to predict reproductive toxicity of chemicals

Burg

Wedebye

Dietrich

et al. 2015

Reproductive Toxicology

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a b s t r a c tThere is a great need for rapid testing strategies for reproductive toxicity testing, avoiding animal use. The EU Framework program 7 project ChemScreen aimed to fill this gap in a pragmatic manner preferably using validated existing tools and place them in an innovative alternative testing strategy. In our approach we combined knowledge on critical processes affected by reproductive toxicants with knowledge on the mechanistic basis of such effects. We used in silico methods for prescreening chemicals for relevant toxic effects aiming at reduced testing needs. For those chemicals that need testing we have set up an in vitro screening panel that includes mechanistic high throughput methods and lower throughput assays that measure more integrative endpoints. In silico pharmacokinetic modules were developed for rapid exposure predictions via diverse exposure routes. These modules to match in vitro and in vivo exposure levels greatly improved predictivity of the in vitro tests. As a further step, we have generated examples how to predict reproductive toxicity of chemicals using available data. We have executed formal validations of panel constituents and also used more innovative manners to validate the test panel using mechanistic approaches. We are actively engaged in promoting regulatory acceptance of the tools developed as an essential step towards practical application, including case studies for read-across purposes. With this approach, a significant saving in animal use and associated costs seems very feasible.

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Development of a panel of high-throughput reporter-gene assays to detect genotoxicity and oxidative stress

Cited by 105 publications

References 38 publications

Nrf2—a therapeutic target for the treatment of neurodegenerative diseases

Nrf2—a therapeutic target for the treatment of neurodegenerative diseases

A high throughput screening system for predicting chemically-induced reproductive organ deformities

The ChemScreen project to design a pragmatic alternative approach to predict reproductive toxicity of chemicals

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