Overlooked and valuable facts to know in the NRF2/KEAP1 field

Kopacz, Aleksandra; Rojo, Ana I.; Patibandla, Chinmai; Lastra-Martínez, Diego; Piechota-Polańczyk, Aleksandra; Klóska, Damian; Józkowicz, Alicja; Sutherland, Calum; Cuadrado, Antonio; Grochot-Przęczek, Anna

doi:10.1016/j.freeradbiomed.2022.08.044

Cited by 21 publications

(16 citation statements)

References 125 publications

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“…Two bands were detected in the Co cell line (corresponding to around 70 and 90 kDa), whereas only one (70 kDa) band was seen in the Co-dN cell line. Only the 90 kDa band is speci c to the Nrf2 protein as referred by Kopacz et al 11 , the 70 kDa band corresponds to a non-speci c band, as it is also present with the b-actin antibody (Supplementary Fig. S2).…”

Section: Resultsmentioning

confidence: 75%

Characterization of murine isogenic normal and NRF2-KO colon epithelial cells to explore the food contaminants toxicity and oxidative stress involvement.

Dupuy

Fouché

Noirot

et al. 2023

Preprint

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Cell lines are a useful tool for cellular metabolism and xenobiotic toxicity studies, but for modeling biological effects of molecules on healthy cells or cancer promotion it appeared necessary to develop a cellular model in a normal genetic context without mutations inherent to carcinogenic transformations. Colon mucosa is currently the target of xenobiotics such as food contaminants or naturally produced biomolecules from digestion, compelling normal cells to deal with toxic effects that can lead to genotoxicity and carcinogenic transformations. The toxicological properties of such compounds may rely on reactive oxygen species generation (ROS) which causes oxidative stress. One of the major regulators of ROS metabolism and antioxidant cellular defense is the transcription factor Nrf2. We developed a dual-cell model comparing normal murine epithelial cells with their Nrf2-KO isogenic cells generated through the CRISPR /Cas9 technique. HNE (4-hydroxy-2-nonenal), a lipid peroxidation product resulting from red meat digestion, inducing oxidative stress was used as a model molecule to evaluate normal epithelial cell response to food contaminant toxicity, and the importance of Nrf2 in the cellular protection against ROS injury. Implications of glutathione levels, gene regulation, viability, cell proliferation, and genotoxicity are discussed.

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Section: Resultsmentioning

confidence: 75%

Characterization of murine isogenic normal and NRF2-KO colon epithelial cells to explore the food contaminants toxicity and oxidative stress involvement.

Dupuy

Fouché

Noirot

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…In the present work, these curves were obtained for the Nrf2 protein according to Western blot data. We chose the 64 kDa band to obtain the Nrf2 melting curve, since among all the cell lines studied by us, the band above 64 kDa is observed only for the MEF cell line, and, according to the literature data, the 57–68 kDa band corresponds to the Nrf2 fragment, and its intensity changes upon activation of the Nrf2 system [ 37 , 38 , 39 ]. Another band observed at 28 kDa also seems to correspond to the Nrf2 fragment [ 40 ].…”

Section: Resultsmentioning

confidence: 99%

An Approach to Evaluate the Effective Cytoplasmic Concentration of Bioactive Agents Interacting with a Selected Intracellular Target Protein

et al. 2023

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To compare the effectiveness of various bioactive agents reversibly acting within a cell on a target intracellular macromolecule, it is necessary to assess effective cytoplasmic concentrations of the delivered bioactive agents. In this work, based on a simple equilibrium model and the cellular thermal shift assay (CETSA), an approach is proposed to assess effective concentrations of both a delivered bioactive agent and a target protein. This approach was tested by evaluating the average concentrations of nuclear factor erythroid 2-related factor 2 (Nrf2) and Kelch-like ECH-associated-protein 1 (Keap1) proteins in the cytoplasm for five different cell lines (Hepa1, MEF, RAW264.7, 3LL, and AML12) and comparing the results with known literature data. The proposed approach makes it possible to analyze both binary interactions and ternary competition systems; thus, it can have a wide application for the analysis of protein–protein or molecule–protein interactions in the cell. The concentrations of Nrf2 and Keap1 in the cell can be useful not only in analyzing the conditions for the activation of the Nrf2 system, but also for comparing the effectiveness of various drug delivery systems, where the delivered molecule is able to interact with Keap1.

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“…Increasing ROS levels or exposure to electrophilic activators such as sulforaphane, dimethyl fumarate, curcumin, resveratrol, triterpenoids, and quercetin, among many other molecules, modify KEAP1 cysteine residues and can thereby disable NRF2 binding. De novo translated NRF2 translocates to the nucleus [ [126] , [127] , [128] , [129] ] where it heterodimerizes with the small MAF (sMAF) proteins, and this protein complex binds to the NRF2 consensus sequence, the ARE, located in the promoter region of target genes [ 130 ] ( Fig. 2 B).…”

Section: Pharmacological Modulation Of Hif-1α and Nrf2/keap1 Pathways...mentioning

confidence: 99%

“…According to these findings, NRF2 activators may limit the progression of IBD in the early stages by inhibiting inflammation and oxidative stress. However, it should be noted that NRF2 activators may also exert nonspecific effects related to their off-targets [ 128 ].…”

Section: Pharmacological Modulation Of Hif-1α and Nrf2/keap1 Pathways...mentioning

confidence: 99%

Personalized redox medicine in inflammatory bowel diseases: an emerging role for HIF-1α and NRF2 as therapeutic targets

et al. 2023

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Overlooked and valuable facts to know in the NRF2/KEAP1 field

Cited by 21 publications

References 125 publications

Characterization of murine isogenic normal and NRF2-KO colon epithelial cells to explore the food contaminants toxicity and oxidative stress involvement.

Characterization of murine isogenic normal and NRF2-KO colon epithelial cells to explore the food contaminants toxicity and oxidative stress involvement.

An Approach to Evaluate the Effective Cytoplasmic Concentration of Bioactive Agents Interacting with a Selected Intracellular Target Protein

Personalized redox medicine in inflammatory bowel diseases: an emerging role for HIF-1α and NRF2 as therapeutic targets

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