VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines
Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOX) involvement has been established in the oncogenic cell signaling of acute myeloid leukemia (AML) cells and in the crosstalk with their niche. We have shown an expression of NOX subunits in AML cell lines while NOX activity is lacking...
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MDPI AG
2020-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/15/5470 |
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| author | Maya El Dor Hassan Dakik Marion Polomski Eloi Haudebourg Marie Brachet Fabrice Gouilleux Gildas Prié Kazem Zibara Frédéric Mazurier |
| author_facet | Maya El Dor Hassan Dakik Marion Polomski Eloi Haudebourg Marie Brachet Fabrice Gouilleux Gildas Prié Kazem Zibara Frédéric Mazurier |
| author_sort | Maya El Dor |
| collection | DOAJ |
| description | Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOX) involvement has been established in the oncogenic cell signaling of acute myeloid leukemia (AML) cells and in the crosstalk with their niche. We have shown an expression of NOX subunits in AML cell lines while NOX activity is lacking in the absence of exogenous stimulation. Here, we used AML cell lines as models to investigate the specificity of VAS3947, a current NOX inhibitor. Results demonstrated that VAS3947 induces apoptosis in AML cells independently of its anti-NOX activity. High-performance liquid chromatography (HPLC) and mass spectrometry analyses revealed that VAS3947 thiol alkylates cysteine residues of glutathione (GSH), while also interacting with proteins. Remarkably, VAS3947 decreased detectable GSH in the MV-4-11 cell line, thereby suggesting possible oxidative stress induction. However, a decrease in both cytoplasmic and mitochondrial reactive oxygen species (ROS) levels was observed by flow cytometry without disturbance of mitochondrial mass and membrane potential. Thus, assuming the consequences of VAS3947 treatment on protein structure, we examined its impact on endoplasmic reticulum (ER) stress. An acute unfolded protein response (UPR) was triggered shortly after VAS3947 exposure, through the activation of inositol-requiring enzyme 1α (IRE1α) and PKR-like endoplasmic reticulum kinase (PERK) pathways. Overall, VAS3947 induces apoptosis independently of anti-NOX activity, via UPR activation, mainly due to aggregation and misfolding of proteins. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T18:04:08Z |
| publishDate | 2020-07-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-58beb1b2f2f04826b895907e1feabc392023-11-20T08:35:55ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-07-012115547010.3390/ijms21155470VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell LinesMaya El Dor0Hassan Dakik1Marion Polomski2Eloi Haudebourg3Marie Brachet4Fabrice Gouilleux5Gildas Prié6Kazem Zibara7Frédéric Mazurier8EA 7501 GICC, University of Tours, CNRS ERL7001 LNOx, Bâtiment Dutrochet, 10 boulevard Tonnellé, BP3223, CEDEX 1, 37032 Tours, FranceEA 7501 GICC, University of Tours, CNRS ERL7001 LNOx, Bâtiment Dutrochet, 10 boulevard Tonnellé, BP3223, CEDEX 1, 37032 Tours, FranceEA 7501 GICC, University of Tours, IMT, 31 Avenue Monge, 37200 Tours, FranceEA 7501 GICC, University of Tours, IMT, 31 Avenue Monge, 37200 Tours, FranceEA 7501 GICC, University of Tours, CNRS ERL7001 LNOx, Bâtiment Dutrochet, 10 boulevard Tonnellé, BP3223, CEDEX 1, 37032 Tours, FranceEA 7501 GICC, University of Tours, CNRS ERL7001 LNOx, Bâtiment Dutrochet, 10 boulevard Tonnellé, BP3223, CEDEX 1, 37032 Tours, FranceEA 7501 GICC, University of Tours, IMT, 31 Avenue Monge, 37200 Tours, FrancePRASE, Beirut, LebanonEA 7501 GICC, University of Tours, CNRS ERL7001 LNOx, Bâtiment Dutrochet, 10 boulevard Tonnellé, BP3223, CEDEX 1, 37032 Tours, FranceNicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOX) involvement has been established in the oncogenic cell signaling of acute myeloid leukemia (AML) cells and in the crosstalk with their niche. We have shown an expression of NOX subunits in AML cell lines while NOX activity is lacking in the absence of exogenous stimulation. Here, we used AML cell lines as models to investigate the specificity of VAS3947, a current NOX inhibitor. Results demonstrated that VAS3947 induces apoptosis in AML cells independently of its anti-NOX activity. High-performance liquid chromatography (HPLC) and mass spectrometry analyses revealed that VAS3947 thiol alkylates cysteine residues of glutathione (GSH), while also interacting with proteins. Remarkably, VAS3947 decreased detectable GSH in the MV-4-11 cell line, thereby suggesting possible oxidative stress induction. However, a decrease in both cytoplasmic and mitochondrial reactive oxygen species (ROS) levels was observed by flow cytometry without disturbance of mitochondrial mass and membrane potential. Thus, assuming the consequences of VAS3947 treatment on protein structure, we examined its impact on endoplasmic reticulum (ER) stress. An acute unfolded protein response (UPR) was triggered shortly after VAS3947 exposure, through the activation of inositol-requiring enzyme 1α (IRE1α) and PKR-like endoplasmic reticulum kinase (PERK) pathways. Overall, VAS3947 induces apoptosis independently of anti-NOX activity, via UPR activation, mainly due to aggregation and misfolding of proteins.https://www.mdpi.com/1422-0067/21/15/5470VAS3947NADPH oxidasesleukemiacysteine thiol alkylationoxidative stressendoplasmic reticulum |
| spellingShingle | Maya El Dor Hassan Dakik Marion Polomski Eloi Haudebourg Marie Brachet Fabrice Gouilleux Gildas Prié Kazem Zibara Frédéric Mazurier VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines International Journal of Molecular Sciences VAS3947 NADPH oxidases leukemia cysteine thiol alkylation oxidative stress endoplasmic reticulum |
| title | VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines |
| title_full | VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines |
| title_fullStr | VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines |
| title_full_unstemmed | VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines |
| title_short | VAS3947 Induces UPR-Mediated Apoptosis through Cysteine Thiol Alkylation in AML Cell Lines |
| title_sort | vas3947 induces upr mediated apoptosis through cysteine thiol alkylation in aml cell lines |
| topic | VAS3947 NADPH oxidases leukemia cysteine thiol alkylation oxidative stress endoplasmic reticulum |
| url | https://www.mdpi.com/1422-0067/21/15/5470 |
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