Cigarette smoke extract induced exosome release is mediated by depletion of exofacial thiols and can be inhibited by thiol-antioxidants

Benedikter, Birke J.; Volgers, Charlotte; Eijck, Pascalle H. van; Wouters, Emiel F.M.; Savelkoul, Paul H. M.; Reynaert, Niki L.; Haenen, Guido R.M.M.; Rohde, Gernot; Weseler, Antje R.; Stassen, Frank R. M.

doi:10.1016/j.freeradbiomed.2017.03.026

Cited by 66 publications

(84 citation statements)

References 60 publications

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“…Cell culture and exposure BEAS-2B airway epithelial cells (ATCC CRL-9609) were cultured in RPMI 1640 with 10% (v/v) foetal calf serum (FCS, Lonza) and all cell culture flasks or plates were precoated with fibronectin as described previously [5]. For flow cytometric analysis of EVs or cells, 2 × 10 5 cells per well were seeded on a 12-well plate (Costar) and for EV-isolations, 4 × 10 6 cells were seeded per T75 (Costar) and allowed to attach overnight.…”

Section: Methodsmentioning

confidence: 99%

“…All antibodies were purchased from BD Biosciences. EVs were detected by bead-based flow cytometry as described previously with some adaptations [5]. Beads (3.5 × 10 8 /ml; 4 μm aldehyde/sulphate latex beads; Thermo Fisher Scientific) were coated with 0.125 mg/ml of mouse antihuman CD63 antibody (Clone H5C6), anti-human CD81 (clone JS-81), anti-human CD9 (Clone M-L13), or an equal mixture of all three antibodies by overnight incubation in MES buffer on a shaker at 6500 rpm.…”

Section: Ev Detection Using Bead-based Flow Cytometrymentioning

confidence: 99%

“…Although the airway epithelium forms the first line of contact with inhaled cigarette smoke, studies on the functions of EVs released by smoke-exposed airway epithelial cells are scarce. Previously, we have shown that airway epithelial cells secrete small EVs (mode size 110 nm) expressing the tetraspanins CD63, CD81 and CD9, at control conditions and when exposed to CSE [5,13]. In this study, we aimed to predict the functions of these EVs.…”

Section: Introductionmentioning

confidence: 99%

“…Tobacco consumption forms an important risk factor for several chronic diseases including lung cancer, chronic obstructive pulmonary disease (COPD) and cardiovascular disease [1]. We and others have shown that cigarette smoke extract (CSE) causes increased release of extracellular vesicles (EVs) by different cell types in vitro [2][3][4][5]. Moreover, smokers have increased concentrations of circulating EVs in vivo [4,6,7].…”

Section: Introductionmentioning

confidence: 99%

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Proteomic analysis reveals procoagulant properties of cigarette smoke‐induced extracellular vesicles

Benedikter

Bouwman

Heinzmann

et al. 2019

J of Extracellular Vesicle

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Airway epithelial cells secrete extracellular vesicles (EVs) under basal conditions and when exposed to cigarette smoke extract (CSE). Getting insights into the composition of these EVs will help unravel their functions in homeostasis and smoking-induced pathology. Here, we characterized the proteomic composition of basal and CSE-induced airway epithelial EVs. BEAS-2B cells were left unexposed or exposed to 1% CSE for 24 h, followed by EV isolation using ultrafiltration and size exclusion chromatography. Isolated EVs were labelled with tandem mass tags and their proteomic composition was determined using nano-LC-MS/MS. Tissue factor (TF) activity was determined by a factor Xa generation assay, phosphatidylserine (PS) content by prothrombinase assay and thrombin generation using calibrated automated thrombogram (CAT). Nano-LC-MS/MS identified 585 EV-associated proteins with high confidence. Of these, 201 were differentially expressed in the CSE-EVs according to the moderated t-test, followed by false discovery rate (FDR) adjustment with the FDR threshold set to 0.1. Functional enrichment analysis revealed that 24 proteins of the pathway haemostasis were significantly up-regulated in CSE-EVs, including TF. Increased TF expression on CSE-EVs was confirmed by bead-based flow cytometry and was associated with increased TF activity. CSE-EVs caused faster and more thrombin generation in normal human plasma than control-EVs, which was partly TF-, but also PS-dependent. In conclusion, proteomic analysis allowed us to predict procoagulant properties of CSE-EVs which were confirmed in vitro. Cigarette smoke-induced EVs may contribute to the increased cardiovascular and respiratory risk observed in smokers. ARTICLE HISTORY

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Section: Methodsmentioning

confidence: 99%

Section: Ev Detection Using Bead-based Flow Cytometrymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Proteomic analysis reveals procoagulant properties of cigarette smoke‐induced extracellular vesicles

Benedikter

Bouwman

Heinzmann

et al. 2019

J of Extracellular Vesicle

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“…CSE was prepared according to previously described method (Slebos et al, 2007;Benedikter et al, 2017). Briefly, 3R4F Research Cigarettes (University of Kentucky, USA) were smoked using a peristaltic pump machine (Fisher Scientific, Pittsburgh, PA, USA).…”

Section: Cse Preparationmentioning

confidence: 99%

Overexpression of MECP2 attenuates cigarette smoke extracts induced lung epithelial cell injury by promoting CYP1B1 methylation

et al. 2020

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MECP2 (Methyl-CpG-binding protein 2) has been shown to have a critical role in regulating DNA methylation against smoke exposed lung injury. However, the biological function of MECP2 and the underlying molecular mechanism remains elusive. Human bronchial epithelial (16HBE) and alveolar type II epithelial cells (AECII) were exposed to increasing concentrations of cigarette smoke extracts (CSE) solution to establish CSE-induced lung epithelial cell injury models. Our findings revealed that MECP2 was down-regulated, while CYP1B1 was up-regulated in CSE-induced lung epithelial cell injury models by quantitative real time PCR, western blotting and immunofluorescence staining. Down-regulated CYP1B1 was ascribed to the demethylation of its promoter by methylation-specific PCR (MSP). The in vitro experiments further showed that MECP2 overexpression significantly attenuated CSE-triggered cell growth attenuation, cell cycle arrest, apoptosis and ROS generation in lung epithelial cells by CCK-8 and flow cytometry assays. In molecular level, we further demonstrated that MECP2 overexpression obviously suppressed the expression of CYP1B1 through enhancing DNA methylation. Therefore, our data suggest that MECP2 protects against CSE-induced lung epithelial cell injury possibly through down-regulating CYP1B1 expression via elevating its methylation status.

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Extracellular vesicles in toxicological studies: key roles in communication between environmental stress and adverse outcomes

Qin

Huang

2020

J of Applied Toxicology

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External stressors, especially environmental toxicants can disturb biological homeostasis and thus lead to adverse health effects. However, there is limited understanding of how cells directly exposed to stressors transmit the signals to cells indirectly in contact with stressors. Extracellular vesicles (EVs) are receiving increasing attention as signal transductors between various types of cells in organisms. Cargo in EVs, including RNAs, proteins, lipids, and other signal molecules can be transferred between cells and become critical determining factors of intercellular communication. EVs can be a powerful mediator of environmental stimuli. It has been shown that external stressors reshape the secretion of EVs, modify the composition of EVs, and thus influence the mediating function of EVs. These abnormal EVs can lead to dysfunction of recipient cells, and even the pathogenesis of diseases. In this review, we first summarized current knowledge about the responses of EVs to external stimuli, including chemicals and chemical mixtures. Then we explained how these altered EVs regulate signal pathways in recipient cells, thus mediating physio‐pathological responses in detail. The most up‐to‐date evidence from molecular, cellular, animal and human levels was synthesized to systematically address the mediating roles of EVs. EVs can be regarded as a bridge to link external stressors and internal response. Further toxicological and molecular epidemiological studies are expected to provide further insight into the roles of EVs in toxicology. The gaps in the engulfment of toxicants into EVs are listed as the priority to be solved in future studies.

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Cigarette smoke extract induced exosome release is mediated by depletion of exofacial thiols and can be inhibited by thiol-antioxidants

Cited by 66 publications

References 60 publications

Proteomic analysis reveals procoagulant properties of cigarette smoke‐induced extracellular vesicles

Proteomic analysis reveals procoagulant properties of cigarette smoke‐induced extracellular vesicles

Overexpression of MECP2 attenuates cigarette smoke extracts induced lung epithelial cell injury by promoting CYP1B1 methylation

Extracellular vesicles in toxicological studies: key roles in communication between environmental stress and adverse outcomes

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