DOI: 10.5061/dryad.3n39r6k
title: Data from: Polycyclic aromatic hydrocarbons can trigger hepatocyte release of extracellular vesicles by various mechanisms of action depending on their affinity for the aryl hydrocarbon receptor.

description: Extracellular vesicles (EVs) are membrane enclosed nanostructures released
 by cells into the extracellular environment. As major actors of
 physiological intercellular communication, they have been shown to be
 pathogenic mediators of several liver diseases. EVs also appear to be
 potential actors of drug-induced liver injury, but nothing is known
 concerning environmental pollutants. We aimed to study the impact of
 polycyclic aromatic hydrocarbons (PAHs), major contaminants, on
 hepatocyte-derived EV production, with a special focus on hepatocyte
 death. Three PAHs were selected, based on their presence in food and their
 affinity for the aryl hydrocarbon receptor (AhR): benzo(a)pyrene (BP),
 dibenzo(a,h)anthracene (DBA), and pyrene (PYR). Treatment of primary rat
 and WIF-B9 hepatocytes by all three PAHs increased the release of EVs,
 mainly comprised of exosomes, in parallel with modifying exosome protein
 marker expression and inducing apoptosis. Moreover, PAH treatment of
 rodents for three months also led to increased EV levels in plasma. The EV
 release involved CYP metabolism and the activation of the transcription
 factor, the AhR, for BP and DBA and another transcription factor, the
 constitutive androstane receptor (CAR), for PYR. Furthermore, all PAHs
 increased cholesterol levels in EVs but only BP and DBA were able to
 reduce the cholesterol content of total cell membranes. All cholesterol
 changes very likely participated in the increase in EV release and cell
 death. Finally, we studied changes in cell membrane fluidity caused by BP
 and DBA due to cholesterol depletion. Our data showed increased cell
 membrane fluidity, which contributed to hepatocyte EV release and cell
 death.
description: Figure
 S1Figure S1: Dose and time
 response exposure in terms of cell death and proliferation in WIF-B9 cells
 upon PAH exposure in terms of cell death and
 proliferation.Figure
 S2Figure S2. Original
 pictures of western-blots from which some results are extracted for a
 presentation in figure 2C.Figure
 S3Figure S3 : PAHs do not
 affect the size of EVs released by WIF-B9 cells.Figure
 S4Figure S4 : PAHs do not
 affect the size of EVs released by primary rat
 hepatocytes.Figure
 S5Figure S5 : mRNA
 expression of CYP1A1, CYP1A2, CYP1B1 and CYP2C6 in WIF-B9 cells and
 primary rat hepatocytes.Figure
 S6Figure S6 : CYP1
 activity in WIF-B9 and primary rat hepatocytes.Figure
 S7Figure S7 : Cell death
 and EV release of WIF-B9 cells and primary rat hepatocytes exposed to AhR
 agonist TCDD.Figure
 S8Figure S8 : Effects of
 pravastatin on cholesterol content of hepatocytes (A,B) and EVs (E) and
 also on bulk membrane fluidity (C,D).Supplementary Table
 S1Table S1: Selected PAH
 properties.Supplementary Table
 S2Table S2: Concentration
 of PAHs in human serum and daily intake of PAHs.Supplementary Table
 S3Table S3: Pre-treatment
 of WIF-B9 and primary rat hepatocytes.Supplementary Table
 S4Table S4 : Culture
 medium volume needed for EV isolation.Supplementary Table
 S5Table S5 : List of
 primers used for RT-qPCR experiments.