Impact of the flame retardant 2,2'4,4'-tetrabromodiphenyl ether (PBDE-47) in THP-1 macrophage-like cell function via small extracellular vesicles

2,2'4,4'-tetrabromodiphenyl ether (PBDE-47) is one of the most widespread environmental brominated flame-retardant congeners which has also been detected in animal and human tissues. Several studies have reported the effects of PBDEs on different health issues, including neurobehavioral and developmental disorders, reproductive health, and alterations of thyroid function. Much less is known about its immunotoxicity. The aim of our study was to investigate the effects that treatment of THP-1 macrophage-like cells with PBDE-47 could have on the content of small extracellular vesicles' (sEVs) microRNA (miRNA) cargo and their downstream effects on bystander macrophages. To achieve this, we purified sEVs from PBDE-47 treated M(LPS) THP-1 macrophage-like cells (sEVs(PBDE+LPS)) by means of ultra-centrifugation and characterized their miRNA cargo by microarray analysis detecting the modulation of 18 miRNAs. Furthermore, resting THP-1 derived M(0) macrophage-like cells were cultured with sEVs(PBDE+LPS), showing that the treatment reshaped the miRNA profiles of 12 intracellular miRNAs. This dataset was studied in silico, identifying the biological pathways affected by these target genes. This analysis identified 12 pathways all involved in the maturation and polarization of macrophages. Therefore, to evaluate whether sEVs(PBDE+LPS) can have some immunomodulatory activity, naive M(0) THP-1 macrophage-like cells cultured with purified sEVs(PBDE+LPS) were studied for IL-6, TNF-alpha and TGF-beta mRNAs expression and immune stained with the HLA-DR, CD80, CCR7, CD38 and CD209 antigens and analyzed by flow cytometry. This analysis showed that the PBDE-47 treatment does not induce the expression of specific M1 and M2 cytokine markers of differentiation and may have impaired the ability to make immunological synapses and present antigens, down-regulating the expression of HLA-DR and CD209 antigens. Overall, our study supports the model that perturbation of miRNA cargo by PBDE-47 treatment contributes to the rewiring of cellular regulatory pathways capable of inducing perturbation of differentiation markers on naive resting M(0) THP-1 macrophage-like cells.

Automated summary

This study investigates the effects of PBDE-47 treatment on the miRNA cargo of sEVs and their downstream effects on bystander macrophages. The sEVs purified from PBDE-47 treated cells were found to modulate the expression of 12 miRNAs in resting THP-1 cells. Furthermore, PBDE-47 treatment may impair immune activity and down-regulate differentiation markers in macrophages. These findings suggest that PBDE-47 perturbs cellular regulatory pathways and affects macrophage differentiation markers by altering miRNA cargo.