Adipose tissue–derived stromal cells’ conditioned medium modulates endothelial‐mesenchymal transition induced by IL‐1β/TGF‐β2 but does not restore endothelial function

Abstract

Objectives Endothelial cells undergo TGF‐β–driven endothelial‐mesenchymal transition (EndMT), representing up to 25% of cardiac myofibroblasts in ischaemic hearts. Previous research showed that conditioned medium of adipose tissue–derived stromal cells (ASC‐CMed) blocks the activation of fibroblasts into fibrotic myofibroblasts. We tested the hypothesis that ASC‐CMed abrogates EndMT and prevents the formation of adverse myofibroblasts. Materials and methods Human umbilical vein endothelial cells (HUVEC) were treated with IL‐1β and TGF‐β2 to induce EndMT, and the influence of ASC‐CMed was assessed. As controls, non‐treated HUVEC or HUVEC treated only with IL‐1β in the absence or presence of ASC‐CMed were used. Gene expression of inflammatory, endothelial, mesenchymal and extracellular matrix markers, transcription factors and cell receptors was analysed by RT‐qPCR. The protein expression of endothelial and mesenchymal markers was evaluated by immunofluorescence microscopy and immunoblotting. Endothelial cell function was measured by sprouting assay. Results IL‐1β/TGF‐β2 treatment induced EndMT, as evidenced by the change in HUVEC morphology and an increase in mesenchymal markers. ASC‐CMed blocked the EndMT‐related fibrotic processes, as observed by reduced expression of mesenchymal markers TAGLN (P = 0.0008) and CNN1 (P = 0.0573), as well as SM22α (P = 0.0501). The angiogenesis potential was impaired in HUVEC undergoing EndMT and could not be restored by ASC‐CMed. Conclusions We demonstrated that ASC‐CMed reduces IL‐1β/TGF‐β2‐induced EndMT as observed by the loss of mesenchymal markers. The present study supports the anti‐fibrotic effects of ASC‐CMed through the modulation of the EndMT process.

SOX9 acts as a dynamic pioneer factor inducing stable changes in the chromatin landscape to reprogram endothelial cells

“…1C). This change in morphology is consistent with other studies’ observations of EndMT in HUVECs (Yu et al 2017; Liguori et al 2019; Noseda et al 2004). Furthermore, we observed enhanced migratory properties of the HUVECs after SOX9 expression, as shown by transwell assays (Fig.…”

Nur77 deficiency exacerbates cardiac fibrosis after myocardial infarction by promoting endothelial‐to‐mesenchymal transition

“…In our study, 1 month after MI, the mRNA expression of TGFβ2 was still upregulated compared with WT control, implying TGFβ2's important role in cardiac remodeling in the long term (Figure 2e). It has been demonstrated that IL-1β and TGFβ2 acted synergistically to induced EndMT in human umbilical vein endothelial cells (Liguori, Liguori, Moreira, & Harmsen, 2019;Maleszewska et al, 2013) and this effect is also confirmed in HCAECs in our study. Taken together, these results indicated that the inflammatory microenvironment after MI potentiated EndMT.…”

Calcium-Sensing Receptor Participates in High Glucose-Induced EndMT in Primary Human Aortic Endothelial Cells

“…HAECs are commonly recognized as a substitute for the basic research study of endothelial cells. Fourth, numerous researches had proved that HG-induced EndMT contributed to endothelial damage ( Maleszewska et al, 2013 ; Liguori et al, 2019 ; Guan et al, 2020 ), which lead to cardiovascular diseases in diabetes. Thus, our study focus on the role of CaSR in HG-induced EndMT, which might underlie the mechanism cardiovascular diseases in diabetes.…”

“…High levels of serum glucose is believed to be the key to promoting the association between endothelial damage and cardiovascular disease ( Giugliano et al, 2019 ; Joffre et al, 2020 ; Liberale et al, 2020 ). Previous studies have shown that high glucose (HG) induces endothelial cell (EC) damage via endothelial-to-mesenchymal transition (EndMT) ( Maleszewska et al, 2013 ; Liguori et al, 2019 ), which contributes to endothelial injury and subsequent cardiovascular disease in DM. However, the underlying mechanisms of HG-induced EndMT remain largely unclear.…”