MiR-1249 on Endothelial Extracellular Vesicles Mediates Cigarette Smoke–Induced Pulmonary Hypertension by Inhibiting HDAC10 (Histone Deacetylase 10)-NFκB (Nuclear Factor κB)-CaSR (Calcium-Sensing Receptor) Cascade

Abstract

Background: Overproduction of endothelial extracellular vesicles (eEVs) is correlated with pulmonary hypertension progression, but the precise mechanism remains largely unclear. Methods: MicroRNA-chip and real-time polymerase chain reaction real-time polymerase chain reaction were conducted to screen and validate microRNA profiles in blood plasma eEVs of rats and human with or without cigarette smoking. Pulmonary artery smooth muscle cells were cultured to study signaling pathways. Pulmonary hypertension phenotypes were evaluated in wild-type and calcium-sensing receptor knockout rats to identify the pathophysiological significance of the microRNA pathway. Results: MicroR-1249 was predominant highly expressed in eEVs from plasma of rats exposed to cigarette smoking, and confirmed in eEVs from plasma of human smokers as well as in eEVs from cigarette smoke extract-treated pulmonary artery endothelial cells, but not in cigarette smoke extract-treated pulmonary artery smooth muscle cells. In cultured pulmonary artery smooth muscle cells, microR-1249 downregulated the expression of histone deacetylase 10, which in turn enhanced the acetylated form of NFκB (nuclear factor κB) level and its nuclear translocation leading to increased expression of calcium-sensing receptor. In rats, the repression of microR-1249 in eEVs by microR-1249 inhibitor, histone deacetylase 10 overexpression, or calcium-sensing receptor knockout profoundly inhibited the proliferative capacities and diminished apoptosis-resistance of pulmonary artery smooth muscle cells and pulmonary hypertension development in rats intravenously administrated with eEVs preparation from cigarette smoke extract-treated pulmonary artery endothelial cells. Conclusions: Cigarette smoke–enriched microR-1249 in endothelial extracellular vesicles facilitates the hyperproliferative and antiapoptotic status of pulmonary artery smooth muscle cells promoting pulmonary hypertension evolution through the inhibition of histone deacetylase 10–NFκB–calcium-sensing receptor cascade.

The HDAC10 instructs macrophage M2 program via deacetylation of STAT3 and promotes allergic airway inflammation

“…First, the Hdac10 fl/fl -LysMCre mice were conditionally knocked out for HDAC10 in macrophages (CKO). HDAC10 is expressed in various types of cells, such as epithelial cells, endothelial cells and T cells 29 , 30 . Further experiments are needed to investigate the effect of other cells on HDAC10 in the regulation of BMDMs and the role of HDAC10 in other cells.…”

Emerging Epigenetic Targets and Their Molecular Impact on Vascular Remodeling in Pulmonary Hypertension

“…Importantly, this process is mediated by epigenetic histone acetylation [30]. Other studies determined that endothelial-secreted factors promote the hyperproliferation of SMCs using epigenetic mechanisms such as histone acetylation and microRNA [18,31]. Future exploration is needed to further determine the epigenetic contribution that cell-cell interaction has to PH vascular remodeling and to assess the reversibility of epigenetic targeting in PH (Figure 1).…”

Extracellular Vesicles in Pulmonary Hypertension: A Dangerous Liaison?

“…Focusing on EVs as targets of treatment, growing evidence highlights the possible benefits of blocking the release of specific extravesicular miRNAs. When exposed to cigarette smoke, endothelial cells release EVs enriched in miR-1249 which promotes PASMCs hyperproliferation and favors antiapoptotic status [ 56 ]. Chronic hypoxia, utilized to induce PH in rats as well in a variety of PH animal models [ 71 ], not only provokes hemodynamic and pathologic changes but also increased the number of circulating exosomes and the levels of exosomal miR-211 [ 57 ].…”