Plasma Small Extracellular Vesicle-Carried miRNA-501-5p Promotes Vascular Smooth Muscle Cell Phenotypic
Xiao-Fei Gao1,2, Zhi-Mei Wang1, Ai-Qun Chen1
1Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
Abstract:
Vascular smooth muscle cell (VSMC) phenotypic modulation plays an important role in the occurrence and development of in-stent restenosis (ISR), the underlying mechanism of which remains a key issue needing to be urgently addressed. This study is designed to investigate the role of plasma small extracellular vesicles (sEV) in VSMC phenotypic modulation. sEV were isolated from the plasma of patients with ISR (ISR-sEV) or not (Ctl-sEV) 1 year after coronary stent implantation using differential ultracentrifugation. Plasma sEV in ISR patients are elevated markedly and decrease the expression of VSMC contractile markers α-SMA and calponin and increase VSMC proliferation. miRNA sequencing and qRT-PCR validation identified that miRNA-501-5p was the highest expressed miRNA in the plasma ISR-sEV compared with Ctl-sEV. Then, we found that sEV-carried miRNA-501-5p level was significantly higher in ISR patients, and the level of plasma sEV-carried miRNA-501-5p linearly correlated with the degree of restenosis (R 2 = 0.62). Moreover, miRNA-501-5p inhibition significantly increased the expression of VSMC contractile markers α-SMA and calponin and suppressed VSMC proliferation and migration; in vivo inhibition of miRNA-501-5p could also blunt carotid artery balloon injury induced VSMC phenotypic modulation in rats. Mechanically, miRNA-501-5p promoted plasma sEV-induced VSMC proliferation by targeting Smad3. Notably, endothelial cells might be the major origins of miRNA-501-5p. Collectively, these findings showed that plasma sEV-carried miRNA-501-5p promotes VSMC phenotypic modulation-mediated ISR through targeting Smad3.
Insights
Plasma extracellular vesicles carrying miRNA-501-5p promote in-stent restenosis by altering vascular smooth muscle cell (VSMC) behavior. Inhibiting this miRNA could be a therapeutic strategy for preventing restenosis after stenting.
Area of Science:
- Cardiovascular Biology
- Extracellular Vesicles
- Molecular Medicine
Background:
- Vascular smooth muscle cell (VSMC) phenotypic modulation is critical in in-stent restenosis (ISR).
- The precise mechanisms driving VSMC modulation in ISR, particularly involving plasma-derived factors, require further elucidation.
- Small extracellular vesicles (sEV) are increasingly recognized as key mediators of intercellular communication.
Purpose of the Study:
- To investigate the role of plasma small extracellular vesicles (sEV) in vascular smooth muscle cell (VSMC) phenotypic modulation related to in-stent restenosis (ISR).
- To identify specific molecular cargo within sEVs that contributes to ISR pathogenesis.
- To explore the therapeutic potential of targeting these sEV-mediated pathways.
Main Methods:
- Isolation of sEV from plasma of ISR patients and controls using differential ultracentrifugation.
- miRNA sequencing and qRT-PCR to identify differentially expressed miRNAs in ISR-sEV.
- In vitro studies assessing the impact of miRNA-501-5p on VSMC proliferation, migration, and marker expression.
- In vivo studies using a rat carotid artery balloon injury model to evaluate miRNA-501-5p inhibition.
- Mechanistic studies to determine the downstream target of miRNA-501-5p.
Main Results:
- Plasma sEV levels were elevated in ISR patients and induced VSMC proliferation while decreasing contractile markers.
- miRNA-501-5p was the most highly expressed miRNA in ISR-sEV and its levels correlated with restenosis severity.
- Inhibition of miRNA-501-5p reversed VSMC phenotypic modulation in vitro and in vivo.
- miRNA-501-5p promotes VSMC proliferation by targeting Smad3, with endothelial cells identified as potential origin.
- Plasma sEV-derived miRNA-501-5p drives VSMC modulation-mediated ISR.
Conclusions:
- Plasma sEV-derived miRNA-501-5p is a key driver of VSMC phenotypic modulation contributing to ISR.
- Targeting miRNA-501-5p within plasma sEVs represents a promising therapeutic strategy for preventing in-stent restenosis.
- Endothelial cells may be a significant source of pro-restenotic miRNA-501-5p in the plasma.
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