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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
MicroRNA-17-5p, a novel endothelial cell modulator, controls vascular re-endothelialization and neointimal lesion
Xiaopei Liu1, Jing Chen1, Gen Liu1
1Department of Cardiology, Cardiovascular Research Institute, Hubei Key Laboratory of Cardiology, 117921Renmin Hospital of Wuhan University, Wuhan, China.
Background:
The functions of miR-17-5p in tumorigenesis have been explored. However, their functionalities in arterial endothelial cells (ECs) have not been investigated. Besides, the issue of vascular remodelling is barely addressed.
Objectives:
The study aimed to determine the effect of overexpression or inhibition of miR-17-5p on arterial endothelial cells' (ECs) function and vascular remodelling in vitro and the rat carotid arteries model.
Methods:
Quantitative RT-PCR analysis was performed to examine the expression of miR-17-5p. Then, gain-of-function and loss-of-function approaches were employed to investigate the functional roles of miR-17-5p in cultured human coronary artery endothelial cells (HCAECs); further, TargetScan software analysis and luciferase reporter activity assay were performed to investigate the potential mechanism. Lastly, the results of the cell segment were verified in a rat carotid artery balloon injury model by Western blot analysis, measurement of the vascular cGMP level and plasma 8-iso-prostaglandin F2 (8-iso-PGF2) testing. Moreover, morphometric analysis was implemented to detect the re-endothelialization and neointimal formation in rat carotid artery after balloon injury.
Results:
This study firstly found that miR-17-5p expression was upregulated in the injured vascular walls and highly expressive in ECs; overexpression of miR-17-5p inhibited HCAECs' proliferation and migration, whereas miR-17-5p knockdown strengthened its proliferative and migratory roles, influenced inflammatory response, through regulating VEGRA and VEGFR2. It was found that miR-17-5p bind to VEGFA and VEGFR2 at the 3'UTR. Next, downregulation of miR-17-5p promotes re-endothelialization, and attenuates neointimal formation as measured by the I/M ratio (0.63±0.05 vs 1.45±0.06, antagomiR-17-5p vs. Lenti-NC, p < 0.05). In addition, the functional recovery of the endothelium was also accelerated by miR-17-5p knockdown.
Conclusion:
Our study suggests that miR-17-5p is a feasible strategy for the selective modulation of endothelialization and vascular remodelling through regulating VEGFA and VEGFR2.
Insights
MicroRNA-17-5p (miR-17-5p) regulates vascular remodeling. Inhibiting miR-17-5p promotes re-endothelialization and reduces neointimal formation, offering a therapeutic strategy for vascular repair.
Area of Science:
- Vascular Biology
- Molecular Biology
- Biochemistry
Background:
- MicroRNA-17-5p (miR-17-5p) functions in tumorigenesis are known, but its role in arterial endothelial cells (ECs) and vascular remodeling remains unexplored.
- Vascular remodeling is a critical process in cardiovascular health and disease, yet specific molecular regulators are not fully understood.
Purpose of the Study:
- To investigate the impact of miR-17-5p modulation on arterial endothelial cell function and vascular remodeling.
- To elucidate the underlying molecular mechanisms involving VEGFA and VEGFR2.
Main Methods:
- Quantitative RT-PCR to assess miR-17-5p expression.
- Gain- and loss-of-function studies in human coronary artery endothelial cells (HCAECs).
- In vivo validation in a rat carotid artery balloon injury model, including morphometric analysis and molecular assays.
Main Results:
- miR-17-5p expression is upregulated in injured vascular walls and ECs.
- Overexpression of miR-17-5p inhibits EC proliferation and migration; knockdown enhances these functions by regulating VEGFA and VEGFR2.
- Downregulation of miR-17-5p promotes re-endothelialization and attenuates neointimal formation in vivo.
Conclusions:
- miR-17-5p plays a significant role in regulating endothelialization and vascular remodeling.
- Targeting miR-17-5p offers a promising therapeutic strategy for modulating vascular repair processes.
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