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Updated: Aug 9, 2025
Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In Vitro
Published on: May 10, 2021
Inhibition of miR-101-3p prevents human aortic valve interstitial cell calcification through regulation of CDH11/SOX9
Jianglei Chen1, Yi Lin1, Zhongjie Sun2,3
1Department of Physiology, College of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA.
Insights
MicroRNA-101-3p promotes calcific aortic valve disease (CAVD) by upregulating osteogenesis pathways in human aortic valve interstitial cells (HAVICs). Inhibiting miR-101-3p may offer a therapeutic strategy for CAVD.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Biochemistry
Background:
- Calcific aortic valve disease (CAVD) is a significant cause of adult heart disease.
- Understanding the molecular mechanisms of CAVD is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role of miR-101-3p in the calcification of human aortic valve interstitial cells (HAVICs).
- To elucidate the underlying molecular mechanisms by which miR-101-3p influences HAVIC calcification.
Main Methods:
- Small RNA deep sequencing and qPCR were employed to analyze microRNA expression in calcified human aortic valves.
- Primary HAVICs were cultured and treated with miR-101-3p mimics and inhibitors to assess effects on calcification and osteogenesis.
- Luciferase reporter assays and expression analysis were used to identify and validate miR-101-3p targets.
Main Results:
- miR-101-3p expression was significantly increased in calcified human aortic valves.
- Overexpression of miR-101-3p promoted HAVIC calcification and osteogenesis, while its inhibition reduced these processes.
- miR-101-3p directly targets and downregulates cadherin-11 (CDH11) and SOX9, key regulators of osteogenesis.
Conclusions:
- miR-101-3p plays a critical role in HAVIC calcification by regulating CDH11 and SOX9 expression.
- miR-101-3p represents a potential therapeutic target for the treatment of calcific aortic valve disease.
Background:
Calcific aortic valve disease (CAVD) is the second leading cause of adult heart diseases. The purpose of this study is to investigate whether miR-101-3p plays a role in the human aortic valve interstitial cells (HAVICs) calcification and the underlying mechanisms.
Methods:
Small RNA deep sequencing and qPCR analysis were used to determine changes in microRNA expression in calcified human aortic valves.
Results:
The data showed that miR-101-3p levels were increased in the calcified human aortic valves. Using cultured primary HAVICs, we demonstrated that the miR-101-3p mimic promoted calcification and upregulated the osteogenesis pathway, while anti-miR-101-3p inhibited osteogenic differentiation and prevented calcification in HAVICs treated with the osteogenic conditioned medium. Mechanistically, miR-101-3p directly targeted cadherin-11 (CDH11) and Sry-related high-mobility-group box 9 (SOX9), key factors in the regulation of chondrogenesis and osteogenesis. Both CDH11 and SOX9 expressions were downregulated in the calcified human HAVICs. Inhibition of miR-101-3p restored expression of CDH11, SOX9 and ASPN and prevented osteogenesis in HAVICs under the calcific condition.
Conclusion:
miR-101-3p plays an important role in HAVIC calcification through regulation of CDH11/SOX9 expression. The finding is important as it reveals that miR-1013p may be a potential therapeutic target for calcific aortic valve disease.
