title: MicroRNA-30b is a multifunctional regulator of aortic valve interstitial cells.
abstract: OBJECTIVE
  Calcific aortic valve disease is an active process involving a wide range of pathologic changes. Valve interstitial cells are the most prevalent cells in the heart valve and maintain normal valve structure and function. MicroRNAs (miRNAs) are essential posttranscriptional modulators of gene expression, and miRNA-30b is a known repressor of bone morphogenetic protein 2-mediated osteogenesis. We hypothesized that miRNA-30b is a multifunctional regulator of aortic valve interstitial cells during calcification.
  
  
  METHODS
  To determine the role of miRNA-30b in calcific aortic valve disease, we evaluated miRNA expression in human calcific aortic valve leaflets obtained intraoperatively. Furthermore, human valve interstitial cells were evaluated with regard to miRNA-30b expression and osteogenesis by quantitative real-time polymerase chain reaction, Western blotting, flow cytometry, and alkaline phosphatase assays.
  
  
  RESULTS
  In this study, we demonstrated that miRNA-30b attenuates bone morphogenetic protein 2-induced osteoblast differentiation by targeting Runx2, Smad1, and caspase-3. Transfection of a mimic of miRNA-30b led to decreases in alkaline phosphatase activity and expressions of Runx2, Smad1, and caspase-3. Furthermore, dual luciferase reporter assays confirmed that Runx2, Smad1, and caspase-3 are direct targets of miRNA-30b.
  
  
  CONCLUSIONS
  We demonstrated a remarkable role of miRNA-30b in calcific aortic valve disease as a regulator of human aortic valvular calcification and apoptosis through direct targeting of Runx2, Smad1, and caspase-3. Targeting of miRNA-30b could serve as a novel therapeutic strategy to limit progressive calcification in aortic stenosis.
authors: Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors: Mi
authors: M
authors: Zhang
authors: Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors: Xiaohong
authors: X
authors: Liu
authors: Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors: Xiwu
authors: X
authors: Zhang
authors: Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors: Zhigang
authors: Z
authors: Song
authors: Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors: Lin
authors: L
authors: Han
authors: Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
authors: Yuanyuan
authors: Y
authors: He
authors: Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China. Electronic address: zhiyunx@hotmail.com.
authors: Zhiyun
authors: Z
authors: Xu
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D001021:Aortic Valve / Q000378:metabolism* / Q000473:pathology*; D001024:Aortic Valve Stenosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D002114:Calcinosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D010012:Osteogenesis; D015398:Signal Transduction; D051898:Smad1 Protein / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D015415:Biomarkers; D050659:Core Binding Factor Alpha 1 Subunit; C546086:MIRN30b microRNA, human; D035683:MicroRNAs; C493709:RUNX2 protein, human; C497399:SMAD1 protein, human; D051898:Smad1 Protein; D000469:Alkaline Phosphatase; C505407:CASP3 protein, human; D053148:Caspase 3