BACKGROUND: Calcific aortic valve disease is characterized by an abnormal 
mineralization of the aortic valve. Osteogenic activity in the aortic valve is 
under the control of NOTCH1, which regulates the expression of key 
pro-osteogenic genes such as RUNX2 and BMP2. Long noncoding RNAs (lncRNAs) may 
reprogram cells by altering the gene expression pattern.
METHODS: Multidimensional genomic profiling was performed in human aortic valves 
to document the expression of lncRNAs and the DNA methylation pattern in 
calcific aortic valve disease. In-depth functional assays were carried out to 
document the impact of lncRNA on the mineralization of the aortic valve.
RESULTS: We documented that lncRNA H19 (H19) was increased in calcific aortic 
valve disease. Hypomethylation of the promoter region was observed in 
mineralized aortic valves and was inversely associated with H19 expression. 
Knockdown and overexpression experiments showed that H19 induces a strong 
osteogenic phenotype by altering the NOTCH1 pathway. Gene promoter analyses 
showed that H19 silenced NOTCH1 by preventing the recruitment of p53 to its 
promoter. A knockdown of H19 in valve interstitial cells (VICs) increased the 
expression of NOTCH1 and decreased the level of RUNX2 and BMP2, 2 downstream 
targets repressed by NOTCH1. In rescue experiments, the transfection of a vector 
encoding for the active Notch intracellular domain prevented H19-induced 
mineralization of valve interstitial cells.
CONCLUSIONS: These findings indicate that a dysregulation of DNA methylation in 
the promoter of H19 during calcific aortic valve disease is associated with a 
higher expression of this lncRNA, which promotes an osteogenic program by 
interfering with the expression of NOTCH1.