We aimed to investigate whether the expression of the OPG/RANK/RANKL triad in peripheral blood mononuclear cells (PBMC) and circulating levels of markers of ectopic mineralization (OPG, FGF-23, PPi) are modified in patients with calcific aortic valve disease (CAVD). We found that patients affected by CAVD (n = 50) had significantly higher circulating levels of OPG as compared to control individuals (p = 0.003). No differences between the two groups were found in FGF-23 and PPi levels. RANKL expression was higher in the PBMC from CAVD patients (p = 0.018) and was directly correlated with the amount of valve calcification (p = 0.032). In vitro studies showed that treatment of valve interstitial cells (VIC) with RANKL plus phosphate was followed by increase in matrix mineralization (p = 0.001). In conclusion, RANKL expression is increased in PBMC of patients with CAVD, is directly correlated with the degree of valve calcification, and promotes pro-calcific differentiation of VIC.
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Conflict of Interest
The authors declare that they have no conflict of interest.
Informed Consent Statement
All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000. Informed consent was obtained from all subjects for being included in the study.
Associate Editor Adrian Chester oversaw the review of this article.
Electronic Supplementary Material
Correlation between Pi and RANKL levels with the calcification of aortic valve. The gene expression of RANKL in PBMC and the levels of Pi in the serum have been correlated with the degree of calcium deposition within the aortic valve (n = 38). In particular, patients with CAVD were stratified into four groups according to both RANKL and Pi levels. Patients were considered RANKL_high or RANKL_low in case of ΔCT values below or above the median, respectively. Similarly patients were classified as Pi_high or Pi_low in case of serum Pi levels above or below the median of the population. The percentage of calcified valve areas was quantified in CT images by using a customized image analysis system (see also Fig. 2). Although without reaching statistical significance (ANOVA p = 0.074), we observed that the higher degree of calcified area was observed in patients with both RANKL_high and Pi_high. Viceversa, the lower percentage of calcification was seen in the group of patients with both RANKL_low and Pi_low. (PNG 47 kb)
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Rattazzi, M., Faggin, E., Bertacco, E. et al. RANKL Expression Is Increased in Circulating Mononuclear Cells of Patients with Calcific Aortic Stenosis. J. of Cardiovasc. Trans. Res. 11, 329–338 (2018). https://doi.org/10.1007/s12265-018-9804-2