Multi-omics of in vitro aortic valve calcification

Area of Science:

• Cardiovascular Biology
• Molecular Biology
• Biochemistry

Background:

• Heart valve calcification, specifically calcific aortic valve disease (CAVD), is a complex process involving the osteogenic differentiation of valve interstitial cells (VIC).
• The precise molecular mechanisms driving VIC osteogenic differentiation in CAVD remain incompletely understood, necessitating further investigation.

Purpose of the Study:

• To compare the molecular mechanisms of VIC osteogenic differentiation in healthy individuals versus patients with CAVD.
• To identify key genes, pathways, and molecular players involved in the early and later stages of VIC osteogenic differentiation.

Main Methods:

• Utilized RNA-sequencing (RNA-seq) transcriptomics at 48 hours and shotgun proteomics at 10 days to analyze human VIC.
• Employed bioinformatic analysis to identify differentially expressed genes and proteins and their associated signaling pathways.
• Validated the role of ZBTB16 (also known as PLZF) through lentiviral overexpression and subsequent calcification assays.

Main Results:

• Identified numerous stage-specific differentially expressed genes and proteins with good correlation between transcriptomic and proteomic data.
• Revealed involvement of signaling pathways including PI3K-Akt, MAPK, Ras, and TNF in VIC osteogenic differentiation.
• Highlighted the early involvement of Wnt, FoxO, and HIF-1 signaling pathways in VIC commitment and observed metabolic pathway shifts.
• Demonstrated that ZBTB16 overexpression significantly increased VIC calcification, suggesting a pro-osteogenic role.

Conclusions:

• VIC osteogenic differentiation in CAVD is regulated by a complex interplay of signaling and metabolic pathways.
• ZBTB16 emerges as a critical pro-osteogenic factor in the early stages of VIC differentiation and calcification.
• These findings provide novel insights into the molecular pathogenesis of CAVD and potential therapeutic targets.