KEYNOTE: Mechanisms of Calcific Aortic Valve Disease Visualized Through the Lens of Multi-Omics

Calcific aortic valve disease (CAVD) is a progressive and multifactorial condition marked by fibrosis, inflammation, and osteogenic remodeling of the aortic valve, culminating in stenosis and heart failure. Traditional paradigms have often relied on reductionist approaches, limiting insight into the spatial and cellular complexity of the disease. In contrast, proteomics offers a holistic lens to dissect the molecular underpinnings of CAVD at tissue, cellular, and subcellular levels. This presentation highlights recent advances in multi-omic-scale analysis of human aortic valves, including spatially resolved profiling across disease stages (non-diseased, fibrotic, and calcific regions) and anatomical layers (fibrosa, spongiosa, and ventricularis). These studies reveal layer-specific and stage-specific enrichment of proteins involved in fibrosis, inflammation, and mineralization. Integration with single-cell RNA seq further uncovers a heterogeneous population of valve interstitial cells (VICs), including a progenitor-like, disease-driving population marked by high CD44 and CD29 expression. This subpopulation exhibits distinct calcification trajectories upon osteogenic stimulation, supported by time-course proteomics and pseudotime analysis. Moreover, extracellular vesicles (EVs) have emerged as critical mediators of disease progression. Proteomic analysis of tissue-resident EVs identifies cargoes linked to pro-calcific signaling. The application of multi-omics, machine learning, and network biology has enabled the construction of a Proteome Atlas of CAVD, providing a systems-level framework to identify and validate novel therapeutic targets, including MAOA, CTHRC1, SORT1 and MFSD5. Together, these studies demonstrate that proteomics, in concert with transcriptomics, single-cell RNA seq and functional assays, enables unprecedented resolution in visualizing CAVD pathogenesis. This integrative strategy paves the way for precision medicine approaches to treat a disease historically considered irreversible.