YY1 promotes diabetic vascular calcification via m⁶A-Binding protein HNRNPC

Diabetes is one of the most common and fastest-growing diseases worldwide, and diabetic atherosclerotic calcification is a frequent and fatal complication, the underlying mechanisms of which remain unclear. In this study, we investigated the mechanism by which HNRNPC regulates diabetic vascular calcification.

We retrieved datasets GSE211722, GSE84012, and GSE74755 from the GEO database and performed probe-to-gene name conversion. Based on expression profile data and aortic transcriptomic data from diabetic and non-diabetic mice, RNA m6A methylation-related regulatory genes were screened, and violin plots and heatmaps were generated accordingly. Differential genes identified from the expression profiles and experimental groups were subjected to intersection analysis to ultimately identify the target gene. We also enrolled coronary heart disease patients meeting predefined inclusion criteria to analyze the correlation between coronary artery calcium scores and serum HNRNPC levels. Subsequently, through transcription factor prediction and validation using single-cell transcriptomic data from the anterior tibial arteries of diabetic amputation patients, transcription factors of HNRNPC were identified. The role of HNRNPC in diabetic atherosclerotic calcification was further investigated by establishing an in vitro model of smooth muscle cells under high-glucose conditions and an in vivo model of diabetic atherosclerotic calcification in ApoE^-/- mice.

Analysis of GEO datasets and diabetic mouse transcriptomic data identified HNRNPC as the only overlapping differentially expressed m6A methylation regulatory gene from both sources. Clinical investigations revealed that serum HNRNPC levels and coronary artery calcium scores were elevated in diabetic patients and exhibited a positive correlation. In vitro and in vivo experiments demonstrated that inhibiting HNRNPC reduced the expression of the osteogenic marker RUNX2 and decreased calcium deposition, whereas HNRNPC overexpression promoted calcification. By integrating bioinformatics analysis with cellular (MOVAS) and animal (ApoE^-/- mice) models, the transcription factor YY1 was revealed to play a pivotal role in vascular calcification. During calcification, YY1 expression was upregulated, and it directly bound to and activated the HNRNPC promoter, thereby enhancing HNRNPC transcription. Silencing YY1 significantly suppressed calcium deposition and osteogenic marker expression, whereas overexpressing HNRNPC reversed this effect, confirming that YY1 drives smooth muscle cell calcification by regulating HNRNPC expression.

We demonstrate a previously unrecognized role of HNRNPC as a key driver of diabetic atherosclerotic calcification. Specifically, the transcription factor YY1 promotes calcification by mediating HNRNPC's function.