Figure1.

DIANA clinical study pipeline. DIANA clinical study (NCT04371809) aimed to investigate specific DNA methylation changes in circulating CD4⁺ and CD8⁺ T cells in patients with ACS as compared to HS. Genomic DNA, extracted from the two cellular subsets, was analysed through RRBS. Then, significant DMRs were identified and annotated. Functional pathway analysis was used to identify the most prevalent pathways subtending ACS. Gene expression validation of the DMGs most prevalent in disease pathways was performed by qRT-PCR. Molecular data were correlated with clinical and biochemical parameters. A biological network was built with the aim to visualize protein–protein, co-expression and functional interactions.

ACS: acute coronary syndromes; HS: healthy subjects; RRBS: reduced-representation bisulphite sequencing; DMG: differentially methylated gene; DMR: differentially methylated region; q-RT-PCR: quantitative real-time polymerase chain reaction; PPI: protein–protein interaction.