The asialoglycoprotein receptor 1, the glycome, and proteome associate with coronary atherosclerosis progression on serial photon-counting coronary computed tomography angiography

Introduction Current biomarkers for coronary artery disease (CAD) fail to adequately predict disease progression or recurrent cardiovascular events. The asialoglycoprotein receptor 1 (ASGR1), which clears circulating glycans (sugars), is emerging as a novel regulator of atherosclerosis. The relationship between ASGR1, the glycome and CAD progression remains unexplored. Aim To investigate associations between ASGR1, the N-glycome, and related proteome with coronary atherosclerosis progression in a human cohort with symptomatic CAD. Methods One-hundred CAD patients with residual atheroma were prospectively recruited for simultaneous serial blood biobanking and coronary plaque imaging with serial photon-counting coronary computed tomography angiography (PC-CCTA) and quantitative plaque burden analysis, at baseline and 12-months follow-up. Isolated peripheral blood mononuclear cells (PBMCs) were assessed for: (1) ASGR1 protein by ELISA, (2) their N-glycome and (3) proteome by mass spectrometry. Healthy Control participants (N= 31) were free from atheroma. Plaque progression was defined as a ≥2% in total plaque burden. Results In this high-risk cohort, with median age 62 years, 76% male, 85% recent acute coronary syndrome, 95% on statins at follow-up, serial PC-CCTA revealed dynamic responsiveness of residual coronary atheroma one-year after invasive angiography (Figure 1). 39% of patients were plaque Progressors, 32% had Stable disease and 29% were Regressors. Clinical risk factors and baseline or achieved low-density lipoprotein cholesterol levels did not differ between Regressors or Progressors. However, higher ASGR1 protein levels demonstrated a positive trend with CAD progression (median in Healthy 0.096 ng/mL, Regressors 0.091 ng/mL, Stable 0.098 ng/mL, Progressors 0.138 ng/mL; P-trend = 0.031) (Figure 2). A doubly-sialylated N-glycan (S2F1.a) was overexpressed in CAD versus Healthy Control PBMCs, which remained elevated in Progressors (Figure 3). Two oligomannose N-glycans were also lower in the Progressors compared to Healthy Controls (Figure 3). Accompanying proteomics revealed 363 differentially regulated proteins between Healthy Controls versus CAD (Figure 4) with metabolic and inflammatory patterns with mechanistic links to CAD pathobiology (Figure 5). Amongst the individual significant proteins in CAD patients, a protein phosphatase-regulator and autophagy-promoter, Immunoglobulin Binding Protein 1, was downregulated relative to Healthy Controls (Figure 6). In another comparison Thymosin beta-4 was upregulated in Regressors, relative to Progressors, at baseline. Conclusions Although clinical factors did not differentiate coronary atheroma progression status on quantitative PC-CCTA, differences were identified in PBMC ASGR1, their N-glycome and proteome. Our multi-omics findings implicate ASGR1 and the glycome as new biomarkers of CAD progression, and highlights known proteins as ready therapeutic targets.Picture 1Picture 2