CD40 levels in plasma are associated with cardiovascular disease and in carotid plaques with a vulnerable plaque phenotype and remodelling

Abstract

Background and purpose CD40 and CD40 ligand (CD40L) are costimulatory molecules and members of the TNF receptor superfamily well known for their involvement in inflammatory and autoimmune diseases. This study uses two large human cohorts – the SUrrogate markers for Micro- and Macro-vascular hard endpoints for Innovative diabetes Tools (SUMMIT) and the Carotid Plaque Imaging Project (CPIP) – to explore the potential of plasma or intra-plaque expression of CD40 and CD40L as biomarkers and to locally affect plaque stability. Methods Proximity Extension Assay (PEA) technique was used to measure soluble CD40 and CD40L (sCD40 and sCD40L) in plasma from 1437 subjects from the SUMMIT cohort, the majority of which (80%) with pre-existing cardiovascular disease, and in atherosclerotic plaque homogenates from 199 subjects of the CPIP cohort undergoing carotid endarterectomy. The Mann-Whitney U test was used to compare groups and Spearman's rank correlation/the Chi-square test was used to assess correlations. Multiple comparisons were corrected for using the Holm-Šídák test. A logistic regression model was used to test for associations with future cardiovascular events and mortality. Results In the SUMMIT cohort both plasma CD40 and CD40L levels were elevated in individuals with a history of stroke (p=0.000030 and p=0.020, respectively), while sCD40 levels also were higher in individuals with a prior acute myocardial infarction (p=0.016). Plasma levels of sCD40 correlated with carotid plaque burden (as measured by ultrasound imaging, r=0.355, p<1x10–16) and were associated with future cardiovascular events over a three year-follow up period (p=0.02, hazard ratio 1.3, 95% C.I: 1.042–1.625). sCD40 and sCD40L were associated with a plaque phenotype characterized by the strong presence of features both of vulnerability such as high content oxidized low-density lipoprotein (LDL; r=0.236, p=0.004 and r=0.259, p=0.0037, respectively) and pro-inflammatory cytokines (e.g. tumour necrosis factor-α: p=3.1x10–7 and p=0.0006, respectively) and low calcium content (r=−0.208, p=0.012 and r=0.268, p=0.00034, respectively). Conclusion High plasma CD40 and CD40L levels are associated with symptomatic cardiovascular disease. Plasma CD40 levels correlate with the severity of carotid atherosclerosis and are associated with an increased risk for future cardiovascular events. Additionally, intra-plaque levels are associated with a vulnerable plaque phenotype. Our findings thus support the value of sCD40 and sCD40L both as biomarkers and therapeutic targets for cardiovascular disease. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): The Swedish Heart and Lung Foundation (1) and the Swedish Research Council (2)

Soluble CD40 receptor is a biomarker of the burden of carotid artery atherosclerosis in subjects at high cardiovascular risk

“…Human data in support of the role of soluble CD40R as a marker of systemic atherosclerosis are becoming consistent. We have already reported that the expression of CD40R is high in carotid atherosclerosis plaques and correlates with the plaque lipid content, particularly with oxidized LDL (51) . More recently, through an extensive human plaque RNA expression analysis, CD40R was found to be one of the 4 most expressed proteins (52) .…”

“…Interestingly, by adding to conventional risk factors LDL-cholesterol the area under the curve of ROC analysis for presence of any plaque showed no significant change (from 0.7914 to 0.7910). Although the data need to be replicated in other cohorts, recent data from the SUMMIT 3-year follow-up indicate that CD40R, as a continuous variable, is an independent predictor of cardiovascular events and its concentration in plaque homogenates -collected from a different cohort -correlates with histologic indices of plaque vulnerability (51) . We acknowledge that the quantification of atherosclerosis was limited to carotid arteries, thus we cannot conclude that CD40R can actually predict the atherosclerosis burden of other territories, however atherosclerosis is frequently a systemic disease and, in fact, the presence of plaques in the carotid artery is considered also by the most recent guidelines (61) a risk modifier for cardiac events and recent studies found close associations between serum CD40R and both coronary artery (52,54) and peripheral atherosclerosis (53) .…”

Roles of Macrophages in Atherogenesis

“…Plasma CD40 levels are associated with the severity of carotid AS and an increased risk for future CV events. Intraplaque levels of sCD40 and sCD40L are also associated with vulnerability and remodeling (Shami et al, 2020). The costimulatory CD40 and CD40L are implicated in the regulation of the inflammatory response during AS development.…”

Sensing Soluble Immune Checkpoint Molecules and Disease-Relevant Cytokines in Cancer: A Novel Paradigm in Disease Diagnosis and Monitoring

“…This highly stable and accurate immunosensor exhibited a DL of 13.1 pg/mL for sCD40L (Yuan et al, 2015). It is important to note here, that sCD40L is an important biomarker in cardio-vascular diseases as well (Shami et al, 2020) and therefore, biosensor-mediated detection of soluble isoforms of immune-checkpoint molecules has broader applications in many diseases.…”