European Atherosclerosis Journal

Postprandial implications in cardiovascular disease and potential markers to develop new therapies

2025-07-30T17:08:52+00:00

An unbalanced diet significantly raises the risk of various chronic diseases and cancers, contributing to increased morbidity and mortality globally. Today, the link between metabolic status and cardiovascular disease is well established. Disruptions in glucose and lipid homeostasis, particularly postprandial hyperglycemia and hyperlipidemia are key risk factors for cardiovascular conditions. These postprandial metabolic disturbances promote atherosclerosis and cardiovascular injury, primarily by triggering endothelial dysfunction.
Lifestyle interventions play a pivotal role, and pharmacological treatments aimed at controlling lipid and glucose levels generally lead to improvements in both fasting and postprandial states. However, further research is necessary to establish reference values for biomarkers of postprandial dysmetabolism and to evaluate their clinical relevance. Individuals who exhibit a mismatch between fasting and postprandial levels of glucose and triglycerides, namely, those with normal or mildly elevated fasting levels but exaggerated postprandial responses, may represent a subgroup at heightened and potentially modifiable risk for both microvascular and macrovascular complications. Validating biomarkers of postprandial dysmetabolism could offer valuable clinical tools for improved risk assessment and personalized therapeutic strategies. This review summarises the unique physiology of triglyceride-rich lipoprotein metabolism after meals and the disruptions that can foster cardiovascular complications. Given the scarcity of targeted therapies, it also discusses emerging treatment candidates and their underlying mechanisms.

Comparing the predictive value of genetic determinants and measured plasma levels of Lipoprotein(a) in cardiovascular risk assessment: evidence from a large-scale UK Biobank study

2025-09-01T07:06:55+00:00

Introduction: Lipoprotein(a) [Lp(a)] is a genetically influenced lipoprotein causally associated with atherosclerotic cardiovascular disease risk. This study compares the predictive value of genetically determined versus directly measured Lp(a) levels for major coronary events (MCE).
Methods: From UK Biobank data, participants with complete genetic and plasma Lp(a) data, including LPA variants rs3798220 and rs10455872, were selected. Cox proportional hazards models were employed to estimate the risk of MCE, associated with both Lp(a) genetic score (0, 1, or ≥2 minor alleles) and measured Lp(a) levels.
Results: Among 410,194 participants (mean age 57.25, 54% females), both Lp(a) genetic score and measured levels were independently associated with a stepwise increase in MCE risk. Within each genetic score group, increasing measured Lp(a) quintiles were associated with higher MCE. However, for individuals with similar measured Lp(a), MCE risk did not differ by genetic score.
Conclusions: Directly measured Lp(a) levels offer superior cardiovascular risk prediction, supporting the practice of measuring Lp(a) levels at least once in adulthood.

Evaluation of the Effect of Lomitapide Treatment on Major Adverse Cardiovascular Events (MACE) in Patients with Homozygous Familial Hypercholesterolemia: Study Protocol of the LILITH Study

2025-09-01T06:59:31+00:00

Homozygous Familial Hypercholesterolemia (HoFH) is a rare genetic disorder characterized by markedly elevated low-density lipoprotein cholesterol (LDL-C) levels from birth, leading to premature and severe cardiovascular disease. Lomitapide, an inhibitor of microsomal triglyceride transfer protein (MTP), effectively lowers LDL-C in HoFH patients. However, data on its impact on major adverse cardiovascular events (MACE) remain limited, and randomized controlled trials are not feasible due to the rarity of the condition and ethical constraints. This article presents the protocol of the LILITH study (Evaluation of the Effect of Lomitapide Treatment on Major Adverse Cardiovascular Events in Patients with Homozygous Familial Hypercholesterolemia), a multicenter, observational, retrospective–prospective cohort study. The study aims to compare the incidence of MACE during the first three years of lomitapide treatment with that observed in the three years preceding treatment, within the same cohort of adult HoFH patients (target N=72). Clinical data, including MACE, lipid levels, liver function, safety outcomes, and concomitant lipid-lowering therapies, will be collected. The primary analysis will apply McNemar’s test to assess changes in MACE incidence pre- and post-treatment. This methodological approach enables the evaluation of long-term cardiovascular outcomes in a real-world setting for a rare disease population.

TechNote - Minimum Preanalytical Information for the Publication of Studies on Circulating Cell-Free microRNA-based Biomarkers

2025-07-30T17:02:11+00:00

Circulating cell-free microRNAs (miRNAs) are emerging as promising biomarkers with broad potential for clinical applications. However, pre-analytical variability significantly affects miRNA quantification and hampers reproducibility across studies. This work aims to define the Minimum Preanalytical Information required for the publication of studies on circulating cell-free miRNA-based biomarkers. We review key pre-analytical factors that influence circulating miRNA levels quantified using RT-qPCR. Critical variables include blood collection timing, sample type, centrifugation protocols, transport and storage conditions, hemolysis, lipemia, medication, physical activity and pathogen inactivation methods. We introduce a standardized checklist to promote methodological transparency and inter-study comparability. The final aim is to enhance the reliability of miRNA-based biomarker research and support its successful translation into clinical practice.

Progress toward Implementing Multiomic Approaches in Atherosclerotic Cardiovascular Disease: Update from the 4th AtheroNET Meeting in Sarajevo (Bosnia and Herzegovina)

2025-09-01T12:58:22+00:00

The COST Action CA21153 “Network for Implementing Multiomic Approaches in Atherosclerotic Cardiovascular Disease Prevention and Research” (AtheroNET; https://atheronet.eu) was launched in October 2022 with the mission to accelerate the application of multiomics in atherosclerosis research and to foster collaboration among experts from diverse disciplines. Born from the pressing need to advance atherosclerosis management and overcome existing challenges, AtheroNET creates a collaborative arena where innovation thrives through dialogue. The initiative was spearheaded by Prof. Miron Sopić (Faculty of Pharmacy, University of Belgrade), the primary proposer of the Action. The Action is led by Chair Prof. Paolo Magni (Università degli Studi di Milano) and Vice-Chair Prof. Yvan Devaux (Luxembourg Institute of Health), with strategic support from a dedicated leadership team:
• Grant Holder Scientific Representative: Dr. Ines Potočnjak
• Science Communication Coordinator: Prof. Georgios Kararigas
• Grant Awarding Coordinator: Dr. Susana Novella
• WG1 - Pathophysiological mechanisms: applying multiomics to uncover novel pathogenic players and processes; Leader: Prof. Dimitris Kardassis
• WG2 - Personalized clinical models: translating omics insights into improved management of ASCVD; Leader: Prof. Alberico Catapano
• WG3 - Standardization and harmonization: developing SOPs and guidelines to enhance reproducibility across omics research; Leader: Dr. Marie Mardal
• WG4 - Data integration and ML/AI: optimizing algorithms for integrating complex multiomic datasets; Leader: Dr. Aleksandra Gruca
• WG5 - Dissemination and communication: sharing advances with scientists, clinicians, patients, and the public; Leader: Prof. Georgios Kararigas
Since its inception, AtheroNET has grown to 475 members from across Europe, embracing COST’s core values of inclusiveness and excellence across geography, gender, and career stage. A significant proportion of members come from Inclusive Target Countries (n=282), with strong representation of women (n=277) and early-career researchers (n=223).

Functional and metabolomic characterization of human cell models to address obesity and metabolic dysfunction- associated steatotic liver disease pathogenetic mechanisms

2025-09-02T18:48:36+00:00

Introduction and aim. Obesity and metabolic dysfunction-associated steatotic liver disease (MASLD) are global health concerns linked to an increased risk of atherosclerotic cardiovascular disease (ASCVD). Despite progresses in elucidating relevant pathophysiological mechanisms involved in obesity and MASLD, a comprehensive understanding toward ASCVD risk is missing due to the lack of reliable in vitro models reproducing obesity-related dysfunctional adipose tissue and MASLD.
This study focuses on developing and characterizing innovative in vitro models recapitulating key features of these conditions assessing novel molecular signatures for potential drug targeting.
Methods. Human-derived SW872 adipocytes, dysfunctional SW872 (SW872-OA) obtained by 7- days 100 μM oleic acid (OA), 17-days spontaneously differentiated SW872 (SW872-AUTO), human hepatoma HepG2 cells, and a HepG2-based steatotic-like model induced with 7-days 100 μM OA (HepG2-OA), were selected. All in vitro models were characterized using ¹H NMR spectroscopy, light-microscopy, spectrophotometry, flow cytometry, and RT-qPCR.
Results and Discussion. Metabolomic and lipidomic analysis allowed to identify specific metabolite signatures associated with dysfunctional patterns in both SW872 and HepG2 cell models. SW872- OA, SW872-AUTO, and HepG2-OA models showed increased lipid/triglycerides accumulation, confirmed by light-microscopy observations, and spectrophotometry quantification. HepG2-OA cells exhibited elevated triglyceride synthesis genes expression. Functional assays revealed reduced glucose uptake and elevated ROS production across all dysfunctional models. Trolox antioxidant treatment mitigated ROS levels in SW872 and HepG2 cells with minor effects on SW872-OA, SW872-AUTO, and HepG2-OA. Gene expression analysis (GEA) showed significant upregulation of oxidative stress-related genes in HepG2-OA cells. GEA in SW872-OA and SW872-AUTO cells revealed upregulation of genes involved in adipocyte differentiation, inflammation, and glycemic homeostasis.
Conclusion. These results showed that OA-treatment and SW872-spontaneous differentiation generate in vitro models recapitulating dysfunctional adipose tissue and MASLD.
Relevance for AtheroNET. Dysfunctional SW872 and HepG2 cell models show specific molecular, metabolomic and lipidomic signatures which may translate to innovative pharmacological targets for obesity, MASLD, and ASCVD.

Effects of Pro-Resolving Lipid Mediators on Nitric Oxide and Prostacyclin Pathways in Human Endothelial Cells

2025-09-03T10:39:07+00:00

Introduction and Aim: Inflammation plays a central role in the development and progression of atherosclerosis. Pro- resolving lipid mediators, including Resolvins (RvE1) and Maresins (MaR1) are key regulators of the resolution phase of inflammation. Despite their known anti-inflammatory effects, their impact on nitric oxide (NO) and prostacyclin (PGI2) levels in human umbilical vein endothelial cells (HUVECs) remains underexplored. This study investigates the effects of RvE1 and MaR1 on vascular inflammation, focusing on NO and PGI2 in HUVECs.
Methods: An in vitro inflammatory model was established by incubating HUVECs with lipopolysaccharides (LPS, 100 µg/mL) and interleukin-1 beta (IL-1β, 100 ng/mL) for 24 hours. RvE1 and MaR1 (100 nM) were then applied under the same conditions. Levels of tumor necrosis factor-alpha (TNF-α), total nitrite-nitrate, and the stable PGI2 metabolite 6- keto-PGF1α were measured in the incubation medium using ELISA.
Results and Discussion: LPS and IL-1β significantly increased TNF-α levels, confirming the inflammatory response. RvE1 and MaR1 significantly reduced nitrite-nitrate levels, suggesting their role in mitigating vascular inflammation. However, no significant changes were observed in 6-keto-PGF1α levels (n=3-4, p>0.05). These findings highlight the selective effects of RvE1 and MaR1 on different pathways involved in endothelial inflammation.
Conclusion: This preliminary study demonstrates that RvE1 and MaR1 appear to reduce nitrite-nitrate levels, their lack of impact on 6-keto-PGF1α suggests distinct pathway-specific actions that warrant further investigation. Future studies with larger sample sizes, varied inflammatory conditions, are essential to confirm these findings and better understand their therapeutic potential.
Relevance to AtheroNET: This research aligns with AtheroNET's goal of exploring novel anti-inflammatory strategies, emphasizing the importance of pro-resolving lipid mediators in vascular health.

This work was supported by funding from Scientific and Technological Research Council of Türkiye (TÜBİTAK) (Project number: 1919B012313469)

Proteomic analysis for prediction of type 2 diabetes identifies cardiovascular disease-related proteins

2025-09-03T10:07:50+00:00

Introduction and aim: Cardiovascular disease (CVD) and type 2 diabetes (T2D) are interconnected chronic conditions which cause significant global health challenges and mortality. T2D is a risk factor for CVD characterized by insulin resistance and beta-cell dysfunction. This study aims to investigate the association between the blood plasma proteome and T2D in the Trøndelag Health (HUNT) Study.
Methods: The HUNT Study is a population-based cohort with four waves of enrollment beginning in 1984. 5,402 samples from HUNT3 were analysed with SomaScan including 3,221 on SomaScanv4.0 (~5,000 proteins) and 2,181 on SomaScanv4.1 (~7,000 proteins). T2D was defined based on self-reported disease from questionnaires. Binary logistic regression analysis was performed to test the associations of protein concentrations with T2D. Models included adjustment by sex, age, waist-hip-ratio (WHR), smoking status, and comorbidities.
Results and Discussion: We identified 584 proteins that are significantly associated with T2D. From these proteins, CILP2 and MXRA8 are associated with decreased risk, while PLXB2 and NFASC are associated with increased risk for having T2D. The most significant pathway from GO enrichment analysis includes proteins related to lipid catabolic process (FDR adjusted p- value: 0.000054). Indeed, several proteins significantly associated with T2D are known for their role in lipid metabolism, for instance Adiponectin (OR 0.55; 95% CI 0.46 - 0.65), Apo A – IV (1.57;1.37 - 1.81), Apo B (0.63; 0.55 - 0.73), Apo C – I (0.73; 0.64 - 0.84), and Apo D (0.49;0.38 - 0.63).
Conclusion: The proteins and pathways identified represent insights into the underlying molecular mechanism of T2D and may serve as potential biomarkers for risk prediction and prevention for CVD.

Relevance for AtheroNET: The findings suggest that early identification of these proteins could mitigate cardiovascular risks in individuals with or at risk for T2D.

A validated and reproducible LC-MS/MS analytical method to quantify an emerging cardiovascular risk biomarker, trimethylamine N-Oxide (TMAO), in human plasma

2025-09-02T18:03:14+00:00

Introduction and aim. The interplay between nutrition patterns, gut microbiota-derived metabolites and cardiovascular diseases (CVDs) has recently become a field of intensive research. Gut microbiota-derived metabolites represent an attractive source of biomarkers for CVDs, including atherosclerotic cardiovascular disease (ASCVD) and metabolic diseases. Trimethylamine (TMA), a gut-derived metabolite, undergoes oxidation in the liver, resulting in trimethylamine N-oxide (TMAO). Recent evidence from in vivo studies and clinical trials has correlated TMAO with increased CVD risk and occurrence due to the atherogenic potential of TMAO. TMAO has been quantified by exploiting mass spectrometry (MS) techniques, coupled either with liquid (LC) or gas chromatography (GC), and nuclear magnetic resonance (NMR) spectroscopy. However, standardization parameters and conditions still represent a critical issue when dealing with data reproducibility and robustness.
Hence, this study aimed at developing a validated and highly reproducible method for the quantification of TMAO in human-derived plasma.
Methods. A high-performance LC (HPLC) system (Shimadzu) coupled to tandem MS (MS/MS) (Sciex QTRAP 6500+) was selected, along with multiple reaction monitoring (MRM) modality and electrospray ionization (ESI) in positive polarity. Calibration curve was obtained spiking a healthy subject serum sample with different TMAO concentrations in the ng/mL range.

Results and Discussion. The HPLC-MS/MS method was optimized according to the different MS parameters and by selecting the most appropriate column for HPLC assessment. Method validation was performed evaluating the intra-/inter-assay accuracy and precision. The coefficient of variation (CV%) always resulted below 20%.
Conclusion. The HPLC-MS/MS method showed to be robust and reproducible according to standardization requirements, and useful to assess ASCVD risk in a cohort with subclinical atherosclerosis, food intake record and microbiome evaluation¹.
Relevance for AtheroNET. TMAO data from characterized CVD patient cohorts, obtained in a reproducible and standardized manner, could be integrated by artificial intelligence/machine learning-based approach to improve CVD risk stratification toward precision medicine.