Cardiovascular Disease Biomarkers Research Articles

The Potential Contribution of MyomiRs miR-133a-3p, -133b, and -206 Dysregulation in Cardiovascular Disease Risk

Cardiovascular disease (CVD) is a major global health concern. The number of people with CVD is expected to rise due to aging populations and increasing risk factors such as obesity and diabetes. Identifying new molecular markers is crucial for early diagnosis and treatment. Among these, plasma levels of some miRNAs, specifically expressed in cardiac and skeletal muscle, known as myomiRs, have gained attention for their roles in cardiovascular health. This study analyzed the plasma levels of miR-133a-3p, -133b, and -206 in the pathogenesis of cardiovascular diseases. Using a case–control study design with patients recruited from several nursing homes from Calabria (southern Italy) characterized by different types of CVD compared with non-CVD controls, we found downregulation of miR-133a-3p in heart failure and miR-133b in stroke, along with the overall decreased expression of miR-133b and miR-206 in CVD patients, although they showed low specificity as biomarkers of CVD (as based on ROC analysis). In silico functional characterization of their targets and signaling pathways revealed their involvement in critical cardiovascular processes. Although further research is necessary to fully elucidate their mechanisms and clinical utility, the findings reported here may provide insight into the potential contribution of myomiRs in the cardiovascular injury framework, also offering indications for new research directions.

Multifaceted Role of Apolipoprotein C3 in Cardiovascular Disease Risk and Metabolic Disorder in Diabetes

Apolipoprotein C3 (APOC3) plays a critical role in regulating triglyceride levels and serves as a key predictor of cardiovascular disease (CVD) risk, particularly in patients with diabetes. While APOC3 is known to inhibit lipoprotein lipase, recent findings reveal its broader influence across lipoprotein metabolism, where it modulates the structure and function of various lipoproteins. Therefore, this review examines the complex metabolic cycle of APOC3, emphasizing the impact of APOC3-containing lipoproteins on human metabolism, particularly in patients with diabetes. Notably, APOC3 affects triglyceride-rich lipoproteins and causes structural changes in high-, very low-, intermediate-, and low-density lipoproteins, thereby increasing CVD risk. Evidence suggests that elevated APOC3 levels—above the proposed safe range of 10–15 mg/dL—correlate with clinically significant CVD outcomes. Recognizing APOC3 as a promising biomarker for CVD, this review underscores the urgent need for high-throughput, clinically feasible methods to further investigate its role in lipoprotein physiology in both animal models and human studies. Additionally, we analyze the relationship between APOC3-related genes and lipoproteins, reinforcing the value of large-population studies to understand the impact of APOC3 on metabolic diseases. Ultimately, this review supports the development of therapeutic strategies targeting APOC3 reduction as a preventive approach for diabetes-related CVD.

Evaluation of diagnostic performances of Pro-neurotensin and Heart-type fatty acid binding protein as reliable biomarkers for cardiovascular diseases

ABSTRACT Aim In 2019, cardiovascular diseases accounted for 32% of global deaths. So, early detection of cardiac disorders is crucial. The study aimed to examine the suitability of Pro-neurotensin and Heart-type fatty acid binding protein as dependable biomarkers for cardiac patients with Heart failure as a primary diagnosis. Methodology The prospective study involved 204 Egyptian volunteers (100 cardiac patients and 104 controls) enrolled from El-Sahel Teaching Hospital in Cairo, Egypt, between October 2022 and May 2023. Inclusion criteria included a high risk of cardiovascular events with symptoms like a fast or irregular pulse, shortness of breath, and fatigue. Exclusion criteria included asymptomatic individuals, cognitive disorders, and psychiatric conditions. The Research Ethics Committee approved the protocol. The consultant conducted a clinical examination of all patients and assessed their heart state. Serum ProNT and H-FABP were detected using a kit for the sandwich ELISA technique. Results ProNT and H-FABP were significantly elevated in patients compared to controls with p < 0.001. Demonstrated sensitivity of 81% and 84%, with a specificity of 89% and 91%, respectively. Conclusion Elevated ProNT and H-FABP levels are associated with severe CVDs, suggesting their potential as diagnostic biomarkers for patients, specifically those with heart failure, as a primary characteristic.

The quality, uniqueness, and causality of NMR-based prediction models for low-density lipoprotein cholesterol subfractions in human blood plasma

Low-density lipoprotein (LDL) cholesterol (chol) subfractions are risk biomarkers for cardiovascular diseases (CVD). A reference analysis, ultracentrifugation (UC), is laborious and may be replaced with a rapid prediction using proton NMR spectra of human blood plasma. However, the quality and uniqueness of these prediction models of biologically related subfractions remains unknown. This study, using two independent cohorts (n = 277), investigates the inter-correlations between LDL cholesterol in the main fraction and five subfractions, as well as the independence of their NMR-based prediction models. The results reveal that the prediction models utilize both shared and unique spectral information from the NMR spectra to determine concentrations of LDL subfractions. Analysis of variance contributions for prediction and causality assessments demonstrate that the NMR spectra contain unique predictive information for the LDL1chol, LDL2chol, and LDL5chol subfractions. In contrast, the spectral signatures for LDL3chol and LDL4chol are either insufficient or confounded. Our findings indicate that these five CVD biomarkers represent two independent clusters, reflecting their biosynthetic pathways, and confirm the presence of causal relationships between certain LDL chol subfractions. This highlights the importance of employing caution when interpreting the concentrations of specific LDL subfractions as standalone biomarkers for CVD risk.

Colorimetric aptasensors for sensitive low-density lipoprotein detection based on reduced oxide graphene@molybdenum disulfide-ferrocene nanosheets with peroxidase-like activity.

Low-density lipoprotein (LDL) is a key biomarker for cardiovascular disease (CVD) risk assessment. Monitoring LDL for the early diagnosis of CVD and its complications is an important clinical analysis tool. In this work, a novel colorimetric aptasensor for LDL detection was constructed via reduced graphene oxide@molybdenum disulfide-ferrocene-carboxylic nanosheets (rGO@MoS2-Fc) with excellent peroxidase-like activity. On this basis, the LDL aptamer (LDLapt) immobilized on the surface of rGO@MoS2-Fc served as a signal probe (rGO@MoS2-Fc/LDLapt), while the unmodified LDLapt served as a capture probe. When LDL was present, it was recognized by the LDLapt and rGO@MoS2-Fc/LDLapt to form an rGO@MoS2-Fc/LDLapt/LDL/LDLapt sandwich-type conjugate with excellent enzymatic catalytic properties that can catalyze the generation of hydroxyl radicals (·OH) from hydrogen peroxide (H2O2), which in turn oxidized the colorless substrate o-phenylenediamine (OPD) to the yellow compound 2,3-diamino phenothiazine (DAP). In addition, the catalytic mechanism of the reaction was confirmed to be induced by ·OH through free radical experiments. The aptasensor had a linear range of 15.0 to 200.0 μg mL-1, and a limit of detection (LOD) of 2.199 μg mL-1. Overall, the assay has high selectivity, sensitivity and operability, showing broad application prospects in the clinical diagnosis of CVD.

Insulin-like Growth Factor-Binding Protein-1 (IGFBP-1) as a Biomarker of Cardiovascular Disease.

Insulin-like growth factor-binding protein-1 (IGFBP-1) contributes to the regulation of IGFs for metabolism and growth and has IGF-independent actions. IGFBP-1 in the circulation is derived from the liver, where it is inhibited by insulin and stimulated by multiple factors, including proinflammatory cytokines. IGFBP-1 levels are influenced by sex and age, which also determine cardiometabolic risk and patterns of disease presentation. While lower circulating IGFBP-1 concentrations are associated with an unfavorable cardiometabolic risk profile, higher IGFBP-1 predicts worse cardiovascular disease outcomes. This review explores these associations and the possible roles of IGFBP-1 in the pathophysiology of atherosclerosis. We recommend the evaluation of dynamic approaches, such as simultaneous measurements of fasting IGFBP-1 and proinsulin level in response to an oral glucose challenge, as well as multi-marker approaches incorporating markers of inflammation.

Nonobese young females with PCOS are at high risk for long-term cardiovascular disease.

Whether polycystic ovary syndrome (PCOS) is an independent risk factor for long-term cardiovascular disease (CVD) is unclear, and the risk of CVD in easily overlooked young nonobese PCOS patients is unknown. This study aimed to investigate the associations of PCOS with CVD and identify the management priorities. 3864 participants (645 with PCOS) from UK Biobank were recruited from 2006-2010. The cumulative incidences of the CVD were calculated and compared between patients with and without PCOS via the log rank test. Cox proportional risk regression models were used to assess the relationships of PCOS with CVD and the impact of PCOS treatments on CVD risk. Polygenic risk scores and linkage disequilibrium score regression were used to assess the genetic-level associations. Then proteomics subgroup cohort was conducted to explore the significant biomarker involved in the PCOS-CVD associations. Compared with participants without PCOS, participants with PCOS had greater risks of CVD (hazard ratio (HR)=1.77, 95% confidence interval (CI)=1.19-2.65), coronary artery disease (HR=2.27, 95% CI=1.35-3.81) and myocardial infarction (HR=2.08, 95% CI=1.11-3.90) independent of genetic risk, especially for young nonobese PCOS patients (Pfor interaction <0.05). Current commonly used treatments did not affect CVD incidence. Proteomics cohort revealed that discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2) may be specific CVD biomarker for patients with PCOS. Patients with PCOS had an increased risk of CVD, and young nonobese PCOS patients should be prioritized for CVD risk management. These findings support the necessity of clinical surveillance and suggest DCBLD2 as a possible CVD biomarker in females with PCOS.

Ceramides as Biomarkers of Cardiovascular Diseases and Heart Failure.

Ceramides are lipid species that play several physiological roles in the body, including stress response, inflammation, and apoptosis, and their involvement in lipid metabolism and energy production makes them crucial in the pathophysiology of heart failure (HF). Several species of ceramides and ceramide signatures have recently been investigated as possible biomarkers of cardiovascular disease (CVD), and risk scores have demonstrated prognostic value in stratifying patients by risk and possibly predicting adverse cardiac events. With growing interest in targeting metabolic dysfunction, understanding the role of ceramides in CVD also opens the possibility of novel therapeutics that target ceramide metabolism to improve cardiac function and cardiac outcomes in patients. Understanding the role of ceramides in CVD opens the possibility of novel diagnostics and theragnostic targeting ceramide metabolism to improve cardiac function and cardiac outcomes in patients with heart failure.

Gut Microbial Taxonomy and Its Role as a Biomarker in Aortic Diseases: A Systematic Review and Future Perspectives.

Background/Objectives: Evidence of the association between the gut microbiome and cardiovascular diseases has accumulated. An imbalance or dysbiosis of this system has been shown to play a role in the pathogenesis of cardiovascular events, including aortic diseases. We aimed to elucidate the findings of the gut microbial taxonomy associated with aortic diseases and their subtypes. Furthermore, we sought to investigate whether gut microbiome dysbiosis can be used as a biomarker for aortic disease detection and to identify which species can be disease-specific. Methods: A systematic search was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines for original research papers on gut microbiome composition in patients with aortic disease, using patients without aortic disease as the control (i.e., healthy controls). The databases PubMed, Scopus, Cochrane, and Web of Science were used by employing the medical subject headings (MeSH) terms "aortic diseases", "microbiome"," microbiota", and "taxa" before August 2024. We extracted the study characteristics, study population, and gut microbiome in aortic disease, including microbiota taxa diversity and abundance, regardless of taxa level. The National Institutes of Health (NIH) Quality Assessment Tool was used to assess study quality. Data were synthesized narratively to address the heterogeneity of the studies. Results: In this review, twelve studies that have identified gut microbial species and their potential impact on aortic disease pathogenesis were included. The studies showed the phyla dominance of Bacillota, Pseudomonadota, Actinomycetota, Bacteroidota, and Euryarchaeota in aortic disease patients. We also included the taxa sequencing methods and those used to extract the microorganisms. Aortic diseases were categorized into Takayasu's arteritis, giant cell arteritis, aortic aneurysm, and aortic dissection. Aortic disease patients had a higher rate of dysbiosis when compared to the healthy control groups, with significantly different microbiome composition. Conclusions: Patients with aortic disease exhibit a distinct difference between their gut microbiota composition and that of the healthy controls, which suggests a potential biomarker role of gut dysbiosis. Further exploration of the microbiome and its metagenome interface can help identify its role in aortic disease pathogenesis in depth, generating future therapeutic options. However, a unified methodology is required to identify potential microbial biomarkers in cardiovascular and cardiometabolic diseases.

Innovative biomarkers in cardiovascular disease: Advances in prevention and therapeutic

Innovative biomarkers in cardiovascular disease: Advances in prevention and therapeutic

Exploring the roles of SPARC as a proinflammatory factor and its potential as a novel therapeutic target against cardiovascular disease.

Cardiovascular disease (CVD) is a leading cause of death worldwide, and the number of patients with CVD continues to increase despite extensive research and developments in this field. Chronic inflammation is a pivotal pathological component of CVD, and unveiling new proinflammatory factors will help devise novel preventive and therapeutic strategies. The extracellular matrix (ECM) not only provides structural support between cells but also contributes to cellular functions. Secreted protein acidic and rich in cysteine (SPARC) is a collagen-binding matricellular protein that is particularly induced during development and tissue remodeling. A proinflammatory role for SPARC has been demonstrated in various animal models, such as in the lipopolysaccharide-induced footpad model and dextran sodium sulfate-induced colitis model. Recent clinical studies reported a positive correlation between elevated plasma SPARC levels and hypertension, obesity, and the inflammatory marker high-sensitive C-reactive protein. In addition, SPARC gene deletion attenuates the cardiac injury induced by aging, myocardial infarction, and pressure load, suggesting that SPARC has deleterious effects on CVD. This review summarizes the regulatory and proinflammatory mechanisms of SPARC on CVD, chronic kidney disease (CKD), and cerebrovascular disease and discusses the rationale behind measuring SPARC as a biomarker of CVD and the effects of inhibition of SPARC in the prevention and treatment of CVD.

Serum megalin levels in type-2 diabetes mellitus with and without cardiovascular disease

ABSTRACT Background: Low-density lipoprotein receptor-related protein-2 (LRP2), also called megalin, is a multi-ligand receptor of the LDL receptor gene family mediating reabsorption of ligands like Apo-A1. Type 2 diabetes mellitus (T2DM) may possibly disrupt megalin functions as it is found to be regulated by insulin. This might cause cardiovascular complications due to derangement in lipoprotein metabolism. The current study was carried out to assess the serum megalin levels among T2DM individuals with cardiovascular complications in the Indian population. Methods: This was a cross-sectional study involving 80 patients with T2DM. 40 T2DM patients with known cardiovascular disease were selected as cases and 40 of those without evidence of cardiovascular disease were selected as controls. Demographic details, DM duration and class of oral hypoglycemic agents (OHA) used were collected from medical records while details of lipid profile, fasting glucose, serum creatinine and urea were collected from laboratory information system. Serum megalin levels were estimated using left-over samples by ELISA. Results: The study groups showed no statistical significance in baseline laboratory parameters except for serum creatinine and low density lipoprotein cholesterol (LDL-c). Mean serum megalin levels were statistically insignificant between cases and controls (0.91 ± 0.78 ng/mL vs. 0.85 ± 0.69 ng/mL, P 0.74).The subgroup analysis of serum megalin levels based on OHA consumption among cases was also statistically insignificant (P 0.056). Pearson’s correlational analysis was statistically insignificant between serum megalin and lipid profile parameters among cases (P &gt; 0.05). Conclusion: Serum megalin alone may not serve as a biomarker for cardiovascular disease.

Fibroblast Growth Factors in Cardiovascular Disease.

Despite advancements in managing traditional cardiovascular risk factors, many cardiovascular diseases (CVDs) persist. Fibroblast growth factors (FGFs) have emerged as potential diagnostic markers and therapeutic targets for CVDs. FGF1, FGF2, and FGF4 are primarily used for therapeutic angiogenesis. Clinical applications are being explored based on animal studies using approaches such as recombinant protein administration and adenovirus-mediated gene delivery, targeting patients with coronary artery disease and lower extremity arterial disease. Although promising results have been observed in animal models and early-stage clinical trials, further studies are required to assess their therapeutic potential. The FGF19 subfamily, consisting of FGF19, FGF21, and FGF23, act via endocrine signaling in various organs. FGF19, primarily expressed in the small intestine, plays important roles in glucose, lipid, and bile acid metabolism and has therapeutic potential for metabolic disorders. FGF21, found in various tissues, improves glucose metabolism and insulin sensitivity, suggesting potential for treating obesity and diabetes. FGF23, primarily secreted by osteocytes, regulates vitamin D and phosphate metabolism and serves as an important biomarker for chronic kidney disease and CVDs. Thus, FGFs holds promise for both therapeutic and diagnostic applications in metabolic and cardiovascular diseases. Understanding the mechanisms of FGF may pave the way for novel strategies to prevent and manage CVDs, potentially addressing the limitations of current treatments. This review explores the roles of FGF1, FGF2, FGF4, and the FGF19 subfamily in maintaining cardiovascular health. Further research and clinical trials are crucial to fully understand the therapeutic potential of FGFs in managing cardiovascular health.

Serum neurofilament light chain: a novel biomarker for cardiovascular diseases in individuals without hypertension

Serum neurofilament light chain (sNFL) is a biomarker for axonal injury. Previous studies have linked sNFL levels to cardiovascular risk factors such as diabetes and hypertension, but its association with cardiovascular diseases (CVD) remains unclear. This study aims to explore the association between sNFL and CVD and evaluates its predictive value. Utilizing NHANES 2013–2014 data, this study included 2,035 participants aged ≥ 20 years with measured sNFL quantified using a Siemens immunoassay. CVD was self-reported and included myocardial infarction, stroke, heart failure, coronary heart disease, or angina. Logistic regression models assessed the association between sNFL levels and CVD. The predictive value of sNFL for CVD was evaluated using area under the curve (AUC) and DeLong test. Participants with higher sNFL levels were typically older, male, non-Hispanic white, smokers, and had lower socioeconomic status, higher CVD, hypertension, and diabetes prevalence. Higher sNFL levels were significantly associated with increased odds of CVD (adjusted OR = 1.41, 95% CI: 1.05–1.88). The association was significant in non-hypertensive individuals (OR = 2.72, 95% CI: 1.61–4.62) but not in hypertensive individuals (OR = 1.13, 95% CI: 0.81–1.56). sNFL addition to traditional risk models improved predictive accuracy, especially in non-hypertensive individuals (AUC from 0.827 to 0.856). sNFL levels are significantly associated with CVD in the general population, with a strong predictive value in non-hypertensive individuals. Future longitudinal studies should validate sNFL’s efficacy in various populations and explore the underlying mechanisms of its relationship with hypertension and CVD.

Circulating nucleases degrade high-molecular-weight cell-free DNA in coronary vessels during acute myocardial infarction

Abstract Introduction Extracellular cell-free DNA (cfDNA) originates from various biological processes, including cell turnover, apoptosis, necrosis, NETosis, and other forms of cell death. In healthy individuals, highly fragmented cfDNA is elevated in the bloodstream due to infection, cancer, inflammation and thrombosis. In thrombotic vascular diseases, neutrophils release elongated, high-molecular-weight DNA with bioactive and prothrombotic properties in a process termed NETosis. Circulating nucleases are thought to counteract pathogenic properties of NETs. However, attempts to connect total cfDNA concentration to nuclease activity have failed. Purpose We aimed to elucidate the in vivo connection between extracellular DNA and circulating nucleases in thrombotic vascular diseases by investigating DNA length as a potential novel biomarker in cardiovascular disease. Methods Citrated plasma from healthy volunteers (n=363) and from ST-elevation myocardial infarction (MI) coronary aspirates (n=59) and peripheral arterial sites (n=96) were analyzed for DNA concentration, length, and nuclease activity. Total DNA concentration was measured with a fluorescent DNA binding dye, and elongated DNA assessed using qPCR and Agilent bioanalyzer high-sensitivity DNA chip. Nuclease activity was measured by single radial enzyme-diffusion (SRED) assay. Cardiac MRI assessed infarct size acutely and six months after the event. Results Coronary aspirates exhibited an elevation in total DNA, with a significant rise in high-molecular weight cell-free DNA in addition to an increase in sub-nucleosome fragmentation (Figure 1). Peripheral and coronary plasma samples from individuals with myocardial infarction (MI) showed elevated levels of total and elongated cfDNA. When compared to healthy individuals, the integrity of cfDNA was lower in peripheral samples and higher in coronary aspirates. During myocardial infarction, circulating nuclease activity was elevated systemically but low at the coronary culprit lesion site. Elongated high-molecular-weight cfDNA accumulates primarily in coronary vessels but is also elevated systemically. In coronary aspirates, nuclease activity negatively correlated with elongated DNA and DNA integrity (rs=0.6255, p&amp;lt;0.0001). Conclusions Elongated DNA accumulates during MI at the culprit lesion site. Its inverse correlation with nuclease activity underscores the role of nucleases in cleaving elongated DNA to shorter, soluble fragments. This study establishes DNA length and integrity as novel biomarkers in thrombotic cardiovascular disease.Figure 1:Coronary cfDNA lengthFigure 2:Nuclease activity and cfDNA

Red blood cell distribution width to albumin ratio as novel prognostic biomarker in cardiovascular disease

Abstract Background The red blood cell distribution width-to-albumin ratio (RAR) is a novel, cost-effective, and accessible biomarker for inflammation. Although numerous studies have investigated the relationship between RAR values and mortality in cardiovascular diseases (CVD), the generalizability and robustness of these individual studies' results remain uncertain, underscoring the necessity for a meta-analysis. Purpose To explore the pooled association between RAR and all-cause mortality in several CVDs. Methods A systematic search for eligible studies was conducted across scientific databases, including Scopus, Web of Science, PubMed, and ProQuest, up to February 1, 2024. Mortality hazard ratios (HRs) for the highest versus lowest RAR categories were pooled using the Generic Inverse Variance method. A dose-response meta-analysis was performed using generalized least-squares regression to estimate trends of HRs across dose categories. Publication bias was assessed by inspecting funnel plots and performing Egger's test. Subgroup analyses were based on the underlying CVDs. Statistical analyses were executed in RStudio software, with the significance threshold set at p &amp;lt; 0.05. Results This systematic review and meta-analysis reviewed 16 high-quality studies published between 2021 and 2024 with 30,933 participants. The underlying diseases in the included studies were Acute Myocardial Infarction (AMI), Heart Failure (HF), Atrial Fibrillation (AF), aortic valvular disease, stroke and pulmonary embolism. The lowest quartile range for RAR was 0.80–1.20 ml/g, while the highest quartile ranged from 6.09 to 15.45 ml/g. Pooled results with a random-effect model showed that patients with higher RAR had significantly increased risk of all-cause mortality (HR 1.88, 95%CI 1.59–2.23, p&amp;lt;0.0001, I² = 91%). Non-linearity was observed in the dose-response relationship; thus, it was primarily illustrated using the restricted cubic spline model, with the linear model provided for comparison. A 1 ml/g increase in RAR corresponded to a 27% rise in the predicted HR for all-cause mortality (HR 1.27, 95%CI 1.16–1.39; p &amp;lt; 0.0001). Subgroup analyses based on the primary CVD diagnoses in the included studies confirmed a significant association between RAR values and all-cause mortality across all subgroups (AMI: HR 2.43, 95%CI 1.52–3.87, I² = 45%; HF: HR 1.78, 95%CI 1.13–2.81, I² = 94%; Stroke: HR 1.58, 95%CI 0.94–2.67, I² = 90%; Other CVD: HR 1.97, 95%CI 1.34–2.89, I² = 57%; p for interaction 0.24). Conclusion This is the first meta-analysis demonstrating the positive dose-response association between RAR values and all-cause mortality in CVD. Thus, RAR may be considered as a simple and practical prognostic biomarker in CVD.Forest plot based on CVD diagnosisDose-response RAR value to HR

Awareness and perceptions of cardiologists towards systemic inflammation in atherosclerotic cardiovascular disease: A multi-national survey across 5 geographical regional sub-groups

Abstract Background Despite improvement in the management of atherosclerotic cardiovascular disease and chronic kidney disease (ASCVD+CKD), cardiovascular (CV) mortality remains unacceptably high. Evidence suggests that systemic inflammation (SI) is one of the key drivers for disease progression, yet clinical perceptions towards SI amongst cardiologists is poorly understood. Purpose FLAME-ASCVD (systemic inFLAMmation and rolE of hsCRP as a biomarker in AtheroSclerotic CardioVascular Disease) survey assessed cardiologist’s perceptions and awareness towards the role of SI and high sensitivity C-reactive protein (hsCRP) in patients (pts.) with ASCVD+CKD. Overall results were reported earlier1 and here we present results by sub-groups across 5 geographical regions. Methods This was a multinational, cross-sectional, online survey-based study conducted during March-May 2023. Eligible participants were cardiologists (interventional and general) who treated ≥15 ASCVD+CKD pts./month and practiced for ≥3 years. To minimize selection bias, specific study objective was not disclosed until eligibility criteria was met. An online questionnaire was used. Analysis included 5 sub-groups: Europe (EU: France, Germany, Italy, UK), Asia Pacific (AP: Australia, India), East Asia (EA: China, Japan), LatAm (LA: Brazil), Middle East (ME: Saudi Arabia). Descriptive statistics were used. Results Of the 1755 respondents, 602 eligible cardiologists (~60/country) completed the survey across the EU (n=241), AP (n=121), EA (n=120), LA (n=60), and ME (n=60). Hypertension, hyperlipidaemia, and lifestyle (diet/exercise) were consistently amongst the top 3-5 CV risk factors discussed with pts, across all regions. SI as a CV risk factor was discussed consistently less across regions (overall 10th at 43%, ³9th across regions, EU: 30%) (Table 1). Across regions, &amp;gt;70% of cardiologists acknowledged an intention to consider testing for SI, with the lowest being in the EU at 46%. Of those in the EU who did not consider testing, 53% (n=129) cited lack of medication as the reason. Across all regions, SI was consistently perceived as a risk factor for recurrent CV events (³65%) and residual inflammatory risk (47-85%), yet a noticeable proportion (52-92%) cardiologists acknowledged a need to learn more about SI in ASCVD+CKD (highest in ME, 92%). While overall (aided: 47% [n= 571]; unaided: 24%) considered hsCRP test for SI, few variations were observed (EA/AP/EU, aided: 55/52/33 and unaided: 22/24/03) (Table 2). Proven efficacy of hsCRP test, availability of medication for SI, and guideline recommendation were acknowledged as unmet needs to inform clinical care in ASCVD+CKD. Conclusion In this survey across regional sub-groups, discussion of SI as a CV risk factor was consistently low. Barring few noticeable variations in testing patterns in EU, perceptions towards SI were generally consistent. There is a need for better understanding and guidance on SI and hsCRP testing in practice.

Exploring protein biomarkers of physical activity and cardiovascular disease

Abstract Background The beneficial effects of physical activity on cardiovascular risk are well documented. Yet, the underlying mechanisms by which physical activity prevents cardiovascular disease remain unclear (1). Advancements of omics technology enables us to explore novel biomarkers of this complex molecular relationship and potentially unravel underlying molecular mechanisms involved in the progression of cardiovascular events. Purpose To investigate associations between long term physical activity and plasma proteins. As a second step investigate associations between identified plasma proteins and future myocardial infarction and mortality. Methods In the population based prospective cohort Uppsala Longitudinal Study of Adult Men (ULSAM), men were repeatedly investigated with a validated questionnaire on physical activity at ages 50, 60 and 70. 720 plasma proteins were analyzed at age 70 (n=782) with follow up on mortality data for 30 years. In the case-cohort MIMI (Markers of Imminent Myocardial Infarction study) the same plasma proteins were measured in disease-free individuals from six European cohorts. Individuals with acute myocardial infarction within 6 months from baseline were defined as cases (n=420) and up tto four cohort representatives per case were defined as controls (n=1598). Results Higher level of physical activity during 20 years was significantly associated with 12 plasma proteins, in a linear regression model adjusted for age, education and smoking after Bonferroni correction for multiple testing (p&amp;lt;0.000069). After additional adjustment for known cardiovascular risk factors (education, smoking status, BMI, blood pressure, low-density lipoprotein, diabetes diagnosis, and hypertensive-, lipid- or diabetes-treatment), all 12 proteins remained negatively associated with higher level of physical activity in the ULSAM cohort (p&amp;lt;0,05). Two proteins of the 12 proteins associated with physical activity were also significantly associated with future myocardial infarction in the MIMI study (FGF21 and IL6, p&amp;lt;0.05 for both) and 10 proteins were associated with increased mortality risk in the ULSAM cohort (p&amp;lt;0.05 for all). Moreover, proteins that were more negatively associated with physical activity were generally more positively associated with the risk of myocardial infarction in MIMI (Figure). Conclusion Multiple novel associations were found between long-term physical activity and plasma proteins. Some proteins were also associated with future myocardial infarction in an independent cohort which could suggest that the cardioprotective effects of physical activity to some degree may be mediated via the circulating proteome. Our findings encourage additional studies in order to understand the underlying causal mechanisms of these associations.

A direct comparison of non-invasive blood markers of liver fibrosis as associates of cardiovascular risk. The Athens cardiometabolic registry

Abstract Background Non-alcoholic fatty liver disease (NAFLD) is associated with increased risk of cardiovascular disease (CVD). This risk increases further with advanced liver fibrosis. To date, non-invasive tests (NIT) for liver fibrosis risk have not been established as diagnostic or prognostic biomarkers of CVD. Purpose The purpose of the study was to compare NIT of liver fibrosis with respect to their associations with vascular injury and cardiovascular (CV) events. Methods Individuals without clinically overt CVD and patients with CVD (total n=1,692), consecutively recruited in the Athens Cardiometabolic Registry, were analysed in this study. We retrospectively evaluated the association of NIT (i.e. Fibrosis-4 index (FIB-4), NAFLD Fibrosis score (NFS), BARD score) with indices of subclinical arterial injury (i.e. carotid wall thickness and atherosclerotic plaque for subclinical atherosclerosis, pulse wave velocity (PWV), for arterial stiffness) and the incidence of CV events. Patients were followed-up for CV events for a median time of 39 months. Results Elevated FIB-4 (&amp;gt;2.67) was associated with vascular damage [adjusted odds ratio (aOR)=2.97, 95% confidence intervals (CI)= 1.71-5.78 for maximal wall thickness (maxWT) and aOR=4.58, 95% CI=2.44-8.60 for PWV] across the entire study population, including those with or without clinically overt CVD. NFS&amp;gt; 0.675 showed similar associations but lacked significant correlation with arterial stiffness in coronary artery disease patients. FIB-4&amp;gt; 2.67 and BARD&amp;gt; 2 were associated with increased risk of the composite endpoint of CV death, acute myocardial infarction, or coronary revascularization [adjusted hazard ratio, (aHR)=2.00, 95% CI= 1.12-3.55 and aHR=3.36, 95% CI=1.03-11.0 for FIB-4 (n=842) and BARD (n=887) respectively]. FIB-4 and BARD as continuous variables confered incremental prognostic value compared to European Systematic Coronary Risk Evaluation 2 (SCORE2). Conclusion In a population with a wide range of cardiometabolic risk, FIB-4 was associated with both markers of atherosclerotic disease and arterial stiffness as well as with adverse CV events. These data support further investigation of FIB-4 as a biomarker of CV risk.

Inhibition of ceramide de novo synthesis mitigates atherosclerosis

Abstract Background Atherosclerosis is the primary underlying cause of cardiovascular disease. Plasma ceramides are positively correlated with an increased risk of major adverse cardiovascular events and serve as an alternative biomarker for cardiovascular disease. Importantly, ceramide levels are notably higher within atherosclerotic plaques. Therefore, inhibiting the ceramide de novo synthesis pathway in the plaques may serve as a potential therapeutic intervention for atherosclerosis. Purpose We aim to assess the therapeutic efficacy of a nano-formulation of myriocin, a targeted inhibitor of serine-palmitoyl transferase, the key enzyme in ceramide de novo synthesis, in treating atherosclerosis. Methods Myriocin was encapsulated within the hydrophobic core of lipid-based nanoparticles specifically designed for this study. Male and female ApoE-/- mice, subjected to an 8-week high-fat diet regimen, received either myriocin nanoparticles (treatment group) or empty nanoparticles (control group) intravenously once a week for 4 weeks at a dose of 1.3 mg/kg body weight while still on the high-fat diet. At the study endpoint, whole aortas were harvested and subjected to Oil Red O staining to quantify the total lesion area. Histological staining was performed on serial sections of the aortic root to analyze the cellular composition within the plaques. Plasma samples were collected for total cholesterol measurement and lipidomic analysis. Results Upon intravenous administration, myriocin nanoparticles exhibited preferential accumulation in atherosclerotic plaques. Compared to control mice, those treated with myriocin nanoparticles exhibited a 58.7% reduction in total lesion area in male ApoE-/- mice and a 64.6% reduction in female ApoE-/- mice (p&amp;lt;0.0001). Consistently, the lesion areas in the aortic root were also reduced in the treated mice. Notably, myriocin nanoparticles enhanced collagen and alpha-smooth muscle content, while reducing macrophages and neutral lipids within the plaques, leading to a reduced plaque vulnerability index and preventing rupture events. TUNEL staining revealed reduced cellular apoptosis in the atherosclerotic plaques, resulting in a smaller necrotic area in the treated mice. In vitro experiments demonstrated that myriocin nanoparticles effectively inhibited lipopolysaccharide-induced inflammatory response and oxidative stress in bone marrow-derived macrophages. Furthermore, myriocin nanoparticles restored the gene expression of enzymes involved in the fatty acid beta-oxidation pathway, such as ACOX1 and CPT1A, in foam cells. Conclusion Using myriocin nanoparticles to inhibit excessive ceramide production due to inflammation reduces inflammation, and ROS production, while restoring fatty acid beta-oxidation in macrophages. This approach emerges as an effective strategy for targeting ceramide de novo synthesis within atherosclerotic plaques, thereby mitigating atherosclerosis progression and improving plaque stability.