Oriol Alberto Rangel-Zuñiga

Equine Metabolic Syndrome (EMS) is a complex endocrine disorder characterized by obesity, insulin dysregulation (ID), and an increased risk of developing laminitis. Despite growing awareness of EMS, early and accurate diagnosis remains challenging. In this study, we investigated changes in the levels of circulating microRNAs (miRNAs) in plasma samples from horses diagnosed with EMS, compared to obese non-ID horses and healthy controls. Using a targeted real-time PCR OpenArray, we identified four miRNAs —miR-221, miR-186, miR-132, and miR-15a— that were differentially expressed across the groups. Functional enrichment analysis of their validated target genes revealed involvement in key insulin signalling mechanisms, including phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (AKT), AMP-activated protein kinase (AMPK) and forkhead box O protein (FOXO) signalling pathways. Receiver Operating Characteristic (ROC) curve analysis demonstrated that combinations of these miRNAs provided discriminatory power between clinical groups (AUCs ranging from 0.79 to 0.93). These findings support the potential of circulating miRNA levels as non-invasive predictive biomarkers for EMS and related metabolic dysfunctions. Moreover, the signalling pathways identified provide mechanistic insight and establish a foundation for future studies aimed at therapeutic target discovery.

Type 2 diabetes mellitus (T2DM) is a growing global health challenge. Conventional diagnostic tools have limited sensitivity and specificity for early-stage disease. In this context, long non-coding RNAs (lncRNAs) have emerged as promising biomarkers for T2DM risk. However, studies exploring their predictive value remain limited. This study aimed to evaluate the potential of circulating lncRNAs in T2DM development and to assess their interaction with dietary interventions. The study included 462 non-diabetic participants from the CORDIOPREV study, followed for 5 years under Mediterranean or low-fat diet interventions. Expression levels of 22 circulating lncRNAs were quantified by qPCR. A series of statistical and machine learning methods were applied. Random forest analysis identified four lncRNAs (XIST, LINC01116, CASC2, LINC01370) as predictive of T2DM incidence. The combination of these lncRNAs with clinical variables significantly improved prediction performance (AUC = 0.730) compared to models with Hb1Ac (p = 0.015) or clinical variables alone (p < 0.001). Regarding the constructed lncRNA score from the previous model, a lower lncRNA score was associated with a reduced risk of developing T2DM (HR 0.37 (0.24-0.59), p < 0.001), and showed an inverse correlation with the disposition index among individuals following a Mediterranean diet (r = - 0.18, p = 0.009). Circulating lncRNAs, particularly integrated into an epigenetic score, represent promising predictive biomarkers for T2DM development. The interaction with dietary intervention-especially the Mediterranean diet-supports their potential use in guiding personalized dietary strategies for T2DM prevention in high-risk populations.

Background: Familial hypercholesterolemia (FH) is an autosomal dominant disorder associated with a markedly increased risk of premature atherosclerotic cardiovascular disease (ASCVD). Genetic and molecular factors, including microRNAs (miRNAs), may serve as biomarkers to improve disease risk prediction. miRNAs are key regulators of biological processes involved in cardiovascular pathology. This study aimed to identify miRNAs that, combined with clinical variables, enhance ASCVD prediction in asymptomatic FH patients from the SAFEHEART cohort. Methods: A total of 400 subjects diagnosed with FH were included. Coronary calcium score (Agatston score) was measured by computed tomography and stratified into three risk categories (0–99, 100–299, >300). Circulating plasma miRNA levels were quantified using the OpenArray platform. Statistical analyses characterized baseline features and assessed predictive performance using ROC curve analysis. Functional enrichment was explored through the MIENTURNET database. Results: Individuals carrying the null allele for the LDL-R gene showed significantly higher plasma miR-1 levels than those carrying the defective allele (p=0.042). Likewise, subjects with an Agatston score of 0–99 exhibited higher miR-1 levels than those with scores >300 (p=0.025). A predictive model based on clinical variables including age, sex, hypertension, smoking, BMI, LDL burden, and lipoprotein(a) achieved an AUC of 0.846 for ASCVD prediction. Incorporating miR-1 increased the AUC to 0.878, representing a statistically significant improvement (DeLong test, p=0.005). Conclusions: miR-1 emerged as a promising biomarker for ASCVD risk prediction in FH patients. When integrated with conventional clinical variables, miR-1 improved the model’s predictive accuracy. These findings suggest that miR-1 could serve as a useful molecular marker for enhanced cardiovascular risk stratification in asymptomatic individuals with familial hypercholesterolemia.

Background Recent evidence links diet and physical activity with type 2 diabetes mellitus (T2DM) remission, but emerging findings suggest that immune system dysregulation may play a crucial role. This study aimed to investigate the associations between neutrophils and T2DM remission. Methods We conducted a comprehensive analysis of newly-diagnosed T2DM patients (N = 183) from the CORDIOPREV study, without glucose-lowering treatment, and were randomized to follow either a Mediterranean or low-fat diet. Patients were classified into two groups: Responders, who achieved T2DM remission (n = 73), and Non-Responders, who did not achieve remission during the 5-year dietary intervention (n = 110). Neutrophil count and their related-ratio (NER, NBR, NLR and NHR, normalized with erythrocytes, basophils, lymphocytes, and HDL respectively) were measured at the baseline and 5 years of follow-up. Results The lowest baseline tertile of neutrophil count was associated with an increased likelihood of T2DM remission among patients following a Mediterranean diet (but not for low-fat diet) when compared with the highest tertile [adjusted HR of 4.23 (95% CI: 1.53–11.69)], in which similar results were observed for NER and NHR. When considering clinical and neutrophil variables, the predictive capacity of this model yielded an AUC of 0.783 (95% CI: 0.680–0.822). Furthermore, after 5-years, Responders exhibited lower neutrophil count compared to Non-responders (p = 0.006) and a significant decrease in neutrophil count (p = 0.001) compared to baseline. This decrease in neutrophil count in Responders who consumed a Mediterranean diet exhibited a significant increase in Insulin Sensitivity and Disposition Index (p = 0.011 and p = 0.018) after the follow-up period. Conclusion These findings suggest that neutrophil count can help in identifying patients that are more likely to achieve T2DM remission following a Mediterranean diet, suggesting a role on insulin sensitivity and β-cell function. Further research holds promise for providing valuable insights into the pathophysiology of T2DM. Clinical trial Registration ID: NCT00924937; URL Clinical trial: https://clinicaltrials.gov/study/NCT00924937?cond=NCT00924937&rank=1.

Background Type 2 diabetes mellitus (T2DM) is currently a major global public health problem. Although disease remission is possible, few biomarkers have been identified which can help us select the diet that best promotes remission. Our aim was to study the potential of miRNAs as a tool to apply the Mediterranean diet or the low-fat diet in order to achieve T2DM remission in patients with coronary heart disease. Methods From the CORDIOPREV study (n = 1002), a prospective, randomized, single-blind, controlled dietary intervention trial, all patients newly diagnosed with T2DM (n = 190) at baseline were included in the present study. Of these, after adhering to a low fat or Mediterranean diet for 60 months, 73 patients showed T2DM remission (Responders) and 110 continued with the disease (Non-responders). Plasma levels of 56 miRNAs were determined by RT-PCR. Generalized linear model, ROC curves and COX regression analyses were performed. Results We observed that patients with low baseline plasma levels of miR-let7b-3p showed a high probability of T2DM remission after the consumption of a low-fat diet. In addition, patients with high levels of miR-141-5p, miR-182, and miR-192 at baseline showed a high probability of T2DM remission after following the Mediterranean diet. Scores built using miRNAs and clinical variables showed that high levels of a low-fat diet score and a high Mediterranean diet score were associated with a high probability of T2DM remission. Conclusion MiRNAs could be used as a tool for selecting the most efficient nutritional therapy (mediterranean or low-fat diet) to achieve T2DM remission in patients with coronary heart disease.

The incidence of type 2 diabetes mellitus (T2DM) is growing in Western countries. Nutritional interventions that promote high-quality dietary patterns could help reverse this trend. We aimed to evaluate whether changes in Nutrient-Rich Food Index 9.3 (NRF9.3) were related to the risk of developing T2DM in patients with coronary heart disease (CHD). The study was carried out in the context of two healthy dietary interventions (a Mediterranean and a low-fat diet). For this purpose, we evaluated all the patients in the CORDIOPREV study without T2DM at baseline. Data were obtained during the first 5 years of dietary intervention. The score was calculated using the Food Frequency Questionnaires at baseline and after 1 year of intervention. After 5 years of follow-up, 106 patients developed T2DM (incident-T2DM), while 316 subjects did not (non-T2DM). Total NRF9.3 score and changes during the first year of intervention were compared between incident-T2DM and non-T2DM. Incident-T2DM showed less improvement in NRF9.3 than non-T2DM (p = 0.010). In the multi-adjusted Cox proportional hazard study, patients with greater improvement in NRF9.3 had over 50% less risk of developing T2DM compared with the lowest tertile (HR 2.10, 95%, CI = 1.12–3.56). In conclusion, improved diet quality in terms of nutrient density after the dietary intervention was associated with a lower risk of T2DM in patients with CHD.