Águeda González-Rodríguez

Bone morphogenetic protein 12 (BMP12) is known to regulate mesenchymal differentiation and extracellular matrix remodeling. Given its structural and signaling similarities to BMP2, which promotes osteogenic phenotypic switching and pro-atherogenic responses in vascular smooth muscle cells (VSMCs), BMP12 may also contribute to vascular remodeling and atherosclerosis progression. This translational study aimed to unravel BMP12 involvement in cardiovascular disease (CVD). BMP12 effects on atherogenesis-related processes in VSMCs in absence or presence of BMP type I receptor inhibitors (DMH2 and LDN-193189) were evaluated. Additionally, circulating BMP12 levels were measured in a well-characterized cohort of patients with CV risk factors and correlated with cardiometabolic disorders and vascular damage markers. Results from the experimental study showed increased migration and calcification in BMP12-treated VSMCs. Mechanistically, BMP12 induced expression of osteogenic markers (RUNX2 and ALP) and genes related to cellular migration ( Mmp9 ), while reducing contractile marker Desmin . These pro-atherogenic effects were blocked using BMP type I receptor inhibitors. Furthermore, the clinical study revealed higher serum BMP12 levels in patients with CVD and type 2 diabetes. Moreover, circulating BMP12 concentrations were elevated in patients with carotid and femoral plaques, correlating with plaque volume. Notably, BMP12 emerged as an independent predictor of femoral atherosclerosis. Collectively, these findings identify BMP12 as a previously unrecognized mediator of vascular remodeling, promoting pro-atherogenic effects in VSMCs through canonical BMP signaling. The association between elevated circulating BMP12 levels and subclinical vascular damage further supports its potential value as a contributor to atherosclerosis progression and as a promising novel candidate biomarker for CV risk.

Traumatic brain injury (TBI) is a highly heterogeneous disease and achieving an accurate diagnosis remains a significant challenge. Biomarkers play a crucial role in minimizing the reliance on invasive techniques like computed tomography, which also have significant economic costs. Human samples were obtained from prospective cohort studies. Mice were subjected to an experimental model of traumatic brain injury. Biomarker levels, gene expression, and blood–brain barrier integrity were analyzed using ELISA, qRT-PCR, and Evans Blue assay; data were statistically evaluated using parametric or non-parametric tests as appropriate. This study focuses on evaluating the role of matricellular protein thrombospondin-1 (TSP-1) and the tight junction proteins occludin and ZO-1 as potential biomarkers of TBI. We showed that lower serum TSP-1 levels correlated with poor patient outcomes at 6 months compared to those patients with a good outcome. Additionally, the disruption of the blood–brain barrier (BBB) and subsequent release of tight junction proteins allowed us to identify occludin as a potential biomarker for prognosis in a cohort of TBI patients and as a diagnosis biomarker in a subgroup of patients with mild TBI, but its discriminative power as a diagnosis biomarker appears modest, as reflected by an AUC of 0.693. On the other hand, ZO-1 exhibited increased levels but limited diagnostic utility. These findings highlight the critical role of TSP-1 in maintaining BBB integrity and regulating the inflammatory response after a TBI, supported by the worsened condition observed in TSP-1-deficient animals. These results demonstrate the potential of TSP-1 and occludin as valuable biomarkers for secondary injury and disease progression in patients with mild to moderate/severe TBI.

Liver fibrosis represents the main risk factor not only for liver-related but also for overall mortality in metabolic dysfunction-associated steatotic liver disease (MASLD) patients, being metabolic dysfunction-associated steatohepatitis (MASH) its more severe clinical form. We recently developed a non-invasive algorithm termed BMP8A Fibrosis Score (BFS) which is able to identify MASH patients with advanced liver fibrosis. The aim of this study was to validate the BFS comparing its diagnostic accuracy with that of other scoring systems developed to assess liver fibrosis in MASH patients. Serum BMP8A was measured in 302 patients with biopsy-proven MASH: 171 with non- or mild fibrosis (F0-F2) and 131 with advanced fibrosis (F3-F4) recruited from seven university hospitals located in different cities in Spain. BFS, Fibrosis-4 (FIB-4) Index, NAFLD Fibrosis Score (NFS), Hepamet Fibrosis Score (HFS), and AST-to-Platelet Ratio Index (APRI) were calculated for each patient. The diagnostic accuracy of the scoring systems was determined according to the area under the receiver operating characteristic (AUROC) curve, sensitivity, specificity, positive (PPV) and negative (NPV) predictive values, and likelihood ratios (LR). BFS showed higher overall accuracy than the other liver fibrosis algorithms calculated in the study cohort, presenting an AUROC of 0.750 for predicting advanced liver fibrosis (F3-F4), and correctly classifying 70.9% of F3-F4 patients with a sensitivity of 58.0%, a specificity of 80.7%, a 71.5% NPV, a 69.7% PPV, a 3.0 LR+, and a 0.5 LR-; the other predictive scores correctly classified a lower percentage of these patients (63.6% for FIB-4 ≥ 2.67, 63.2% for HFS ≥ 0.47, 57.3% for APRI ≥ 1.5 and 56.9% for NFS ≥ 0.675). BFS eliminates the grey area as it uses a single cut-off value (0.46), which is its key advantage over the others, reducing the number of patients with undetermined results (43.4% for FIB-4, 39.1% APRI, 37.4% for HFS, and 24.1% NFS). In sum, BFS properly classified more patients with advanced liver fibrosis (F3-F4) than the other scoring systems, eliminating indeterminate results and improving risk stratification.

Background: Traumatic brain injury (TBI) is a highly heterogeneous disease and achieving an accurate diagnosis remains a significant challenge. Biomarkers play a crucial role in minimizing the reliance on invasive techniques like computed tomography, which also have significant economic costs. Methods: Human samples were obtained from prospective cohort studies. Mice were subjected to an experimental model of traumatic brain injury. Biomarker levels, gene expression, and blood-brain barrier integrity were analyzed using ELISA, qRT-PCR, and Evans Blue assay; data were statistically evaluated using parametric or non-parametric tests as appropriate. Results: This work focuses on evaluating the role of matricellular protein thrombospondin-1 (TSP-1) and the tight junction proteins occludin and ZO-1 as potential biomarkers of TBI. We showed that lower serum TSP-1 levels correlated with poor patient outcome at 6 months compared to those patients with a good outcome. Additionally, the disruption of the blood-brain barrier (BBB) and subsequent release of tight junction proteins allowed us to identify occludin as a potential biomarker for prognosis in a cohort of TBI patients and as a diagnosis biomarker in a subgroup of patients with mild TBI. Conclusions: These findings highlight the critical role of TSP-1 in maintaining the BBB integrity and regulating the inflammatory response after TBI, supported by the worsened condition observed in TSP-1-deficient animals. These results demonstrate the potential of TSP-1 and occludin as valuable biomarkers for secondary injury and disease progression in patients with mild to moderate-severe TBI.

Obstructive sleep apnea (OSA), characterized by intermittent hypoxia (IH), is strongly associated with metabolic syndrome and metabolic dysfunction-associated steatotic liver disease (MASLD). IH exacerbates MASLD progression through oxidative stress, inflammation, and lipid accumulation. This study aims to investigate the impact of oxygen normalization on metabolic dysfunction in OSA patients using continuous positive airway pressure (CPAP) therapy, and in mice exposed to IH followed by a reoxygenation period. In the clinical study, 76 participants (44 OSA patients and 32 controls) were analyzed. OSA patients had higher insulin resistance, triglycerides, very low density lipoprotein (VLDL) content, and liver enzyme levels, along with a higher prevalence of liver steatosis. After 18 months of CPAP therapy, OSA patients showed significant improvements in insulin resistance, lipid profiles (total cholesterol and VLDL), liver function markers (AST and albumin), and steatosis risk scores (Fatty Liver Index and OWLiver test). In the experimental study, IH induced hepatic lipid accumulation, oxidative stress, and inflammation, and reoxygenation reversed these deleterious effects in mice. At the molecular level, IH downregulated fatty acid oxidation (FAO)-related genes, thus impairing the FAO process. Reoxygenation maintained elevated levels of lipogenic genes but restored FAO gene expression and activity, suggesting enhanced lipid clearance despite ongoing lipogenesis. Indeed, serum β hydroxybutyrate, a key marker of hepatic FAO in patients, was impaired in OSA patients but normalized after CPAP therapy, supporting improved FAO function. CPAP therapy improves lipid profiles, liver function, and MASLD progression in OSA patients. Experimental findings highlight the therapeutic potential of oxygen normalization in reversing IH-induced liver damage by FAO pathway restoration, indicating a metabolic reprogramming in the liver.

Given the lack of accurate diagnostic methods of acetaminophen (APAP)-induced acute liver failure (ALF), the search for new biomarkers for its diagnosis is an urgent need. The aim of this study was to evaluate the role of bone morphogenetic protein 6 (BMP6) in APAP-induced ALF progression and its potential value as a biomarker of ALF. Hepatic and circulating BMP6 expression was assessed in APAP-treated mice and in serum samples from patients with APAP overdose. In addition, BMP6 expression and release was evaluated in hepatocytes after APAP exposure. BMP6 gene was silenced in Huh7 cells prior to APAP treatment and the culture medium (CM) was added to THP1 cells to evaluate the paracrine effects of hepatocyte BMP6 on APAP toxicity. Hepatic and serum BMP6 levels were increased in mice after APAP-induced ALF. In addition, a positive correlation was observed between circulating BMP6 and ALT activity in patients exposed to APAP overdose. Moreover, hepatocytes expressed and released BMP6 to the CM after APAP treatment. Indeed, the CM from APAP-treated Huh7 cells upregulated M1 and M2 markers in THP1 monocytes. The CM from BMP6-silenced Huh7, which was depleted of BMP6, reduced the expression of M2 markers in THP1 cells. In fact, expression of M2 markers was increased in THP1 cells exposed to BMP6. This study reveals that hepatic BMP6 expression is increased in APAP-induced acute liver injury, positioning it as a potential new biomarker of liver damage severity. Moreover, our data indicate that BMP6 might play a role in the hepatocyte–macrophage crosstalk during APAP-induced hepatotoxicity.

Aim: To determine the prevalence of skeletal muscle alterations (SMA) in patients with meta-bolic dysfunction-associated steatotic liver disease (MASLD) and to assess the significance of circulating myokines as potential biomarkers of SMA in patients with MASLD. Material and methods: Measurements of both skeletal muscle strength and skeletal muscle mass parameters, anthropometric and metabolic features as well as serum levels of different myokines were per-formed in a cohort of 62 MASLD patients and then compared according to the stage of liver fi-brosis and hepatosteatosis degree both diagnosed by transitional elastography. Results: No sig-nificant differences were found in skeletal muscle strength and skeletal muscle mass in MASLD patients stratified according to the stage of liver fibrosis. Noteworthy, serum levels of FGF21 were significantly higher in MASLD patients with advanced hepatic fibrosis (F3-F4) than in those with lower stages of hepatic fibrosis (F0-F2) (197.49±198.27 pg/ml vs 95.62±83.67 pg/ml, p=0.049, respectively). MASLD patients with severe hepatosteatosis (S3) had significantly higher serum levels of irisin (1116.87±1161.86 pg/ml) than those with lower grades (S1-S2) (385.21±375.98 pg/ml, p=0.001). Conclusion: SMA were uncommon in our MASLD patients, particularly in those with lower stages of liver fibrosis. An increase in serum FGF21 was detected in patients with a higher stage of fibrosis with possible therapeutic implications in MASLD.

Background and Aims: Cyclooxygenase‐2 (COX‐2) is involved in different liver diseases, but little is known about the significance of COX‐2 in cholestatic injury. This study was designed to elucidate the role of COX‐2 expression in hepatocytes during the pathogenesis of obstructive cholestasis.Methods: We used genetically modified mice constitutively expressing human COX‐2 in hepatocytes. Transgenic mice (hCOX‐2‐Tg) and their wild‐type (Wt) littermates were either subjected to a mid‐abdominal laparotomy or common bile duct ligation (BDL) for 2 or 5 days. Then, we explored the mechanisms underlying the role of COX‐2 and its derived prostaglandins in liver function, and the synthesis and excretion of bile acids (BA) in response to cholestatic liver injury.Results: After BDL, hCOX‐2‐Tg mice showed lower grades of hepatic necrosis and inflammation than Wt mice, in part by a reduced hepatic neutrophil recruitment associated with lower mRNA levels of pro‐inflammatory cytokines. Furthermore, hCOX‐2‐Tg mice displayed a differential metabolic pattern of BA synthesis that led to an improved clearance after BDL‐induced accumulation. In addition, an enhanced response to the BDL‐induced oxidative stress and hepatic apoptosis was observed. In vitro experiments using hepatic cells that stably express hCOX‐2 confirmed the cytoprotective role of prostaglandin E₂ against BA toxicity.Conclusions: Taken together, our data indicate that constitutive expression of COX‐2 in hepatocytes ameliorates cholestatic liver injury in mice by reducing inflammation and cell damage and by modulating BA metabolism, pointing to a role for COX‐2 as a defensive response against cholestasis‐derived BA accumulation and injury.