bluuebunny/crossref_metadata_2025_split huggingface dataset:
Peroxisome proliferator-activated receptor gamma regulates interleukin-6 induced lipoprotein(a) gene expression in human HepG2 cells
Authors: Harb T , Ziogos E , Amat-Alarcon N , Gerstenblith G , Arvanitis M , Leucker T M 
Published: October 2024

Abstract

Background
Lipoprotein(a)[Lp(a)] is an independent risk factor for atherosclerotic cardiovascular disease (ASCVD) and outcomes. The LPA gene promoter contains an interleukin 6 (IL-6) responsive binding site. IL-6 is a pro-inflammatory cytokine and data from patients with inflammatory diseases indicate an impact of the inflammatory milieu on LPA gene regulation and Lp(a) production. In a clinical study, the peroxisome proliferator-activated receptor gamma (PPARγ) agonist pioglitazone was associated with decreased Lp(a) levels. Publicly accessible ChIP-sequencing of human hepatocytes and measures of Lp(a) production by HepG2 cells provide an opportunity to explore PPARγ regulation of hepatic Lp(a) production.

Purpose
To investigate the role of PPARγ as a regulator of hepatic Lp(a) production.

Methods
Public ChIP-sequencing of human hepatocytes was analyzed. Commercially available HepG2 cells were incubated with one of the following: (1) scrambled small interfering RNA (siRNA) alone (control); (2) IL-6 (20ng/mL) and scrambled siRNA ; (3) IL-6 and PPARγ siRNA ; and (4) IL-6 and pioglitazone (25uM). LPA transcription was measured using the quantitative polymerase chain reaction.

Results
Analysis of the ChIP-sequencing data demonstrated that the LPA locus variant rs56393506, which is associated with increased Lp(a) levels and ASCVD risk in the general population, was within the location of the PPARγ binding site in the LPA promoter (Figure 1). IL-6 significantly increased LPA transcription in the HepG2 cells, compared to control (1.65±0.44 vs. 0.66±0.45, p=0.0027). In the presence of siRNA-mediated knock-down of PPARγ (58±17% PPARγ reduction, p&lt;0.0001 vs. scrambled control), the effect of IL-6 on LPA transcription was significantly amplified (6.78±2.4 vs. 1.65±0.44, p=0.0009). IL-6 did not increase LPA transcription in the presence of the PPARγ agonist pioglitazone compared to control (0.16±0.03 vs. 0.66+0.45, p=0.11, Figure 2).

Conclusion
PPARγ is a negative regulator of IL-6 induced hepatic Lp(a) production and may represent a new therapeutic target in patients with inflammatory conditions.Figure 1Figure 2
Single-cell mass cytometry analysis unveils prothrombotic expression and heterogeneity in essential thrombocythemia
Authors: Kuehne S G , Dill V , Kirmes K , Han J , Klug M , Bloxham C J , Raake P W J , Bernlochner I , Bongiovanni D 
Published: October 2024

Abstract

Background/Introduction
Essential thrombocythemia (ET) is recognized for its hallmark elevated platelet count and a consequent increased risk of thromboembolic events including coronary events. Despite its clinical significance, the pathophysiological mechanisms predisposing ET patients to thrombosis remain largely undefined.

Purpose
The aim of this pioneering study was to shed light on the increased thrombotic risk in ET by conducting a detailed phenotypical characterization of platelet expression using single-cell mass cytometry, comparing ET patients to age- and sex-matched healthy controls.

Methods
Platelet samples from six patients diagnosed with ET and six healthy individuals were analyzed. This analysis utilized an extensive panel of 18 transmembrane regulators pivotal for platelet function and activation. Measurements were taken both at baseline and after ex-vivo stimulation with thrombin receptor-activating peptide (TRAP). Advanced clustering analysis via the FlowSOM algorithm helped identify specific subclusters of platelets with a prothrombotic expression profile.

Results
Our investigation revealed a significant overexpression of the activation marker CD62P (P-Selectin, p=0.049) and of the collagen receptor GPVI (p=0.044) in non-stimulated ET platelets (Figure 1A-C). Conversely, a notable decrease in expression was observed for the fibrinogen receptor GPIIb/IIIa units CD41 (p=0.036) and CD61 (p=0.044), as well as for the von Willebrand factor receptor CD42b (p=0.044). The FlowSOM analysis distinguished two prothrombotic subclusters of ET platelets, particularly highlighting one cluster, which was significantly overrepresented in ET (22.13% of total ET platelets versus 2.94% in controls, p=0.035, Figure 1D-E).

Conclusion
This study presents compelling evidence of a distinct prothrombotic phenotype in ET, marked by significant alterations in platelet expression profiles. The overexpression of CD62P and GPVI, alongside the underexpression of CD41, CD61, and CD42b, underscores the complex pathophysiological landscape of ET. Importantly, we identified a specific subcluster, particularly overrepresented in ET, with a specific prothrombic signature. Our findings, while only hypothesis generating, propose GPVI as a potential therapeutic target to mitigate thrombosis in ET patients, paving the way for targeted interventions in this high-risk population.
"Honey pot"-like lesion formation: impact of catheter contact angle on lesion formation by novel temperature controlled ablation catheter in a porcine experimental model
Authors: Ogiso S , Fukaya H , Ogawa E , Mori H , Masuda Y , Yazaki Y , Murayama Y , Saito D , Kobayashi S , Nakamura H , Ishizue N , Kishihara J , Niwano S , Oikawa J , Ako J 
Published: October 2024

Abstract

Background
The DiamondTemp(DT) catheter enables precise temperature-controlled ablation. However, the effects of the DT catheter's contact angle on lesion formation are poorly understood.

Purpose
This study evaluated lesion formation using the DT catheter at different angles in a porcine experimental model.

Methods
We used freshly sacrificed porcine hearts, and radiofrequency catheter ablation with DT was performed at 50 watts for 15 seconds at an upper-temperature setting of 60°C. The contact force (5, 10, and 30 grams) and catheter contact angles (30°, 45°, and 90°) were changed in each set (n = 13 each). The surface width (SW), maximum lesion width (MW), lesion depth (LD), surface area (SA), and distance from the distal edge to the widest area (DW) were evaluated. In addition, the generator impedance drop during radiofrequency application was analyzed.

Results
Figure 1 shows the summarized data in this study. SW and MW were longer at 30° than at 90° (SW 5 g: 6.34 ± 0.91 vs. 5.07 ± 0.87 mm; SW 10 g: 7.46 ± 0.55. vs. 5.60 ± 0.84 mm; SW 30 g: 7.42 ± 0.74. vs. 6.14 ± 0.77 mm; MW 10 g: 8.81 ± 0.55. vs. 7.89 ± 0.84 mm; MW 30 g: 9.24 ± 0.67. vs. 8.03 ± 1.04 mm, p &lt;0.05, respectively). There were no significant differences in LD by catheter angle except at 30 g. SA was larger at 30° than at 90° (SA 5 g: 30.03 ± 0.91. vs. 23.61 ± 0.87; SA 10 g: 29.24 ± 0.55. vs. 21.27 ± 0.84; SA 30 g: 35.86 ± 0.74. vs. 31.74 ± 0.77, p &lt;0.05, respectively). Besides, DW was longer at 30° than at 90° (DW 5 g: 4.28 ± 1.17. vs. 2.50 ± 0.32 mm; DW 10 g: 4.94 ± 0.52. vs. 2.47 ± 0.38 mm; DW 30 g: 5.34 ± 0.26. vs. 2.96 ± 0.40 mm, p &lt;0.05, respectively). There were no significant differences in the impedance drop according to the catheter angle.

Conclusion
With the DT catheters, the lesion width increased at a shallower contact angle, whereas the LD did not. SA also increased at a shallower contact angle. The DT catheter created a large ablation lesion on the proximal side of the catheter, which looked like a "honey pot." (Figure 2)summarized dataImage of honey pot-like lesion formation
Linking antibody against apolipoprotein A-1 to metabolic dysfunction-associated steatohepatitis in mice
Authors: Pagano S , Somm E , Juillard C , Ino F , Ferrari J , Braunersreuther V , Jornayvaz F , Vuilleumier N 
Published: October 2024

Abstract

Background
Metabolic dysfunction-associated fatty liver disease (MASLD) is a common liver condition associated with increased cardiovascular disease (CVD) risk. Cytokeratin 18 (CK-18) is indicative of liver injury and used as a surrogate biomarker covering MASLD to cirrhosis phenotypes. Autoantibodies against apolipoprotein A-1 (AAA-1) are known CVD risk factors inducing MASLD in vitro, but their role in modulating steatohepatitis and fibrosis in vivo is still elusive. We investigated AAA-1's contribution to systemic low-grade inflammation, liver steatosis, and fibrosis using a MASLD mouse model and a validated passive immunization protocol (PIP).

Methods
10 weeks-old male C57BL/6J mice were fed a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) and immunized with AAA-1 (200ug) or control antibodies for ten days. At sacrifice, plasma samples were collected, and CK-18 levels, and proinflammatory cytokines were measured using the Mesoscale Discovery platform. Hematoxylin and Eosin and Sirius Red Fast Green staining were used to reveal liver steatosis and fibrosis, respectively. RNA were extracted from mouse liver samples and mRNA were then analyzed using a commercial fibrosis-specific genes panel (nCounter Human Fibrosis V2 Panel) and the nCounter Analysis System.

Results
Compared to control IgG recipients, CDAHFD mice injected with AAA-1 exhibited significantly elevated plasma levels of CK-18 (5.3 vs 2.1, p=0.031), IL-6 (13 vs 6.9, p=0.035), IL-10 (27.3 vs 9.8, p=0.007) and TNF-α (32.1 vs 24.2, p=0.032), and liver steatosis (93.4% vs 73.8%, p=0.007). Nanostring technology experiments revealed upregulation of several pro-fibrotic m-RNAs in liver tissue from AAA-1 immunized mice compared to the IgG immunized control group. These included Serum Amyloid A (Fold Change (FC) AAA-1 vs IgG : 2.5, p=0.02) , G-protein coupled receptors 65 (FC: 2.7, p=0.01), Periostin (FC: 2.6, p=0.001), Phospholipid transfer protein (FC: 2.2, p=0.0004), Vascular Cell Adhesion Molecule 1 (FC: 2.5, p=0.02) and Angiopoietin-like 4 (FC: 2.8, p=0.01). Histologically, the amount of liver fibrosis remained unaffected, probably due to the short time of the study.

Conclusions
Short AAA-1 PIP induced enhanced liver steatosis and inflammation accompanied by an elevation of the expression of several pro-fibrotic genes in CDAHFD mice, suggesting that AAA-1 may contribute to the transition from MASLD to MASH. Knowing whether increased exposure time to AAA-1 could lead to end stage disease (including cirrhosis and HCC), and if this could apply to humans as well warrant further investigations.
C-C motif chemokine ligand 3 is associated with atherosclerotic plaque vulnerability and increased risk of mortality
Authors: Li Y , Sun J , Persson A , Nitulescu M , Sundius L , Nilsson J , Edsfeldt A , Goncalves I 
Published: October 2024

Abstract

Background and Purpose
Chemokines play a crucial role in the inflammatory process underlying atherosclerotic plaque formation, progression and subsequent cardiovascular events. However, the specific role of C-C motif chemokine ligand 3 (CCL3) in atherosclerosis remains largely unexplored. This study aims to explore associations between intra-plaque levels of CCL3 and plaque vulnerability as well as to investigate the prognostic role of circulating CCL3 levels in patients undergoing carotid endarterectomy.

Methods
The CCL3 protein expression was measured using a proximity extension assay in 207 human carotid plaques and 558 plasma samples from patients undergoing carotid endarterectomy, all enrolled in the Carotid Plaque Imaging Project (CPIP) cohort. Protein expression is quantified as normalized protein expression (NPX), an arbitrary unit on a log2-scale. Plaque components and extracellular matrix including smooth muscle cells, macrophages, neutral lipids, intraplaque hemorrhage, collagen (by histology), and matrix metalloproteinases (assessed biochemically), were quantified. Plaque vulnerability index was calculated as the sum percentage of plaque areas stained for lipids, macrophages and hemorrhage, divided by the sum percentage of smooth muscle cells and collagen. Patients were followed up to 10.2 years to record cardiovascular events and mortality.

Results
Plaque and circulating levels of CCL3 were higher in symptomatic patients compared to asymptomatic patients (plaque, median NPX (interquartile range, IQR): 18.2 (17.4–19.1) vs 17.2 (16.4–18.0), P&lt;0.001; plasma, median NPX (IQR): 1.78 (1.42–2.21) vs 1.56 (1.28–1.97), P&lt;0.001). Plaque levels of CCL3 showed positive correlations with neutral lipids, macrophages, intraplaque hemorrhage, plaque vulnerability index and matrix metalloproteinases including MMP1, MMP2, MMP9, MMP10 and TIMP1, but inversely with smooth muscle cells. Among 511 patients with complete follow-up and circulating CCL3 measures, 144 (28.2%) patients developed cardiovascular events and 123 (24.1%) patients died. Elevated circulating CCL3 levels were associated with increased risk of cardiovascular events (hazard ratio (HR) per standard deviation (SD) = 1.40, 95% CI 1.06–1.84) and mortality (HR per SD=1.77, 95% CI 1.34–2.33), respectively. Higher levels of circulating CCL3 remained independently associated with an increased risk of mortality (HR per SD=1.50, 95% CI 1.09–2.04) even after adjustment for potential confounders.

Conclusions
Plaque levels of CCL3 are associated with plaque vulnerability features. Circulating levels of CCL3 predict higher risk of cardiovascular events and mortality among carotid endarterectomy patients.
Pericardial adipose tissue (PAT) as a cell model to assess patient responsiveness to docosahexaenoic acid (DHA) and eicopentaenoic acid (EPA)
Authors: Quarta S , Santarpino G , Cardetta F , Carluccio M A , Calabriso N , Madonna R , De Caterina R , Massaro M 
Published: October 2024

Abstract

Background
The abundance of epicardial (EAT) and pericardial adipose tissue (PAT) is associated with several cardiac risk factors and a higher carotid intima-media thickness, with a potential contributory role to the development of atherosclerosis and coronary artery disease (CAD). Under conditions promoting obesity, both epicardial and pericardial adipose tissue undergo a metabolic shift, acquiring an inflammatory and pro-atherogenic phenotype. Addressing dysmetabolism and inflammation associated with EAT and PAT are a promising and innovative therapeutic approach for preventing coronary atherosclerosis.
Recent data have highlighted a noteworthy association between the abundance of adipose tissue n-3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), as well as their ratio to the omega-6 FA arachidonic acid (AA), with a lower risk of myocardial infarction.

Purpose
To develop an efficient method for isolation of adipocytes from human PAT and evaluation of adipocyte response to DHA and EPA.

Methods
PAT samples were collected from 9 CAD patients, undergoing coronary artery bypass graft surgery and immediately processed before to assess more than 50 experimental conditions. Isolated adipocytes were morphologically and molecularly characterized. Cell responsiveness was evaluated by cell exposure to inflammatory stimuli, with or without variable amounts of DHA, EPA, or AA assessing a spectrum of gene expressions and pro-inflammatory functional activities.

Results
Pure cultures of pericardial adipocytes exhibited prolonged viability (beyond 72 hours). and retained responsiveness to pro-inflammatory and lipolytic stimuli, while also showing the capacity to activate the insulin signalling pathway. Cell exposure to DHA or EPA resulted in a downregulation of messenger (m)RNA expression for monocyte chemoattractant protein (MCP)-1, interleukin (IL)-6 and metalloproteinase (MMP)-9 (p&lt;0.05) and in upregulation of uncoupling protein-(UCP)-1, UCP-2 and peroxisome proliferator-activated receptor (PPAR)γ (p&lt; 0.05) (Figure 1).
In line with mRNA expression, a significant reduction in both cytokines and MMP-9 protein release was observed (p&lt;0.05) (Figure 2). These effects paralleled a significant incorporation of DHA and EPA into total cell lipids (p&lt;0.05). Functionally, the cell supernatant from DHA- or EPA-conditioned cells displayed diminished ability to attract monocytes in chemotaxis assays. AA exhibited no such activities.

Conclusion(s)
In a novel cellular model of isolated and cultured adipocytes from PAT, DHA and EPA attenuated the inflammatory and dysmetabolic characteristics of this cardiac adipose tissue.
Bidirectional roles of extracellular and intracellular HMGB2 in the development of atherosclerosis
Authors: Wang X Q , Chen J W , Chen C D , Lu L 
Published: October 2024

Abstract
Rationale: HMGB family plays an important role in homeostasis of immunity and regulation of inflammation. However, the role of HMGB2 in atherosclerosis remains uncharacterized.

Objective
The current study aimed to ascertain the relation of HMGB2 levels in macrophages of atherosclerotic plaques and in circulatory monocytes/macrophages with coronary artery disease (CAD), and to investigate the influence of HMGB2 on atherogenesis in mice and inflammatory reaction in vitro.

Methods/Results
We evaluated HMGB2 levels in macrophages of atherosclerotic plaques in coronary endarterectomy tissue from patients with angiographically documented severe coronary artery disease (CAD)(n=23), and in coronary artery specimens from autopsy cases with no or mild atherosclerosis (n=11) by immunohistochemistry. HMGB2 levels were significantly decreased in macrophages of progressive atherosclerotic plaques, as compared with those from mild plaques. Moreover, we also analyzed HMGB2 levels in peripheral blood monocytes from patients with CAD (n=44) or age- and sex-matched non-CAD controls (n=44). The results showed that decreased intracellular HMGB2 protein levels in monocytes were associated with CAD. In experiments, HMGB2 deficiency significantly promoted atherosclerosis in ApoE-/- mice, whereas such increment of atherosclerosis was markedly attenuated after transplantation of bone marrow derived macrophages from ApoE-/-mice. Mechanistically, intracellular HMGB2 repressed IL-1 transcription and induced autophagic degradation of NLRP3, thus suppressing NLRP3 inflammasome priming and activation. Extracellular HMGB2 levels increased in progressive atherosclerotic plaques with macrophage apoptosis. Recombinant HMGB2 protein promoted inflammation in vitro via RAGE and intraperitoneal administration of this protein exacerbated atherosclerosis in mice.

Conclusion
This study indicates that decreased HMGB2 level in macrophage is associated with inflammation and atherosclerosis in patients with CAD. Intracellular HMGB2 inhibits NLRP3 inflammasome and atherosclerosis in mice, whereas released HMGB2 promotes inflammation and atherosclerosis in mice.
Parathyroid hormone exacerbates pulmonary hypertension via parathyroid hormone receptors on pulmonary artery smooth muscle cells
Authors: Joki Y , Konishi H , Takasu K , Katsuumi G , Yoshida Y , Furihata T , Minamino T 
Published: October 2024

Abstract

Background
Pulmonary hypertension (PH) is a right heart failure disease and can be fatal. Parathyroid hormone (PTH) is a bone metabolism hormone and regulates calcium and phosphorus homeostasis. Previous studies have suggested that PTH affects the cardiovascular system and influences cardiovascular diseases. We hypothesized that PTH may play a role in the pathogenesis of PH.

Purpose
This study aimed to investigate whether PTH has a critical role in pulmonary hemodynamics.

Method
Serum PTH levels were measured in patients with PH or suspected PH undergoing right heart catheterization. We tested whether the regulation of PTH and the PTH receptor (PTH1R) affected PH in hypoxia (Hx)-induced PH model mice and Sugen/hypoxia (SuHx)-induced PH model ras. Hx mice were treated with PTH daily for 3 weeks or with an inhaled AAV-shRNA system to knockdown PTH1R expression in the lung. SuHx rats underwent parathyroidectomy (PTx). We measured physical data and right ventricular systolic pressure (RVSP) in these models. Human pulmonary artery smooth muscle cells (PASMCs) were cultured and treated with PTH to examine the direct effects of PTH.

Result
In the clinical study, we enrolled 30 participants and found serum PTH levels correlated with mean pulmonary artery pressure (r = 0.67) and pulmonary vascular resistance (r = 0.62). The ROC curve showed a cut-off PTH level of 46.0 pg/mL (68.2% sensitivity, 100% specificity) to predict PH. In animal models, PTH treatment exacerbated right ventricular hypertrophy and increased right ventricular systolic pressure (RVSP) in Hx mice compared with non-treated Hx mice (Figure 1). In contrast, knockdown of PTH1R in the lungs improved PH in Hx mice, and PTx significantly suppressed PH in SuHx rats (Figure 2). PTH promoted migration and proliferation through PTH receptor-ERK signaling in PASMCs.

Conclusion
Our clinical and experimental data showed that PTH exacerbates PH and suggest that PTH/PTH1R signaling would be a new target for PH treatment.
Suppression of Fibulin 5 Attenuated Pulmonary Arterial Hypertension Associated with Congenital Heart Disease via Mitigating Pulmonary Arterial Smooth Muscle Cell Dysfunction
Authors: Yang Y , Shu S R , Wang P Z , Zhou J J 
Published: October 2024

Abstract
Backgrounds: The specific molecular pathogenesis of pulmonary arterial hypertension associated with congenital heart disease (CHD-PAH) and the reversal still remain elusive. Fibulin 5 was involved in the regulation of cardiovascular system while little is known about the expression and functions of fibulin 5 in the development of CHD-PAH. This study aimed to investigate the role of fibulin 5 in the pathogenesis of the disease.

Methods
Lung samples were obtained both from patients with CHD-PAH and aortocaval shunt-associated PAH rat models. TGF-β1was used to induce human pulmonary artery smooth muscle cells (hPASMCs) dysfunction and lentiviruses were responsible for alteration of fibulin 5 expression. In an effort to explore the specific role of fibulin 5, an inhibitor of phosphatidylinositol-3-kinases (PI3K) /protein-serine-threonine kinase (AKT) which was defined as the TGF-β1-related downstream signaling was applied for further exploration.

Results
Fibulin 5 was pronouncedly elevated in the PASMCs of the remodeled pulmonary arterioles from patients with CHD-PAH and shunt-associated PAH rats. We found that over-expression of fibulin 5 could promote hPASMCs dysfunction induced by TGF-β1 and it could be reversed by the PI3K/AKT inhibitor LY294002 suggesting the essential role of TGF-β1/PI3K/AKT signaling activation in phenotypic switch, proliferation and migration of PASMCs. Moreover, an adeno-associated virus vector encoding fibulin 5 by intratracheally instillation with rats improve the hemodynamic parameters in shunt-associated PAH rats and its related pulmonary artery remodeling.

Conclusion
Over-expression of fibulin 5 could significantly promote hPASMCs dysfunction and pulmonary artery remodeling via reinforcing TGF-β1/PI3K/AKT signaling activation, which highlighted the potential valuable pharmacological target for the treatment of CHD-PAH.Fibulin 5 was over-expressed in PAHInhibition of fibulin 5 attenuated PAH