bluuebunny/crossref_metadata_2025_split huggingface dataset:
Long COVID-19 syndrome: association of cardiopulmonary impairment with a persistent platelet activation
Authors: Camera M , Brambilla M , Canzano P , Becchetti A , Conti M , Agostoni P G , Pengo M , Tortorici E , Mancini M E , Andreini D , Bonomi A , Parati G F 
Published: October 2022

Abstract

Background
A considerable proportion of patients do not fully recover from COVID-19 infection and report symptoms that persist beyond the initial phase of infection: this condition is defined long-COVID-19 syndrome (LCS). LCS can involve lungs as well as several extrapulmonary organs, including the cardiovascular system. The risk and 1-year burden of cardiovascular diseases (CVD) is increased in COVID-19 survivors, even in subjects at low risk of CVD. Recently, we documented that acute COVID-19 infection induces altered platelet activation state characterized by a prothrombotic phenotype and by the formation of platelet-leukocyte aggregates (PLA), that may be involved in the pulmonary microthrombi found in autoptic specimens. No data are yet available on the contribution of platelet activation to residual pulmonary impairment and procoagulant potential in LCS patients.

Purpose
To study platelet activation status, microvesicle (MV) profile, platelet thrombin generation capacity (pTGC) in LCS patients enrolled at 6 months after resolution of the acute phase (6mo-FU), compared to acute COVID-19 infection patients.

Methods
6mo-FU COVID-19 patients (n=24) with established LCS were enrolled at Centro Cardiologico Monzino. Residual pulmonary impairment was assessed by Cardiopulmonary Exercise Testing (CPET) and 64-rows-CT scan evaluation. Platelet activation (P-selectin, Tissue Factor [TF] and PLA) and MV profile were assessed by flow cytometry; pTGC by calibrated automated thrombogram. 46 patients enrolled during acute COVID-19 infection and 46 healthy subjects (HS) were used for comparison.

Results
Dispnea in LCS patients was confirmed by CPET showing compromised alveolus-capillary membrane diffusion and residual pulmonary impairment. TF+-platelet and -MV levels were 3-fold (1.5% [1.2–2.9] vs 2.4% [1.6–5.7]) and 2-fold (217/μl [137–275] vs 435/μl [275–633]) lower at 6mo-FU compared to acute phase, being comparable to HS. pTGC behaved similarly. At 6mo-FU, the MV profile, in terms of total number and cell origin, returned to physiological levels. Conversely, although lower than that measured in acute phase, a 2.5-fold higher platelet P-selectin expression (6.9% [3–13.5] vs 11.7% [5.2–18.9]) and PLA formation (35.5% [27.4–46.8] vs 67.7% [45.7–85.3]) was observed at 6mo-FU compared to HS. Interestingly, a significant correlation between PLA formation and residual pulmonary impairment was observed (r=−0.423; p=0.02).

Conclusion
These data strengthen the hypothesis that the presence of PLA in the bloodstream, and thus also in the pulmonary microcirculation, may contribute to support pulmonary dysfunction still observed in LCS patients.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Italian Ministry of Health (Ricerca Corrente 2020 MPP COVID4)
Investigation of the effect of the angiotensin II type 2 receptor (AT2R) agonist C21 on plasma NT-proBNP, a diagnostic biomarker, in subjects hospitalised with COVID-19
Authors: Batta R , Tornling G , Rosendahl E , Bengtsson T , Raud J 
Published: October 2022

Abstract

The prohormone N-terminal pro-B-type natriuretic peptide (NT-proBNP) is released from stretched cardiac myocytes and is a diagnostic biomarker for heart failure and cardiac dysfunction as well as pulmonary embolism and pneumonia that are frequent complications to severe Coronavirus Disease 2019 (COVID-19). NT-proBNP is frequently elevated in COVID-19. In a recent publication, it was demonstrated that NT-proBNP was strongly associated with mortality in patients with COVID-19, and further investigation of its usefulness as a prognostic tool to predict disease outcomes in COVID-19 was suggested (1).
In the recently completed phase 2 trial (angiotensin II type 2 receptor agonist COVID-19 trial [ATTRACT]; NCT04452435) in subjects hospitalised with COVID-19, it was investigated whether treatment with the AT2R agonist C21 for 7 days affected the release of the plasma biomarker NT-proBNP.
ATTRACT was a randomised, double-blind, placebo-controlled, phase 2 trial that investigated the safety and efficacy of C21 treatment (100 mg twice daily) for 7 days in hospitalised subjects with COVID-19, not requiring mechanical ventilation. The results of the trial demonstrated that treatment with C21 on top of standard of care (vast majority of patients received glucocorticoids) significantly reduced the proportion of subjects requiring supplemental oxygen at Day 14, indicating faster recovery with C21 treatment compared to placebo. Blood samples for exploratory analysis were taken before and after 7 days of treatment with C21 or placebo.
Plasma NT-proBNP was markedly elevated in both treatment groups before treatment, with average values of 357 and 438 pg/mL in the placebo and C21 groups, respectively, as compared to normal levels of approximately &lt;100 pg/mL. After 7 days of treatment, the C21 group experienced a dramatic reduction in plasma NT-proBNP (by 259 pg/mL) as compared to the placebo group (63 pg/mL) (p=0.02).
The results show that short-term C21 treatment decreased the release of NT-proBNP in subjects hospitalised with COVID-19. Further investigations are needed to elucidate whether this is related to effects on COVID-19-induced pulmonary damage or direct protective effects on the heart. We are currently conducting a global phase 3 trial (VP-C21–008) further investigating the effect of C21 in subjects hospitalised with COVID-19 including determination of NT-proBNP.

Funding Acknowledgement
Type of funding sources: Private company. Main funding source(s): Vicore Pharma AB
Identification of a PRDM1-regulated T cell network to regulate T cell driving plaque inflammation in human and mouse atherosclerosis
Authors: Maas S L , Jin H , Lu C , Nagenborg J , Karel J M H , Cavill R , Sikkink C J J M , Nadeau S , Gijbels M , Mees B M E , Smirnov E , Sluimer J C , Martins G A , Van Der Vorst E P C , Biessen E A L 
Published: October 2022

Abstract

T cells have a prominent role in the pathogenesis of atherosclerosis, although their exact function remains elusive. Here, we pursued a network-driven approach to identify T cell-associated gene programs driving the transition from low- to high-risk human plaques.
In this study 43 human carotid arterial plaques were collected and stratified based on absence (low-risk) or presence (high-risk) of intraplaque haemorrhage (IPH). Lesion RNA was subjected to microarray gene expression analysis and analysed by Weighted Gene Co-expression Network Analysis (WGCNA). We identified a co-expressed gene cluster displaying a strong T cell signalling signature in high- versus low-risk plaque, which was tightly connected to subnetworks of angiogenesis and interferon-signalling. WGCNA-based Bayesian network inference, cell-type deconvolution and single-cell gene expression revealed that this T cell-associated gene program was likely linked to effector-memory cytotoxic CD8+ T cells, underpinning the central role of T cells in plaque destabilization. Gene regulatory analysis identified cytotoxic T cell-related transcription factors, like PRDM1, regulating this plaque T cell gene program. Moreover, we demonstrated in LDL receptor knockout mice with T cell-specific Prdm1 deficiency, that lack of Prdm1 in T cells resulted in larger, more advanced plaques.
In conclusion, our study reveals a PRDM1-regulated T cell footprint in high- versus low-risk human atherosclerotic lesions and murine atherosclerotic plaque development, thereby identifying this network as a potential target for intervention in adverse T cell responses.

Funding Acknowledgement
Type of funding sources: Public grant(s) – EU funding. Main funding source(s): The European Research Area Network on Cardiovascular Diseases (ERA-CVD and Dutch Heart Foundation)
Role of smooth muscle-derived S100A4 in an atherosclerotic mouse model
Authors: Cardoso Dos Santos L M , Azar P , Lemeille S , Bochaton-Piallat M L 
Published: October 2022

Abstract

Background
S100A4, a small calcium binding protein, plays an important role in vascular smooth muscle cell (SMC) phenotypic switch but its implication in atherosclerotic plaque development and particularly in SMC phenotypic plasticity is not clear yet. By neutralizing S100A4 using a monoclonal antibody, we have previously shown a reduction in overall atherosclerotic burden. However, this strategy did not distinguish the different contribution between SMCs or other S100A4-expressing cells (e.g. macrophages).

Methods
Herein, we used a lineage tracing mouse model in which we induced a SMC specific deletion of S100A4 (SMC-S100A4Δ/Δ) in an ApoE−/− background. High cholesterol diet was maintained for 12 weeks, after which, SMC-S100A4Δ/Δ and control mice (SMC-S100A4wt/wt) were sacrificed and aortas processed for staining and single cell RNA sequencing (scRNA-seq).

Results
We showed that S100A4 deletion modulated plaque composition rather than plaque size. ScRNA-seq analysis from SMC-S100A4Δ/Δ and SMC-S100A4wt/wt showed a reduction in macrophages and dendritic cells and an increase in total SMCs, likely participating in fibrous cap stabilization. SMC populations showed a decrease in macrophage-like inflammatory phenotype and an increase in fibroblast-like repair phenotype, as well as retention of specific SMC markers, namely Acta2 and Myh11. Gene expression analysis revealed a reduction in the expression of inflammation markers, fatty acid metabolism, and extracellular matrix-related genes, and an increase in classical SMC markers and maintenance of the contractile phenotype.

Conclusion
We report an intrinsic function of S100A4 in the establishment of SMC fate within the plaque and, surprisingly, a global impact on the atherosclerotic plaque microenvironnement and inflammation status.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Swiss National Science Foundation
Targeting RNA elongation in endothelial-to-mesenchymal transition
Authors: Hoba J , Kokot K E , Andritschke M , Boeckel J N , Laufs U 
Published: October 2022

Abstract

Background and purpose
EndMT describes a process in which endothelial cells lose their endothelial features such as the maintenance of the vascular barrier function and trans-differentiate into mesenchymal like cells. ECs which have underwent EndMT are enriched in vulnerable plaques in humans.
Several stress stimuli such as hypoxia or inflammation demand immediate induction of adaptive gene expression to improve cell survival. In contrast to the usual induction of gene expression which requires several hours, release of paused RNA Polymerase II (RNAPII) from promoter allows rapid gene induction via RNA elongation. This rapid response mechanism is mediated by the super elongation complex (SEC), which is built around the scaffold proteins AFF1 and AFF4.
Here, we hypothesized that RNA elongation regulates EndMT and therefore investigated on the contribution of the SEC scaffold proteins during the process of EndMT.

Methods and results
After induction of EndMT in human primary endothelial cells (EC) the elongation factors AFF1 and AFF4 were induced (AFF1: 1.9-fold and AFF4: 2.7-fold, P&lt;0.05). Overexpression of AFF4 resulted in increased expression of the mesenchymal marker genes SM22 (1.8-fold, P&lt;0.05) and VCAN (2.0-fold, P&lt;0.05). Accordingly, treatment of EC with two small molecules inhibiting the SEC reduced the expression of several mesenchymal markers SM22 (−87.6%, P&lt;0.05) and Calponin (−93.8%, P&lt;0.05) during EndMT. After induction of EndMT, a large number of newly synthesized RNA transcripts were upregulated, indicating regulation of gene expression on the level of RNA elongation during EndMT. Sequencing of newly transcribed RNA (4sU-sequencing) after treatment with the SEC inhibitors revealed a lower transcript density downstream of promoter sites, suggesting a decreased RNA elongation rate in ECs. SEC inhibitors decreased the expression of the majority of EndMT-induced transcripts (63.6%). Among these rapid induced genes we found the mRNA of RE1 (+10,8-fold). We validated the regulation of RE1 by qPCR (2.8-fold, P&lt;0.05) and single-cell sequencing. Chromatin immunoprecipitation (ChIP) sequencing revealed a reduced occupation of RNAPII in proximity of promotor sites of RE1 upon induction of EndMT. These data suggest a SEC-mediated regulation on the level of RNA elongation of RE1 during EndMT.

Conclusion
SEC scaffold proteins AFF1 and AFF4 are upregulated during EndMT. Targeting the SEC during EndMT reduces the expression of common mesenchymal markers, consistent with an overall higher occupancy of paused RNAPII at promoter-proximal sites. We identify RE1 as a rapidly induced gene during EndMT that is mediated by the SEC scaffold proteins AFF1 and AFF4. Our findings suggest that RNA elongation is an inducer of EndMT.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)
Arterial thrombosis in Hutchinson-Gilford Progeria Syndrome
Authors: Puspitasari Y M , Ministrini S , Karch C , Akhmedov A , Luscher T F , Camici G G 
Published: October 2022

Abstract

Introduction
Arterial thrombosis is the most common age-associated event underlying major adverse cardiovascular (CV) events. The interplay between the vascular endothelium, platelets, and the coagulation cascade leads to thrombus formation, which results in the cessation of blood supply to the downstream tissues. Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic condition with striking features of premature aging. It is caused by defects in the nuclear A-type lamin gene, leading to intracellular accumulation of progerin. This genetic disorder is characterized by shortened lifespan, primarily due to an increased incidence of myocardial infarction and ischemic stroke. Declined vascular function and compliance have been reported in HGPS patients. Nevertheless, the effect of the specific A-type lamin gene mutation on coagulation and thrombus formation has not been investigated previously.

Methods
28- to 30-week-old male and female transgenic heterozygous LmnaG609G knock-in (HGPS) mice and corresponding wild-type (WT) littermate controls were exposed to photochemically-induced carotid artery endothelial injury to trigger arterial thrombosis. Vascular and circulating levels of tissue factor (TF), plasminogen activator inhibitor (PAI)-1, and von Willebrand factor (vWF) were measured using enzyme-linked immunosorbent assay (ELISA). TF activity assay was also performed on carotid artery homogenates of WT and HGPS animals.

Results
HGPS mice displayed accelerated thrombus formation compared to the WT animals as underlined by a shortened time to occlusion. Although this finding suggests an increased activation of the extrinsic coagulation cascade, no significant differences were found in TF expression and activity in carotid artery lysates. Circulating and vascular expression of the fibrinolytic factor PAI-1 was also found to be similar between WT and HGPS animals. Furthermore, no significant difference in plasma vWF between the two groups was observed.

Conclusions
Our results show an increased arterial thrombotic response in HGPS mice as compared to WT littermates. This novel observation could provide a mechanistic explanation for the increased incidence of acute cardiovascular events observed in HGPS patients. Further studies will be conducted to investigate the molecular mechanism underlying the observed effects, in particular, on the potential involvement of platelets.

Funding Acknowledgement
Type of funding sources: None.
Totum-070 prevents diet-induced hypercholesterolemia in western diet fed mice and inhibits cholesterol uptake in human enterocyte caco2 cell line
Authors: Langhi C , Otero Y F , Le Joubioux F , Guigas B , Peltier S , Sirvent P 
Published: October 2022

Abstract

Introduction
It is well known that hypercholesterolemia is an important modifiable risk factor in the development of cardiovascular diseases. Lowering blood cholesterol level reduces the incidence of atherosclerotic disease. Totum-070 (T070) is a clinical stage polyphenol-rich active substance composed by the association of 5 plant extracts.

Purpose
We assessed the hypothesis that administration of T070 prevents hypercholesterolemia in western diet fed mice and explored the potential mechanisms involved in vitro in human enterocytes.

Methods
C57BL/6 mice were fed either a normal diet (ND) or western diet (WD) for 6 weeks and received either vehicle or T070 (1mg/kg or 3mg/kg) daily by gavage (n=14 mice per group). Food intake, body weight and plasma lipid were monitored along study duration. Liver and feces lipid content were quantified at the end of the study. For in vitro studies, human Caco2 cells were differentiated in transwells for 21 days. To perform cholesterol uptake assay, [1,2-3H(N)]-cholesterol was incubated in apical medium for 1 hour in presence or not of T070 prior to quantification of radioactivity in the cells.

Results
In mice, supplementation with T070 had no effect on food intake during the study. At the end of the in vivo study, body weight in WD-fed mice was increased to 28.3±0.8 g compared to the ND group (24.5±0.4 g, p&lt;0.001). Interestingly, a significant (p&lt;0.01) 29% reduction in body weight gain was observed in the T070 3mg/kg group compared to the WD group at the end of the study. Compared to WD group, total-cholesterol in mice supplemented by T070 was reduced in a dose dependent manner (298±8 mg/dl WD vs 276±9 mg/dl T070 1mg/kg, p=0.19, and 265±8 mg/dl T070 3mg/kg, p&lt;0.05). Furthermore, hepatic steatosis induced by the WD was reduced by 73% (p&lt;0.001) in the T070 3mg/kg group. Hepatic and intestinal gene expression were drastically altered by WD feeding compared to ND, while supplementation with T070 tended to restore normal expression levels for many genes implicated in lipid and lipoprotein metabolism. A significant increase in fecal cholesterol excretion was observed in mice supplemented with T070 3 mg/g compared to the mice fed with WD alone (442.3±151.7 nmol/day WD + T070 3mg/g vs 75.4±5.9 nmol/day WD, p&lt;0.05). Studies in human enterocytes demonstrated that 1 g/l T070 reduced cholesterol uptake by 35% (p&lt;0.001), suggesting that T070 inhibits intestinal cholesterol absorption.

Conclusion
Our results show the beneficial effect of supplementation with Totum-070 to prevent induction of hypercholesterolemia in a nutritional mouse model. Investigation of mechanisms of action in enterocytes demonstrate the role of Totum-070 to inhibit intestinal cholesterol absorption. These results highlight the interest of using Totum-070 for the management of hypercholesterolemia.

Funding Acknowledgement
Type of funding sources: None.
Ceramide-based lipid profiles and the prevalence of type 2 diabetes differ between patients with coronary artery disease and those with peripheral artery disease
Authors: Leiherer A , Muendlein A , Brandtner E , Saely C H , Vonbank A , Mader A , Sprenger L , Maechler M , Jylha A , Laaperi M , Laaksonen R , Maerz W , Fraunberger P , Kleber M , Drexel H 
Published: October 2022

Abstract

Introduction
Serum lipids and metabolic diseases, in particular type 2 diabetes (T2D) and non-alcoholic fatty liver disease (NAFLD), predict the atherosclerotic diseases coronary artery disease (CAD) and peripheral arterial disease (PAD).

Purpose
The purpose of this study was to investigate in how far a more detailed characterization including serum lipids improves discrimination of PAD from CAD.

Method
A cohort of 274 statin-naïve patients with either PAD (n=89) or stable CAD (n=185) were referred to metabolic screening and were characterized using nuclear magnetic resonance- and liquid chromatography-tandem mass spectrometry based advanced lipid and lipoprotein analysis. Results were validated in an independent cohort of 1239 patients with PAD or CAD.

Results
We found a significant difference in T2D prevalence and in the ceramide-based lipid profile between PAD and CAD patients. However, neither cholesterol-based markers (including LDL-C, HDL-C) and detailed lipoprotein profiles nor the LD status differed significantly between PAD and CAD patients (figure). The difference between ceramide-based lipid profiles of CAD and PAD remained significant also after adjusting for body composition, smoking, inflammatory parameters, and T2D.

Conclusion
We conclude that PAD and CAD differ in ceramide-based lipid profiles and T2D status, but not in other lipid characteristics or metabolic diseases.

Funding Acknowledgement
Type of funding sources: None.
Acute arterial haemodynamics activation of endothelial to mesenchymal transition in long saphenous veins. Impact on vein graft disease
Authors: Ladak S , McQueen L , Joeldavid L , Murphy G , Zakkar M 
Published: October 2022

Abstract

Introduction
The long saphenous vein (LSV) is frequently used in cardiac surgery; however, its use is complicated by late stenosis or occlusion due to the development of intimal hyperplasia (IH). TGF-β has been implicated in the process of IH however the impact of acute haemodynamic changes on the activation of TGF-β endothelial-to mesenchymal transition (EndMT) has not been assessed and it's the focus of this study.

Purpose
To assess the role of acute haemodynamics changes in veins implanted into arterial circulation on EndMT.

Methods
Surplus LSV were exposed to acute arterial haemodynamics using a perfusion bioreactor. Changes in EndMT markers at the RNA and protein level evaluated by quantitative real time PCR, RNAScope and immunofluorescence.

Results
The acute exposure of veins to arterial haemodynamics ex-vivo induced significant increase in inflammatory marker IL-8 expression and transcription factor TWIST1 in LSV (both p≤0.01, Figure 1A) endothelium. Furthermore, immunostaining demonstrated the activation of pSMAD (p≤0.01, Figure 1B) acutely in endothelium after 45 minutes of exposure to arterial haemodynamics. This was followed by significant increase of SMC related markers (Vimentin and α-SMA; Figure 1C, both p≤0.001) and the suppression of endothelial cell related marker CD31 after 4 hours of LSV exposure to acute arterial haemodynamics. RNAScope and IHC results showed localisation of TWIST1 RNA and protein in CD31+ (p≤0.001) and VECAD+ (p≤0.001) cells respectively following exposure to acute arterial haemodynamics (Figure 1D). Furthermore, TGF-β pathway phosphorylation array identified the activation of Smad1 (p≤0.05) but not TAK1 in LSV endothelial cells indicating that acute arterial haemodynamics activates the TGF-β–SMAD pathway specifically (Figure 1E).

Conclusion
The exposure of LSV to acute arterial haemodynamics is associated with the activation of TGF-β–SMAD pathway leading to EndMT changes in the endothelium of vein grafts ex-vivo. This contributes to our understanding of the changes that occur in veins after implantation into arterial circulation and that the acute changes in the endothelium and suggests that strategies to modulate TGF-β–SMAD can be utilised to modulate IH in vein grafts.

Funding Acknowledgement
Type of funding sources: Private grant(s) and/or Sponsorship. Main funding source(s): Van Geest Foundation Heart and Cardiovascular Diseases Research Fund