bluuebunny/crossref_metadata_2025_split huggingface dataset:
Long non-coding RNA TERRA regulates DNA damage and survival of endothelial cells and cardiomyocytes: implications for aging
Authors: Bink D I , Pham T P , Van Bergen A , Stanicek L , Van Der Ven D , Groenen E M J , Kaagman L , Jonk M , Hofmann P , Dimmeler S , Boon R A 
Published: October 2022

Abstract

Introduction
Ageing is the major risk factor for cardiovascular disease. Long non-coding RNAs are emerging as novel regulators of cellular functions and contributors to cardiovascular ageing. One of the hallmarks of aging is telomere attrition. Non-coding transcripts called Telomeric repeat-containing RNA (TERRA) are molecules of 0.2–10kb in length which are transcribed from the subtelomeres and telomeres of chromosomes and might play a role in cardiovascular ageing.

Purpose
This study aims to characterize the role of TERRA in aging of the cardiovascular system.

Methods and results
TERRA molecules from different chromosomes were upregulated in the hearts of old mice compared to young mice (p=0.002). Increased TERRA expression was also shown in heart tissue of patients with ischemic heart disease compared to donors (p=0.001). In vitro an upregulation of the TERRA molecule transcribed from chromosome 20 (h20q-TERRA) was found with increasing passage in human umbilical vein endothelial cells (HUVECs) (p=0.014) and IPSC-derived cardiomyocytes (p=0.011). After h20q-TERRA knockdown with LNA GapmeRs, HUVECs show less sprout formation in a spheroid assay compared to negative control transfected HUVECs (p=0.002), without showing a change in migration (p=0.205) or proliferation (p=0.114). H20q-TERRA knockdown revealed an increase in apoptosis (p=0.015) and telomeric DNA damage (p=0.011) and a decrease in telomere length (p&lt;0.001), while lentiviral TERRA-repeat overexpression had the opposite effect (p=0.016, p=0.031, p&lt;0.001, resp.). Apoptosis (p=0.012) and telomeric DNA damage (p=0.007) were also increased after the knockdown of h20q-TERRA in human cardiomyocytes. An apoptosis pathway profiler array in HUVECs showed that the expression of the antioxidant PON2 was decreased after knockdown of h20q-TERRA (p=0.040). PON2 expression was increased after TERRA overexpression (p=0.003). RNA immunoprecipitation revealed that TERRA can bind to PON2. Silencing PON2 in TERRA overexpressing cells diminished the TERRA-mediated decrease in caspase activation, suggesting a detrimental role for PON2 in caspase activation and endothelial cell survival.

Conclusion
Our data demonstrates that TERRA is upregulated with ageing and plays a role in endothelial and cardiomyocyte function and survival.

Funding Acknowledgement
Type of funding sources: Public grant(s) – EU funding. Main funding source(s): Horizon 2020
Genome-wide association study on cerebral white-matter hyperintensities in 36,577 individuals
Authors: Jespersen J , Ahlberg G , Andreasen L , Ghouse J , Frederiksen K S , Haunsoe S , Svendsen J H , Frikke-Schmidt R , Olesen M S , Bundgaard H 
Published: October 2022

Abstract

Background/Introduction
A quarter of all strokes are caused by cerebral small vessel disease (CSVD), which is also the most common pathology underlying vascular dementia. [1] White matter hyperintensities (WMH), a radiological marker detectable on magnetic resonance imaging (MRI), can serve as a proxy for CSVD. WMH is associated with stroke, increased risk of dementia and functional decline in older age. [2,3]

Purpose
We aimed at investigating the genetic architecture of WMH using cerebral MRI data.

Methods
We used imaging data ∼40,000 individuals from the UK Biobank, a large population-based, prospective cohort study. We selected individuals with available total volume of WMH from T1 and T2_FLAIR images.
We performed a genome-wide association study (GWAS) on autosomal genetic variants assuming an additive model based on genotype dosages with BOLT-LMM treating WMH as outcome.
We applied LD score regression (LDSC software) to estimate the genetic correlation between WMH traits and traits selected based on availability and relevance for cardiovascular disease.

Results
We included a total of 36,577 individuals with available quality controlled cerebral MRI data.
Genome-wide analysis identified 20 loci of statistical significance, six of which are not previously reported (Fig. 1). Within these novel loci, the following genes are located in proximity to lead variants: EHBP1, OTX1, WDPCP, VCAN, WNT16, FAM3C, ERI1, PRAG1, CACNB2, MTHFSD, FOXL1, FOXC2. Using BOLT, we found the heritability of WMH to be 37%.
We investigated genetic correlation between WMH and multiple phenotypes (Fig. 2). We observed the highest correlation with small vessel stroke (rg=0.56, P=3.9x10–4) Interestingly, WMH was genetically correlated with left atrial volume (rg=0.24, P=2.8x10–3).

Conclusion
In a GWAS on WMH acquired from cerebral MRI, we identified 20 significant loci, of which six are novel. This genetic study provides insights on the biological understanding and epidemiology of CSVD.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): John and Birthe Meyer FoundationThe Hallas-Møller Emerging Investigator Novo Nordisk
Adverse epigenetic signatures driven by histone methyltransferase EZH2 contribute to hyperglycaemia-induced endothelial dysfunction
Authors: Sanchez-Ceinos J , Hussain S , Waheed-Khan A , Zhang L , Pernow J , Cosentino F 
Published: October 2022

Abstract

Introduction
Posttranslational modifications of histones play a critical role in regulating gene expression in health and disease. Activation of histone methyltransferase enhancer of zeste homolog 2 (EZH2) catalyses repressive trimethylation of histone 3 at lysine residue 27 (H3K27me3) and favours reactive oxygen species (ROS) generation as well as pro-inflammatory changes. Oxidative stress and inflammation are linked to hyperglycaemia-induced endothelial dysfunction.

Purpose
The present study was designed to investigate whether EZH2 may represent a novel therapeutic target against hyperglycaemia-induced abnormal phenotypes in the endothelium.

Methods
The effect of EZH2 was tested in aortas from male mice (3–5 months old), human aortic endothelial cells (HAECs) and aortic endothelial cells isolated from diabetic individuals (D-HAECs) in the presence and in the absence of pharmacological inhibitor GSK126 (5 μM) or gene silencing of EZH2. HAECs and aortic rings were exposed either to normal glucose (5 mM) or high glucose (25 mM) for 20 hrs. Expression levels of H3K27me3, EZH2, oxidant and pro-inflammatory genes were determined by RT-PCR, western blotting, and protein activation assay kits. Superoxide anion production was measured by electron spin resonance spectroscopy. Binding of H3K27me3 to the promoter region of candidate genes was evaluated by chromatin immunoprecipitation (ChIP). Endothelium-dependent and independent relaxations were assessed in aortic rings by organ chamber experiments.

Results
Immunoblotting showed a significant increase in EZH2 activity measured as H3K27me3 in HAECs exposed to high glucose and in D-HAEC. ChIP assays showed that binding of H3K27me3 to the promoter of anti-oxidant and transcription factor JunD genes triggers ROS generation and activation of inflammatory pathways. Interestingly enough, histone methyltransferase EZH2 inhibitor GSK126 or specific siRNA blunting H3K27me3 abolished hyperglycaemia-induced oxidative stress and inflammation. Moreover, targeting EZH2 rescued endothelial function.

Conclusions
These results uncover adverse epigenetic signatures underlying endothelial dysfunction in diabetes. Targeting EZH2 may attenuate oxidative and inflammatory transcriptional programmes and thus prevent vascular disease in this setting.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): Swedish Research Council, the Swedish Heart & Lung Foundation, and King Gustav V and Queen Victoria Foundation.
Fast and more affordable CYP2C19 LOF testing to personalized of clopidogrel therapy
Authors: Akkaif M A , Sha'aban A , Daud N A A , Noor D A M , Musa N F , Sk Abdul Kader M A , Ibrahim B 
Published: October 2022

Abstract

Background
Clopidogrel is an antiplatelet drug used to treat coronary artery disease (CAD) to reduce blood clotting by inhibiting platelet activity. However, many patients experience ischemic events due to changes in clopidogrel metabolism attributed to various genetic and non-genetic factors. CYP2C19 is one of the important genes responsible for the metabolism of clopidogrel, but this gene has polymorphisms in cytochrome P450 (CYP), and this CYP2C19 Loss-of-function (LoF) polymorphisms impair clopidogrel metabolism between individuals and races. Therefore, genetic testing is an effective strategy to personalized clopidogrel and reduce the incidence of ischemic events in CAD patients.

Purpose
We sought to develop a nested allele-specific multiplex polymerase chain reaction (PCR) method for the detection of CYP2C9 LOF alleles.

Method
Genomic DNA was extracted from blood taken from 7 healthy volunteers using a commercial DNA extraction kit. A two-step PCR method was developed. First, DNA was subjected to a first PCR used to amplify exons 4 and 5 simultaneously in a single reaction tube. Second, The products of 1st PCR were then used as a template in the second PCR to detect CYP2C9 polymorphisms alleles (CYP2C19*2 and *3) using allele-specific primers. The test results were validated via sequencing.

Result
We successfully detected clinically important CYP2C19 LOF polymorphisms alleles by the nested allele-specific multiplex PCR method (Figure 1). This method is fast and doesn't require high-quality and expensive laboratory equipment. It was reproducible and specific and can reliably detect CYP2C19 variants. Direct sequencing of target variants has been used to validate this strategy further, and the amplified sequences were 100% identical to the CYP2C9 sequence.

Conclusion
The developed methods were specific to identify polymorphisms in CYP2C19 (rs4244285 and rs4986893). However, to understand ethnic differences and their potential implications for clopidogrel therapy, we highly recommend identifying relevant genetic polymorphisms before clinical manifestations arise.

Funding Acknowledgement
Type of funding sources: None.
Circadian nuclear receptor Rev-erbalpha is expressed by platelets and potentiates platelet activation and thrombus formation
Authors: Shi J , Tong R , Zhou M , Gao Y , Zhao Y , Chen Y , Liu W , Li G , Lu D , Meng G , Hu L , Yuan A , Lu X , Pu J 
Published: October 2022

Abstract

Background
Adverse cardiovascular events have day/night patterns with peaks in the morning, potentially related to endogenous circadian clock control of platelet activation. Circadian nuclear receptor Rev-erbα is an essential and negative component of the circadian clock.

Purpose
We aim to investigate the expression profile and biological function of Rev-erbα in platelets.

Methods and results
Here we report the presence and functions of circadian nuclear receptor Rev-erbα in human and mouse platelets. Both human and mouse platelet Rev-erbα showed a circadian rhythm that positively correlated with platelet aggregation. Global Rev-erbα knockout and platelet-specific Rev-erbα knockout mice exhibited defective in hemostasis as assessed by prolonged tail-bleeding times. Rev-erbα deletion also reduced ferric chloride-induced carotid arterial occlusive thrombosis, prevented collagen/epinephrine-induced pulmonary thromboembolism, and protected against microvascular microthrombi obstruction and infarct expansion in an acute myocardial infarction model. In vitro thrombus formation assessed by CD41-labeled platelet fluorescence intensity was significantly reduced in Rev-erbα knockout mouse blood. Platelets from Rev-erbα knockout mice exhibited impaired agonist-induced aggregation responses, integrin αIIbβ3 activation and α-granule release. Consistently, pharmacological inhibition of Rev-erbα by specific antagonists decreased platelet activation markers in both mouse and human platelets. Mechanistically, mass spectrometry and co-immunoprecipitation analyses revealed that Rev-erbα potentiated platelet activation via oligophrenin-1-mediated RhoA/ERM (ezrin/radixin/moesin) pathway.

Conclusion
We provide the first evidence that circadian protein Rev-erbα is functionally expressed in platelets and potentiates platelet activation and thrombus formation. Rev-erbα may serve as a novel therapeutic target for managing thrombosis-based cardiovascular disease.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): This work was supported by grants from the National Science Fund for Distinguished Young Scholars (81625002), the National Natural Science Foundation of China (81930007).
Thrombin receptor protease-activated receptor 1 (PAR1) related cytotoxic CD8+ T cell activity is associated with atrial myopathy and pro-inflammatory immune response in early atrial fibrillation
Authors: Friebel J , Wegner M , Lammel S , Bloebaum L , Jakobs K , Puccini M , Landmesser U , Rauch U 
Published: October 2022

Abstract

Background and aims
Cytotoxic CD8+ T cells mediate myocardial damage in patients with virus induced myocarditis, plaque erosion during acute coronary syndrome, and promote adverse post-ischemic cardiac remodeling. There is emerging evidence to support a link between inflammation and atrial fibrillation (AF). Protease-activated receptor 1 (PAR1) is the thrombin receptor expressed on platelets, but it is also expressed in endothelial cells, vascular smooth muscle cells, cardiac fibroblasts, and CD8+ T cells. The role of CD8+ T cells and PAR1 in atrial myopathy and early AF have not been studied.

Methods
In 80 non-anticoagulated patients with a first documented episode of AF, and 20 control subjects without AF but with a comparable cardiovascular risk profile (e.g. SCORE2, CHA2DS2-VASc, p&gt;0.05), we studied PAR1-related cytotoxic T cell activity in the first hours (up to 24 h) of AF clinical manifestation. The Apolipoprotein E–knock out (ApoEko) mouse is a commonly used model of metabolic syndrome that develops atrial myopathy (structural correlate of AF). ApoEko male mice (Apoetm1Unc with a C57BL/6 genetic background) were fed for six weeks with a standard chew, western-type diet (high fat and high sucrose = WD) ± the specific PAR1 inhibitor vorapaxar.

Results
High levels of circulating, activated CD8+ T cells (CD8+CD57+ T cells with pro-inflammatory and high cytotoxic properties) were present in patients with early AF (flow cytometry). Stimulation of cytotoxic T cells, that expressed the thrombin receptor PAR1, that were isolated from patients with new onset of AF, released pro-inflammatory cytokines (e.g. IFN-γ, TNF-α). Enhanced cytotoxic activity of CD8+ T cells was suggested by elevated plasma levels of effector molecules (e.g. granulysin and granzyme).
In ApoEko mice fed a WD, atrial myopathy as the structural correlate for AF, was paralleled by increased expression of effector molecules that were associated with CD8+ T cell activity (like perforin, granzyme A, granzyme B, IFN-γ). Cytotoxic T cell activity disrupted the release of the potective atrial natriuretic peptide, ANP, and increased signaling pathways associated with atrial fibrosis (e.g. TGF-β, cTGF, TGF-β receptor). The specific PAR1 inhibitor, vorapaxar, reduced inflammatory T cell activity, transcription of fibrotic mediators, and reconstituted ANP secretion in atria of ApoEko mice.

Conclusions
Targeting the FXa/FIIa-PAR1-CD8+ axis might be a promising approach to reduce atrial fibrosis and inflammation.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): BIH Charité Clinician Scientist Program
SWATH proteomic analysis from the standard care versus celecoxib outcome trial identifies renal biomarkers are associated with early cardiovascular events amongst NSAID users
Authors: Vaja R , Ferreira P M , Want L , Ahmetaj-Shala B , Gashaw H , Kirkby N S , Mitchell J A 
Published: October 2022

Abstract

Background
Despite being one of the most commonly used and readily available drugs worldwide, we still do not understand why non-steroidal anti-inflammatory drugs (NSAIDs) increase cardiovascular risk. A proteomics approach may provide mechanistic insight into how COX-2 protects the cardiovascular system.

Purpose
To use SWATH proteomics to identify cardiovascular biomarkers in NSAID users who go onto have a cardiovascular event.

Methods
A case control study was designed using the Standard Care Versus Celecoxib Outcome Trial (SCOT) biobank (1). Serum samples from 228 NSAID users with osteoarthritis were analysed. From these, an exploratory analysis of 15 individuals who had a cardiovascular event within 6 months were compared to 213 individuals who did not. SWATH proteomics was used to identify biomarkers and findings were validated by immunoassay.

Results
Seventy-eight proteins were detected at significantly different levels in serum from participants who did or did not have an event within 6 months of enrolment. Of these the greatest increase in the event group was in levels of the renal dysfunction marker, cystatin C (Log Fold difference 1.4, p&lt;0.001). Validation using a renal biomarker immunoassay panel confirmed a higher level of cystatin C (Z-score 0.6 vs −0.04, p: 0.02) as well as a lower level of uromodulin (Z-score −0.5 vs 0.03, p: 0.036) in the event group. In agreement, mass spectrometry analysis identified a higher level of the cardio-renal biomarker asymmetric dimethylarginine (ADMA) in the event group (Z-score 0.4 vs −0.03, p: 0.07) (Figure 1).

Conclusion
Renal biomarkers are associated with early cardiovascular events amongst NSAID users in the SCOT cohort. Thus, changes in renal function should be investigated as a mechanistic or predictive factor in this population at elevated cardiovascular risk.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): British Heart Foundation
Protective effects of COVID-19 vaccination on endothelial function of patients with COVID-19
Authors: Lampsas S , Oikonomou E , Souvaliotis N , Papamikroulis G A , Lytra T , Sarantos S , Mystakidi V C , Pantelidis P , Kalogeras K , Theofilis P , Tsatsaragkou A , Siasos G , Tsioufis C , Tousoulis D , Vavuranakis M 
Published: October 2022

Abstract

Background
Endotheliilitis COVID-19 related endothelial dysfunction plays a key role in the cardiovascular complications of the disease. Vaccine against SARS-CoV-2 protects against severe COVID-19 and from adverse effects. We evaluated the impact of vaccination on COVID-19 induced endothelial dysfunction.

Methods
We enrolled 45 patients hospitalized for COVID-19 (either vaccinated or not against SARS-CoV-2). Clinical information and laboratory findings were collected, and brachial artery flow-mediated dilation (FMD) was evaluated as a measure of endothelial function. Subjects without COVID-19 were used as the control group. All patients were hospitalized in a medical ward classified according to the World Health Organization (WHO) scale.

Results
There was no difference in age (62±10 years vs. 65±8 years, p=0.12) and male sex prevalence (56% vs. 49%, p=0.53) between patients with COVID-19 and control subjects. Of the patients with COVID-19, 44% (20) were vaccinated against SARS-CoV-2. FMD was impaired in patients with COVID-19 compared to controls (4.35±3.56% vs. 7.36±2.91%, p&lt;0.001). In patients with COVID-19, FMD was impaired in non-vaccinated subjects compared to vaccinated (2.05±2.41% vs. 7.24±2.52%, p&lt;0.001). There was no difference in FMD between controls and vaccinated against COVID-19 patients (7.36±2.91% vs. 7.24±2.52%, p=0.86). There was no difference in the WHO scale clinical status for vaccinated and not vaccinated COVID-19 subjects (For Vaccinated WHO scale 3: 35%; scale 4: 35%; scale 5: 30% vs. For Non-vaccinated WHO scale 3: 20; scale 4: 60%; scale 5: 20%, p=0.24).

Conclusion
Hospitalized patients with COVID-19 present endothelial dysfunction in the acute phase of the disease. Endothelial function in unvaccinated patients with COVID-19 is impaired compared to control subjects as well as compared to vaccinated patients with COVID-19. This data provides insights on the protective role of vaccination against COVID-19 related endotheliitis.

Funding Acknowledgement
Type of funding sources: None.
GPR55 deficiency in B-cells promotes atherosclerosis and regulates plasma cell maturation
Authors: Guillamat-Prats R , Hering D , Rami M , Haerdtner C , Santovito D , Rinne P , Pagano S , Nicolas Vuilleumier N , Schmid S , Janjic A , Enard W , Weber C , Maegdefessel L , Hilgendorf I , Steffens S 
Published: October 2022

Abstract

Atherosclerosis is a chronic and multifactorial disease accompanied by an imbalance between resolving and pro-inflammatory lipid mediators. Targeting lipid signaling might offer new therapeutical targets for improving the clinical outcome in cardiovascular disease patients. We considered lysophosphatidylinositol (LPI) and its receptor G protein-coupled receptor (GPR)55 as a potential modulator of atherosclerosis. Its role in regulating atherosclerosis and B cell function is unknown.
We hypothesize that GPR55 signaling affects atherosclerosis by regulating B cell function.
Atherosclerotic plaques were compared between apolipoprotein-E-deficient (ApoE−/−) and ApoE−/−Gpr55−/− mice after 4 to 16 weeks Western Diet (WD; 0.15% cholesterol; n=12–15 per group). To test the role of B cell GPR55, we generated mixed chimeras by irradiating low density lipoprotein receptor deficient (Ldlr−/−) mice and reconstituting with a mixture of μMT and wildtype or μMT and Gpr55−/− bone marrow cells. Circulating B cells were sorted and bulk RNA sequencing analysis was performed. We performed atheroma plaque characterization, qPCR and ELISA of tissue lysates and measure plasma immunoglobulins. Circulating and tissue leukocyte counts were determined.
We confirmed Gpr55 expression on circulating B cells, which was higher compared to T and myeloid cells. ApoE−/−Gpr55−/− mice had significantly larger plaques after 4 & 16 weeks WD compared to ApoE−/−, with increased body weight & cholesterol levels. In addition, global Gpr55 deficiency resulted in enhanced aortic pro-inflammatory cytokine mRNA expression, a massively upregulated IgG levels and increased counts of splenic germinal center and plasma cells. ApoE−/−Gpr55−/− B-cell RNA-seq analysis showed 460 differential expressed genes compared to ApoE−/−. The main pathways affected were calcium ion transport, immunoglobulin production, T & B cell activation, and cellular response to stress. B cell specific Gpr55 deficiency blunted the metabolic effects but still translated in larger atherosclerotic plaques and elevated plasma IgG levels.
Both global and B cell-restricted Gpr55 deficiency promotes atherosclerosis and is associated with a pro-inflammatory phenotype.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): The authors received funds from the Deutsche Forschungsgemeinschaft (STE1053/6-1, STE1053/8-1 to S.S. and SFB1123 to S.S., C.W. and L.M.), the German Ministry of Research and Education (DZHK FKZ 81Z0600205 to S.S.) and the LMU Medical Faculty FöFoLe program (1061 to R.G.P.). I.H. is supported by the DFG (HI1573/2 and CRC1425 #422681845).