bluuebunny/crossref_metadata_2025_split huggingface dataset:
DNMT3A CHIP-driver mutations ignite the bone marrow
Authors: Dederichs T S , Ehlert C , Becker H , Wolf D , Czerny M , Jaiswal S , Westermann D , Hilgendorf I 
Published: October 2024

Abstract

Background
Clonal hematopoiesis of indeterminate potential (CHIP) is an acquired, genetic cardiovascular (CV) risk factor affecting about 10% of the general population at the age of 70 years. Most CHIP-driver mutations affect the epigenetic regulator gene DNMT3A. Experimental studies suggest a causal link between CHIP and inflammation-driven atherosclerosis, but the underlying mechanisms of how few mutant clones provoke adverse clinical outcomes in patients remain obscure.

Methods
Single monocytes and bone marrow cells from DNMT3A-mutation carriers undergoing coronary angiography and open-heart surgery underwent RNA and genomic DNA sequencing in parallel on a single cell level. This approach enabled us to compare mutant and non-mutant cells directly within carriers, and to non-carrier cells. Atherosclerotic chimeric mice reconstituted with a mixture of Dnmt3a-deficient (CD45.2) and -competent (CD45.1) bone marrow served as an experimental model to compare mutant to non-mutant macrophages in atherosclerotic lesions.

Results
Surprisingly, RNA/gDNA parallel sequencing revealed that gene expression profiles of mutant monocytes resembled those of non-mutant monocytes in carriers of DNMT3A-CHIP mutations (n=4). Collectively, however, monocytes of DNMT3A mutation carriers were more inflammatory than those of matched non-carrier patients. Mutant hematopoietic stem and progenitor cells, analyzed by RNA/gDNA parallel sequencing in two carriers, expressed more activation and inflammatory markers compared to non-mutant cells within the same bone marrow, and intraindividual differences declined as cells differentiated into monocytes. Macrophages were isolated from atherosclerotic aortas of mixed bone marrow chimeric mice based on CD45.1 (non-mutant) or CD45.2 (mutant or non-mutant controls) expression and subjected to single cell RNA sequencing. Inflammation markers were higher expressed in non-mutant macrophages within carrier mice than in non-mutant macrophages of non-carrier chimeras, and showed relatively small differences compared to Dnmt3a-deficient macrophages.

Conclusions
Our experimental findings in humans and mice support a pathomechanistic model in which few DNMT3A-CHIP mutation-carrying hematopoietic stem cells stimulate non-mutant cells within the bone marrow, thereby augmenting inflammation downstream in all circulating monocytes and their progeny in atherosclerotic lesions.
The orchestration of coding and non-coding genes at the CDH13 locus in coronary artery disease
Authors: Chen Z , Li S , Li L , Diagel A , Moggio A , Li Z , Song X , Sager H , Boon R , Maegdefessel L , Kessler T , Von Scheidt M , Bjorkegren J L M , Maliheh N J , Schunkert H 
Published: October 2024

Abstract

Background
The CDH13 locus was associated with coronary artery disease (CAD) by genome-wide association studies (GWAS). However, the causal gene(s) and underlying disease mechanisms at this locus remain undetermined.

Purpose
We intended to pinpoint and functionally validate the causal coding and non-coding genes at the CDH13 locus and delineate the underlying mechanisms for CAD.

Methods and results
We conducted the transcriptome-wide association analysis (TWAS) using nine types of CAD-relevant tissues from Stockholm-Tartu Atherosclerosis Reverse Network Engineering Task (STARNET) and Genotyped Tissue Expression (GTEx) projects and identified CDH13 (T-cadherin) and four long non-coding RNA (lncRNA) genes as candidate causal genes at The CDH13 locus. One of the antisense(AS) lncRNAs, CDH13-AS2, was predicted to bind on the 3’ untranslated region (UTR) of CDH13 using LncRRIsearch and RNAup servers. The binding interaction was validated by dCas13-medicated RNA immunoprecipitation. Weighted correlation network analysis (WGCNA) using transcriptomic data from CAD-relevant tissues of ~600 human subjects suggested a positive expression correlation between CDH13 and CDH13-AS2 in artery tissues. CRISPR/Cas9-based knockout (KO) of CDH13 or CDH13-AS2 in human umbilical vein endothelial cells (HUVECs) indicated synergistically atherogenic phenotypes, including reduced proliferation and migration, and increased apoptosis and monocyte adhesion. The data matched with the downregulation of CDH13 in artery samples of atherosclerosis patients. The cellular phenotypes were also in line with our in vivo data of Cdh13 and Apoe double-KO (Cdh13-/-/Apoe-/-) mice, which showed increased aortic lesion areas when fed on a high-fat diet compared to Apoe-/- mice. Furthermore, prediction using TargetScan, miRWalk, and scanMiRApp databases identified the potential binding of several EC-expression microRNAs (mir) on 3’UTR of CDH13. Several disease-associated variants were mapped to the predicted binding sites of a few microRNAs, including mir-let7, mir-30, and mir-125. The dual luciferase-based RNA interference (RNAi) assay confirmed the binding of mir-let7 on 3’UTR of CDH13. Transcriptional activation of CDH13-AS2 by the enzymatically inactivated Cas9 (dCas9) system diminished the binding of mir-let7 and increased CDH13 expression.

Conclusions
At the CDH13 locus, CDH13 and CDH13-AS2 are two causal genes, whose downregulation likely contributes to CAD. CDH13-AS2 bound and stabilized CDH13 partially by preventing mir-let7 mediated degradation of CDH13 mRNA, suggesting therapeutic potentials.
Comparing cerebral microembolisation patterns between high power short duration and pulsed field ablation techniques using artificial intelligence based robotic transcranial doppler monitoring
Authors: Meszaros H , Vamosi P , Komlosi F , Toth P , Szegedi N , Sallo Z , Perge P , Geller L , Simon Z S , Nardai S , Nagy K V 
Published: October 2024

Abstract
Thromboembolism is a rare (&lt;1%) complication of atrial fibrillation (AF) ablation , but the incidence of silent ischaemic lesions (SIL) is 5-15% (1). Artificial intelligence (AI)-based robotic transcranial Doppler (r-TCD) enables real-time cerebral embolisation detection.
We aimed to investigate the cerebral microembolization signal (MES) load with r-TCD of patients undergoing PF ablation, comparing the number of detected MES-s between 90 W radiofrequency (RFA) "very high power short duration (vHPSD) ablation" and "pulsed field ablation (PFA)" techniques.
Our interim analysis included r-TCD registratums of 26 patients. 16 interventions were performed with VHPSD, 10 with PFA, during which we recorded the flow curve of the arteria cerebri media (ACM) bilaterally with r-TCD and analyzed the MES-s quantitatively and qualitatively with AI-based embolisation detection software. Postoperative cranial MRI exams were performed. Continuous variables were compared between the groups using a two-sample t-test, and categorical variables were compared using a Fisher exact test. The MES load by the different pulmonary veins was compared using Wilcoxon test and Bonferroni correction. A p-value below 0,05 was considered significant.
The two groups were comparable regarding baseline parameters (mean age: 63 vs.58 p=0,253; EF:59% vs.62 .2% p=0,33). Ablations with vHPSD were significantly longer compared to PFA (88,7 min vs. 54,7 min p&lt;0,001), left atrial time was significantly shorter with PFA (52,1 min vs. 29 min p&lt;0 001), but the fluoroscopic time was significantly longer (5,4 min vs. 9,3 min p=0,009) during ablation with PFA. Neither solid emboli, nor ischaemic lesion or neurological deficit was detected. During the ablations, we saw an average of 246 gas MES-s with PFA and 138 with RFA (p=0,216). During PFA we analyzed the MES load per pulmonary vein: by the left superior vein we detected a median of 56,5 (IQR: 21,5-116,5), by the left inferior there were a median of 23 MES-s (IQR: 4,5 115,5), by the right inferior a median of 32 (IQR: 12-84,5), and by the right superior a median of 22,5(IQR: 8,5-70) respectively, p=0,03. While comparing each veins to each other, by the right vs. left superior vein, the p value resulted 0,01.
On an initial sample,we detected known differences in the procedural parameters of the two interventions. There tended to be more MES-s during PFA, but this difference was not significant. Regarding the MES load by the ablation of different pulmonary veins, we found a significant overall difference between the MES load by the four veins, especially between the left and right superior vein. A larger number of interventions and r-TCD registratums is needed to determine the cerebral safety of PFA compared to RFA.
Blockade of foam cell formation by an AP-1 inhibitor attenuates atherosclerosis in diabetes
Authors: Khan A W , Aziz M , Sourris K , Jandeleit-Dahm K A M 
Published: October 2024

Abstract

Background
Atherosclerosis is a major contributor to cardiovascular disease (CVD) related premature deaths in people with diabetes. Residual increased risk of CVD remains even after treatment of standard risk factors including hyperglycaemia. Therefore, there is an unmet clinical need of novel therapies that can directly target the intrinsic pathobiology of atherosclerotic plaque development. Macrophage associated pro-atherosclerotic processes including impaired cholesterol efflux resulting in foam cell formation are important therapeutic targets in atherosclerosis. Expression of genes linked to impaired cholesterol efflux can be regulated by multiple factors including transcription factors (TFs) such as activator protein 1 (AP-1) complex. Although, previous studies have implicated the role of foam cells in atherosclerosis, the role of AP-1 TF complex in foam cell formation in diabetes induced atherosclerosis is not known.

Methods
Diabetes was induced with streptozotocin (STZ) in atheroprone Apoe-/- mice and 10 weeks later aortae were subjected to single cell RNA sequencing (scRNA-seq) analysis using 10X Genomics platform. In vitro model mimicking diabetes-induced foam cell formation was established in human monocytes THP-1 derived macrophages using high glucose (25mM) and ox-LDL ± a small molecular inhibitor of AP-1, T-5224. AP-1 activity, gene expression and foam cell formation were assessed with activity assay, RT PCR and Oil Red O staining respectively in this setting. Complementary in vitro AP-1 knockdown experiments using shRNA were also performed. Furthermore, preclinical intervention studies were also performed where Apoe-/- mice made diabetic with STZ were treated 5-weeks post diabetes with T-5224 (30 mg/kg body weight daily for 5 weeks).

Results
scRNA-seq analysis identified a well-defined cluster of foam cells within macrophages with a diabetes specific proatherogenic transcriptomic profile. Differential gene expression analysis showed increased expression of AP-1 members cFos, Atf3, Atf4 and Junb exclusively in foam cell cluster in diabetes (cut off = FDR&lt;0.05, Log2FC -2/2). Comparison with the ENCODE dataset for AP-1 target genes using Harmonizome version 3 showed that indeed multiple differentially expressed genes in diabetes were targets of AP-1. AP-1 members were also upregulated in the THP-1-based in vitro model of high glucose induced foam cell formation. THP-1 derived foam cells showed increased AP-1 activity and lipid uptake. Interestingly, AP-1 inhibition by T-5224 attenuated foam cell formation. AP-1 knockdown experiments validated experimental findings of AP-1 inhibition studies in THP-1 derived macrophages. Importantly, T-5224 treatment blocked foam cell formation resulting in reduced atherosclerosis in diabetic Apoe-/- mice in our intervention study design.

Conclusion
This study identified the AP-1 inhibition with T-5224 as a potential treatment for the diabetes induced foam cell formation and associated atherosclerosis.
Diesel exhaust particles activate macrophages and IRF5 expression in atherosclerosis
Authors: Seneviratne A , Park I , Dib L , Mudway I , Udalova I , Miller M , Monaco C 
Published: October 2024

Abstract

Introduction
Over 20% of global deaths related to cardiovascular disease have been attributed in part to air pollution, particularly fine particulate matter (PM2.5) (1). In highly trafficked urban areas, smaller ultrafine diesel exhaust particles (DEP) represent a significant proportion of PM2.5. DEP is associated with the progression of atherosclerosis and increased plaque susceptibility to rupture (2). We have previously shown that interferon regulatory factor 5 (IRF5) has a key role in necrotic core formation, a distinguishing feature of vulnerable atherosclerotic plaques, by rendering macrophages defective at efferocytosis (3). Pronounced elevation of IRF5 is also seen in symptomatic carotid plaques, particularly in the rupture-prone shoulder regions (4).

Purpose
We examined the effect of DEP on macrophage phenotype and IRF5 expression in atherosclerosis.

Methods
Bone marrow-derived macrophages (BMDMs) from C57BL/6 mice, differentiated with GM-CSF (to mimic inflammatory macrophages) or M-CSF (mimicking homeostatic resident macrophages) were incubated with PBS or DEP (10 µg/mL) in vitro for 18 hours. Expression of inflammatory cytokines (IL-6, IL-12α), macrophage polarisation markers CD11c (itgax, inflammatory) and CD206 (mrc1, resident anti-inflammatory) and IRF5 was quantified by real-time PCR. In addition, high fat-fed ApoE-/- mice were exposed to saline or DEP (35 µL at 1 mg/mL) twice weekly for 5 weeks by pulmonary administration. The pan-macrophage marker CD64, CD206 and IRF5 were quantified in paraffin-embedded sections of brachiocephalic artery lesions by immunohistochemistry.

Results
In M-CSF-differentiated murine macrophages incubated with DEP, significant fold increases in gene expression (relative to PBS control) were observed: IRF5 (1.4-fold), IL-6 (31.5-fold) and IL-12α (5.8-fold) while CD206 (0.8-fold) decreased (Figure 1a, n = 5, p&lt;0.05, two-tailed paired t-tests). CD11c expression was unchanged. No statistically significant differences were detected in GM-CSF-differentiated macrophages. In the brachiocephalic arteries of DEP-exposed ApoE-/- mice we observed increases in CD64+ macrophages (28.2±7.4 vs 15.5±5.6) and IRF5 (16.6±3.6 vs 10.1±3.4) expression, while CD206 expression decreased (4.1±0.9 vs 11.0±1.2) in comparison to saline-exposed mice (Figure 1b mean±SD, n = 5, p&lt;0.05, two-tailed unpaired t-tests, data expressed as % of intima area).

Conclusion
DEP promotes CD64+ macrophages and IRF5 expression in murine atherosclerosis, while inflammation and IRF5 is induced in vitro, but only in M-CSF-differentiated murine BMDMs. This suggests that DEP has no impact on inflammatory macrophages but could switch homeostatic resident macrophages to a more inflammatory phenotype via the activation of IRF5. Therefore, IRF5 is a promising candidate for further investigation into the initial macrophage response to inhaled particles that leads to the exacerbation of vascular disease.
Digoxin reduces senescence and restores dysfunctional endothelial-subcutaneous adipocyte cross-talk and dysregulated glucose homeostasis in patients with type 2 diabetes mellitus
Authors: Brown O I , Bridge K I , Straw S , Luk C , Makava N , Tayebjee M , Saunderson C E D , Bruns A F , Drozd M , Gierula J , Wheatcroft S B , Witte K K , Cubbon R M , Kearney M T 
Published: October 2024

Abstract

Background
Normal subcutaneous adipose tissue (SAT) function is dependent on SAT microvascular expansion. Heart failure with reduced ejection fraction (HF) and type 2 diabetes mellitus (DM) have a synergistic deleterious relationship and, of relevance to this, metabolic dysregulation may drive HF progression in DM. Chronic hyperinsulinemia is associated with adipocyte senescence, however any contributing role of SAT microvascular endothelial cell (SATMVEC) senescence on adverse microvascular expansion in type 2 diabetes mellitus (DM) remains unknown. Digoxin has been shown to have senolytic activity in murine cancer models. However, the potential therapeutic role of digoxin on SAT metabolic function is unexplored.

Purpose
We establish the presence of SATMVEC senescence and its effects on adipocyte function in people with HF and DM (HFDM). Moreover, we establish a therapeutic role for digoxin in targeting SATMVEC senescence, modulating SATMVEC-adipocyte crosstalk and restoring metabolic homeostasis.

Methods
We took pectoral SAT biopsies from 59 patients undergoing pacemaker implantation of whom 25 (42%) had HF and DM (HFDM). SATMVEC were isolated from SAT and were treated with 1ng/mL digoxin vs control for 24 hours before characterisation (cell function, expression of senescence-associated ß-galactosidase (ß-gal) and senescence associated secretory phenotype, SASP). Crosstalk between SATMVEC and subcutaneous white adipocytes were examined using co-culture techniques (Fig 1A). Paired digoxin vs control treated SATMVEC were seeded co-cultured with adipocytes for 24 hours, before adipocyte health was quantified, through 2-NBD-glucose uptake and expression of key adipogenic genes and proteins.

Results
Compared to HF alone, isolated SATMVEC from patients with HFDM had a senescent phenotype with increased SASP and ß-gal expression (p&lt;0.05), reduced metabolic activity, ATP production, proliferative capacity, and impaired angiogenesis (p&lt;0.05). In co-culture, SATMVEC from patients with HFDM had deleterious effects on adipocyte function: increasing expression of IL-6 (Fig 1C) and reducing 2-NBD-glucose uptake (p&lt;0.05). Treatment of SATMVEC with digoxin reduced the SATMVEC senescent phenotype (Fig 1B) and increased 2-NBD-glucose adipocyte uptake (Figure 1D) (p&lt;0.001).

Conclusions
SATMVEC from patients with HFDM have a senescent phenotype, and when in co-culture induce a proinflammatory adipocyte phenotype, resistant to glucose uptake. Digoxin reduces SATMVEC senescence, restores healthy adipocyte glucose homeostasis, and thus has the potential to repair dysfunctional SAT in patients with HFDM.
Deletion of the Uox gene generates a mouse model of hyperuricemia with cardiovascular complications
Authors: Shali S , Zeng L , Gao Y B , Zhou P , Dai Y X 
Published: October 2024

Abstract
Background/introductions: Hyperuricemia is associated with hypertension, coronary atherosclerosis and heart failure. However, the causal relationship between uric acid (UA) and cardiovascular disease remains controversial. There has been lack of mouse model for spontaneous hyperuricemia with cardiovascular complications.

Purpose
We aimed to develop urate oxidase (Uox) deficient mice with hyperuricemia that can survive to adulthood.

Methods
Uox-knockout mouse model was generated on C57BL/6J background by deleting exon 2-4 of Uox using the CRISPR/Cas9 system. we evaluated the hyperuricemic phenotypes regarding magnitude of serum UA elevation, cardiometabolic abnormalities, and cardiovascular complications.

Results
The birth rate of homozygous Uox-knockout (Uox -/-) mice was 21.4%. The prototypic Uox -/-mice had 5.5-fold increased serum uric acid (1351.04±276.58 μmol/L) as compared to the wild type mice (P&lt;0.0001)), but mostly died by four weeks. After allopurinol (3ug/g) intervention, they all survived more than eight weeks (median: 12.6 weeks, range:9.6 to 16.0 weeks). The serum UA was 612.55±146.98μmol/L (2.5 times higher than that in WT mice, P&lt;0.0001) in the eight-week-old allopurinol-rescued Uox -/-mice, which also manifested multiple complications including hypertension (systolic blood pressure:155.25±9.48mmHg vs. 109.53±11.02mmHg, P&lt;0.0001), left ventricular remodeling and systolic dysfunction (all echocardiographic functional parameters, P&lt;0.001), aortic endothelial cell impairment (P&lt;0.001 for proliferating cell nuclear antigen and ZO-1, P=0.027 for VE-cadherin), hepatic steatosis and elevated liver enzymes (all enzymes P&lt;0.0001), as well as urate nephropathy, hyperglycemia (P=0.0146 for fasting glucose) and hypercholesteremia (P=0.0045 for total cholesterol).

Conclusion
The present Uox-/- mice developed spontaneous hyperuricemia complicated with cardiovascular disease and cardiometabolic disorders. This allopurinol-rescued Uox-/- mouse model could survive to adulthood, and may provide a novel tool to study urate biology and hyperuricemia associated early-onset disorders in human.
Figure 1 Figure 2
Semaglutide modulates pro-inflammatory neutrophil phenotype induced by supraphysiological levels of the adipokine FABP4 in patients with cardiovascular disease
Authors: Sanchez-Lopez D , Garcia-Vega D , Basdas R , Fu X , Nunez-Caamano J R , Al-Rubaye S , Vinuela-Roldan J E , Reija-Lopez L , Martinez-Cereijo J M , Fernandez-Gonzalez A L , Gonzalez-Juanatey J R , Eiras-Penas S 
Published: October 2024

Abstract

Background
Increased adiposity is a risk factor for cardiovascular disease (CVD). Adipose tissue is an endocrine organ that releases proteins with pro-inflammatory effects. One of them is FABP4, which levels are increased after adipogenesis. Insulin resistance is linked to supraphysiological levels of this biomarker. Therapies to reduce body weight and improve insulin response, such as the GLP-1 receptor agonists, like semaglutide, have been shown to benefit patients with CVD and obesity, who have a high pro-inflammatory neutrophil profile.

Purpose
To study the FABP4 effects on neutrophils from patients with cardiovascular disease and cellular models and its possible modulation by semaglutide.

Methods
We included 11 patients undergoing open heart surgery. Neutrophils from blood were isolated by single-step centrifugation, counted, and same number were treated with FABP4 (100 ng/mL), semaglutide (1nM) or both. Finally, their phenotype was assessed using flow cytometry (FACS). The same treatment conditions were used on differentiated HL-60 cells (dHL-60) to test their phenotype and functionality based on a) oxidative burst capacity using dihydrorhodamine 123 and b) adhesion to coronary artery endothelial cells. At the end, neutrophils phenotype was tested on patients with obesity and diabetes under semaglutide treatment for 6 months, according to clinical practice. Neutrophils phenotype was determined by FACS and FABP4/C5a plasma levels by immunofluorescence assay.

Results
FABP4 reduced neutrophils’ CD88 (p=0.006) and CXCR2 (p=0.02) levels. This was also verified in dHL60 cells (p=0.01). In these cells, we were also able to observe a CD11b increment after FABP4 treatment (p=0.04), modulated by semaglutide (p=0.02). This phenotype increased the neutrophil-coronary adhesion (p=0.03). Their oxidative burst capacity was attenuated after semaglutide treatment. In addition, 32% of patients underwent semaglutide treatment reduced plasmatic FABP4 (p=0.03) levels and increased CD88 (p=0.01). We confirmed that CD88 modulation does not lead to a change in plasma C5a.

Conclusions
High FABP4 levels reduce CD88 of neutrophils from patients with cardiovascular disease and enhance their adhesion to coronary endothelial cells. Modulation of the FABP4 effects by semaglutide treatment might suggest a protective mechanism against adiposity/coronary endothelial inflammation.