bluuebunny/crossref_metadata_2025 huggingface dataset:
Single-nucleus RNA sequencing of TMEM43 induced arrhythmogenic cardiomyopathy reveals altered cell compositions and transcriptional states in mice
Authors: Lalaguna L , Lopez-Olaneta M , Villalba-Orero M , Garcia-Pavia P , Lara-Pezzi E
Published: November 2023

Introduction and purpose

TMEM43 induced Arrhythmogenic cardiomyopathy, also referred to as arrhythmogenic cardiomyopathy type 5 or ACM5, is caused by a point mutation in TMEM43 (p.S358L). The disease is characterised by chamber dilation and a progressive dystrophy of the ventricular myocardium causing life-threatening ventricular arrhythmias and sudden cardiac death. Loss of cardiomyocytes and fibrofatty replacement of the myocardium is clearly observed in our transgenic mouse model of ACM5 overexpressing TMEM43-S358L in postnatal cardiomyocytes. However, the mechanisms underlying cardiomyocyte death and fibroblast activation remain elusive, precluding the development of effective therapeutic options. Here we applied the state-of-the-art analysis on single-nucleus RNA sequencing (snRNA-seq) to characterise the cellular compositions and molecular states in ACM5 mouse hearts.

Methods

Left ventricular heart tissues of severely affected ACM5 mice and their corresponding transgenic controls overexpressing wild-type TMEM43 were used for snRNA-seq analysis. We identified 10 major cell types, which aggregated into 23 distinct clusters on the basis of transcriptional similarity.

Results

ACM5 nuclei showed significant depletion of cardiomyocytes and expanded populations of fibroblasts and myeloid cells. Fibroblasts from mutant mice acquired an activated phenotype related to remodelling of the extracellular matrix with a large number of genes like LOX, POSTN, NOX4 or COL1A1 significantly upregulated. In addition, the mutant cardiomyocytes showed altered transcriptional states characterized by cardiac stress and heart failure markers such as NPPB or MYH7. Interestingly, we were able to identify a cluster of mutant cardiomyocytes with a highly apoptotic profile. This cluster is also enriched in genes related to the unfolding protein response (UPR) stress sensors and the DNA damage response (DDR), elucidating a new potential mechanism of myocyte death in ACM5. Furthermore, heterogeneity of the myeloid populations is also observed. The number of tissue resident macrophages is increased in mutant samples and myeloid cells show additional disease-associated transcriptional states that are almost entirely absent in control samples.

Conclusion

Together these data illuminated shared and distinct cell and molecular architectures of ACM5 in mouse hearts. Further confirmation on diverse gene expression profiles like the UPR and DDR pathways related to cardiomyocyte death will establish a valuable resource to explore novel therapeutic targets and personalized therapies for ACM5.
Unraveling the pathogenesis of novel ANKRD1 missense mutation causing familial dilated cardiomyopathy using human-induced pluripotent stem cells derived cardiomyocytes
Authors: Ballan N , Shaheen N , Ghiringhelli M , Carmon T , Shiti A , Arabel G , Gepstein A , Huber I , Gepstein L
Published: November 2023

Introduction

Familial dilated cardiomyopathy(f-DCM) can be caused by mutations in 30 different genes encoding sarcomeric, cytoskeletal, mechanotransducers and transcription factors. One of the genes ANKRD1 encode Cardiac ankyrin repeat protein (CARP) which is involved in the mechanosensory unit located on the I-band of the sarcomere. Here we demonstrate a novel mutation in ANKRD1 gene causing f-DCM, and the usage of human-induced pluripotent(hiPSC) technology unraveling the pathogenesis of the disease.

Purpose

of the study:

1) Establishing patient specific hiPSC-derived cardiomyocytes from a family harboring a mutation in ANKRD1 gene causing f-DCM; 2) Phenotypical assessment of the morphological and ultrastructural properties of the diseased cells upon CARP activation; 3) Functional assessment of the 3 component of excitation-contraction coupling at the single-cell and tissue level.

Methods and Results

Patient-specific hiPSC were generated from a family harboring missense mutation in the ANKRD1 gene, with a substitution of the amino acid arginine to cysteine at position 182 of the Cardiac ankyrin repeat protein (CARP). ANKRD1-mut hiPSC were differentiated into cardiomyocytes and later matured using hormonal treatment. Morphological assessment using immunofluorescence(IF) techniques revealed that ANKRD1-mut hiPSC-CMs demonstrate an attenuated response to hypertrophy (upon Angiotensin-2 and Phenylephrine treatment). In addition ANKRD1-mut hiPSC-CMs displayed a higher cellular death rate upon increasing substrate stiffness and starvation. Electrophysiological assessment of the ANKRD1-mut cardiac tissues exhibit no alternations in action potential durations and conduction velocities while assessment of calcium transients demonstrated slower kinetics. In addition forces were quantified at the single-cell and tissue level , and demonstrated reduced active tension values and abnormal post-extrasystolic potentiation, while only single-cell experiments have demonstrated decreased passive tensions. These phenotypical alternations might be attributed to decreased nuclear translocation of CARP upon phenylephrine treatment as detected in IF experiments.

Conclusion

Novel mutation in ANKRD1 were found and validated to be causing f-DCM, by revealing phenotypical alternation using hiPSC-CM technology. Increased cellular death rate and impaired nuclear translocation of CARP might be the underlying causes of the morphological and contractile abnormalities detected in-vitro, leading to systolic dysfunction in the affected family.
N-terminal pro-B-type natriuretic peptide for prediction of ventricular arrhythmias: data from the SMASH Study
Authors: Sourour N , Riveland D R , Nasgaard D R , Kjekshus P H D , Larsen P H D , Omland P H D , Rosjo P H D , Myhre P H D
Published: November 2023

Background

Elevated N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentrations predict heart failure (HF) and mortality, but whether NT-proBNP predicts ventricular arrhythmias (VA) is not clear.

Purpose

We hypothesize that high NT-proBNP concentrations associate with the risk of incident VA, defined as adjudicated ventricular fibrillation or sustained ventricular tachycardia.

Methods

In a prospective, observational study of patients treated with implantable cardioverter defibrillator (ICD), we analyzed NT-proBNP concentrations at baseline and after mean 1.4 years in association to incident VA.

Results

We included 491 patients (age 66±12 years, 83% men) out of whom 51% had a primary prevention ICD indication. The median NT-proBNP concentration was 567 (25-75 percentile 203-1480) ng/L, and patients with higher concentrations were older with more HF and ICD for primary prevention. During mean 3.1±0.7 years, 136 patients (28%) had ≥1 VA. Baseline NT-proBNP concentrations were associated with risk of incident VA (HR 1.39 [95%CI 1.22-1.58], p<0.001) (Figure1), HF hospitalizations (HR 3.38 [95%CI 2.69-4.25], p<0.001) and all-cause mortality (HR 2.49 [2.05-3.04], p<0.001) (Figure2), which persisted after adjusting for age, sex, BMI, CAD, HF, renal function, and LVEF. The association with VA was stronger in secondary vs primary prevention ICD indication: HR 1.59 (95% CI 1.34-1.88 C-statistics 0.71) vs HR 1.25 (1.02-1.52, C-statistics 0.55), p-for-interaction=0.06. Changes in NT-proBNP during the first 1.4 years did not associate with subsequent VA.

Conclusions

NT-proBNP concentrations are associated with the risk of incident VA after adjustment for established risk factors, with the strongest association in patients with a secondary prevention ICD indication.
RNA surveillance by ADAR1 is required for vascular endothelial resilience
Authors: Tual-Chalot S , Ciliberti G , Polycarpou-Schwarz M , Amponsah-Offeh M , Turchinovich A , Dimmeler S , Gatsiou A , Stellos K
Published: November 2023

Introduction

Long dsRNAs are recognised as danger-associated molecular patterns by the cytosolic innate immune sensors inducing thus innate immune responses. RNA editing is a post-transcriptional modification induced by the binding of Adenosine Deaminases Acting on RNA (ADARs) to double-stranded RNA structures (dsRNAs). We have recently reported that ADAR1 is the main RNA editor in endothelial cells (ECs) but the role of ADAR1 in vascular homeostatic processes remains yet elusive.

Methods

Genetic deletion strategies were employed by crossing mice carrying a conditional (floxed) Adar1 , Ifh1 or Tlr3 allele with either a Tie2-Cre or with a tamoxifen-inducible VE-Cadherin-CreERT2 mouse line. Primary human and murine vascular EC culture assays, dsRNA metabolism studies, gene-silencing and expression techniques, immunohistochemistry and confocal microscopy were used to assess the EC-specific ADAR1 effects.

Results

EC-specific ADAR1 deletion resulted in prenatal embryonic lethality evidencing an absolute requirement of EC-ADAR1 in life. Inducible EC-ADAR1 ablation in adult mice elicited premature sudden death due to a widespread lung vascular leakage and pleural effusion, reflecting a compromised EC barrier function. Mechanistically, silencing of endothelial ADAR1 disrupted long-to-short dsRNA metabolism witnessed by the striking accumulation of cytoplasmic long dsRNAs. Subsequently, the cytoplasmic accumulation of endogenous long dsRNAs induced aberrant levels of interferon-β and activation of innate immune system. Finally, activation of the cytosolic dsRNA sensors inflicted the dissociation of β-catenin from VE-cadherin in EC junctions, while, co-silencing of the dsRNA sensor, MDA5, was sufficient to restore the integrity of the ADAR1-deficient endothelium. While double EC-specific ADAR1 and TLR3 did not fully rescue the lethality of the EC-ADAR1 animals, in vivo experiments using double EC-specific ADAR1 and MDA5 KO mice fully rescued the phenotype. Validation experiments using murine vascular ECs isolated from the mouse strains of our studies confirmed our mechanistic and phenotypic findings.

Conclusion

Endogenous suppression of double-stranded RNA-induced MDA5 autoinflammatory signaling by the RNA editor ADAR1 is essential for the maintenance of vascular integrity in adult mice.
Bradycardic effect of endothelial-specific bradykinin type-II receptors overexpression in mice: impact on blood pressure
Authors: Metry S , Suvorava T , Kojda G
Published: November 2023

Background/Introduction

Polymorphisms of the gene encoding for the bradykinin type-II receptors (B2R) has been shown to decrease the expression of B2R and increased vascular resistance and blood pressure in humans.

Purpose

We aimed to study the role of endothelial B2R on systolic blood pressure (SBP) and heart rate (HR) using a novel mice model with endothelial-specific B2R overexpression (B2tg) as compared with transgene negative littermates (B2n) and on differential expression of proteins potentially relevant to vascular tension.

Methods

Human and murine B2R expression levels were quantified by rtPCR. Thoracic aortic tissues were used for vascular reactivity studies and proteomic analysis. SBP and HR were measured by the tail-cuff method at baseline, after administration of the COX inhibitor diclofenac (10 mg/kg/day), B2R antagonist icatibant (1 mg/kg) or NOS inhibitor L-NAME (100 mg/kg/day).

Results

In aortic tissues of B2tg human B2R were exclusively expressed (n=6, P=0.0005), while murine B2R expression levels remained unchanged. Similar results were found in skeletal muscle tissues used as an estimate for resistance vessels. Aortic ring studies revealed a concentration-dependent constriction to bradykinin in wild-type mice (maximal vasoconstriction 9.6±3.0%, n=5) whereas aortic rings of B2tg showed concentration-dependent relaxation (-27.4±4.1%, n=12). SBP was significantly lower in B2tg (125.0 ±1.6 mmHg) as compared to B2n (129.9±1.1 mmHg, n=9, P=0.03). Diclofenac treatment slightly but not significantly reduced SBP in B2n (to 125.5±2.4 mmHg, P=0.09) and in B2tg (to 119.8±2.8 mmHg, n=9, P=0.12), however the difference in SBP remained (P=0.03). In a pilot study, L-NAME administration did also not change the SBP difference between B2n and B2tg (9.9±10.3 mmHg, n=3-4). In contrast, icatibant administration abolished the difference between the groups (B2n 128.6±4.6 vs B2tg 128.3±2.0 mmHg, n=8, P=ns) suggesting the involvement of endothelial B2R in SBP reduction. Interestingly, in the same mice, HR was significantly lower in B2tg (571.4±20.8 bpm) as compared to B2n (626.7±13.7 bpm, P=0.02). While diclofenac didn’t affect HR, icatibant significantly increased HR to (630.8±21.8 bpm in B2tg and 685.5±16.4 bpm in B2n, n=8, P=0.01). Only in B2tg, there was a significant strong correlation between SBP and HR (r=0.76, P=0.016). Analysis of proteomic data revealed differential upregulation of proteins related to cell adhesion (CD44), extracellular matrix (hyaluronidase), and differential downregulation of proteins related to oxidative stress (superoxide dismutase) in B2tg.

Conclusion

Endothelial B2R overexpression reduces SBP and HR. SBP reduction is not dependent on two major vascular pathways stimulated by products of NOS and COX but appears to be related to endothelial B2R-dependent HR reduction. Hence, agonism at B2R could be a valuable target for development of new cardiovascular drugs.
B cells and immunoglobulins promotes aortic dissection in mice
Authors: Ohno S , Aoki H , Furusho A , Hashimoto Y , Nakao E , Shibata R , Nishida N , Ito S , Majima R , Fukumoto Y
Published: November 2023

Background

Aortic dissection (AD) is a devastating aortic disease with high mortality, for which molecular pathogenesis is largely unknown. Recent studies have highlighted the importance of inflammatory response in AD. We have reported that B cells and immunoglobulins promote inflammatory response in the aorta and are involved in the pathogenesis of abdominal aortic aneurysm, another form of aortic disease. However, it is not known whether and how B cells participate in AD development or progression.

Methods and results

Immunohistochemical staining of human AD tissue revealed that B cells were clustered together with T cells, macrophages, and neutrophils at the entry site of AD with medial disruption. B cell cluster was also observed at the site of medial disruption in mouse model of AD that was induced by continuous infusion of beta-aminopropionitrile and angiotensin II (BAPN+AngII). In muMT mouse, which is genetically deficient for B cells and immunoglobulins, BAPN+AngII induced significantly less severe AD compared to that in wild type. The rate of aortic rupture and sudden death was approximately 42% in wild type mice, while that in muMT mouse was 12% (P<0.05). Administration of mouse normal polyclonal IgG to muMT mice resulted in dramatic increase in aortic rupture and sudden death, starting at day 7 of BAPN+AngII infusion, and reaching 69% of rupture rate, indicating the critical role of IgG in AD. Transcriptome analysis revealed that IgG administration enhanced the expression of inflammation-related genes in muMT mice before BAPN+AngII infusion, indicating that IgG increased susceptibility to AD. Immunohistochemistry of aorta confirmed exogenous IgG deposition, colocalized with C3, which is known to contribute to innate immunity.

Conclusion

These findings demonstrated B cells and IgG are critically involved in the destructive inflammation of AD pathogenesis. Further, the aortic deposition of IgG and C3, a part of the components of innate immunity, precedes the development of AD. These findings may provide the therapeutic opportunities to AD.
Proteomic characterization of peripheral leukocytes in variant carriers of CDKAL1 associated with cholesterol efflux capacity
Authors: Lee S H , Ann S J , Choi D
Published: November 2023

Background

Genetic variations near CDKAL1 (CDK5 regulatory subunit-associated protein 1-like 1) gene recently showed association with individual’s cholesterol efflux capacity. The aim of this study was to compare the proteomes of peripheral leukocytes in CDKAL1 variant carriers and controls to identify evidence of biological impact caused by these variants.

Methods

Peripheral blood leukocytes were isolated from five individuals with any of four CDKAL1 variants (rs117835232, rs117252933, rs118064592, and rs150434350) and five controls. Cell lysates were digested to peptides and Q Exactive Orbitrap Plus mass spectrometry was applied for comparative proteomics analysis. Protein identification and quantification were performed with MaxQuant software.

Results

In total, 2,387 proteins were identified at false discovery rate 1% level, of which 176 proteins were differently abundant between CDKAL1 variant and control groups (p <0.05, fold-difference >1.5). Among them, 39 upregulated and 137 downregulated proteins were identified in the variant group (p <0.05; fold change>1.5) (Figure 1). The upregulated included docosahexaenoic acid omega hydroxylase, resistin, and apoA1, whereas the downregulated included NADH ubiquinone oxidoreductase. The functional enrichment and KEGG pathway analysis showed 20 associated pathways including carbon metabolism, synthesis of antibiotics, and RNA transport.

Conclusions

Differential expression of proteins regulating lipid metabolism and RNA transport in CDKAL1 variant carriers indicates differences in corresponding biological function in these individuals. Detailed contribution of the reported proteins remains to be clarified.
Cyclodextrin-loaded nanobubbles reduce cholesterol and atherosclerosis in vivo
Authors: Wang X , Mehta S , Bongcaron V , Nguyen T , Jirwankar Y , Maluenda A , Walsh A P G , Palasubramaniam J , Hulett M , Srivastava R , Bobik A , Peter K
Published: November 2023

Background

Ischemic heart disease (IHD) is the leading cause of mortality and morbidity worldwide. Atherosclerosis, a chronic inflammatory disease, is the main contributor to IHD. However, atherosclerosis is commonly left undiagnosed and untreated. Our study aims to create a theranostic nanobubbles (NBs), with ultrasonic properties for diagnostic imaging and at the same time for the delivery of 2-Hydroxypropyl-beta-cyclodextrin (CD), an FDA-approved drug to dissolve cholesterol crystals.

Methods and Results

Using polybutlycyanoacrylate, we created polymeric NBs. The incorporation of near-infrared dye (IR780) resulted in a multimodal contrast NB, which allows in vitro and in vivo visualization on both ultrasound and optical imaging. Furthermore, these NBs can be triggered by ultrasonic bursts to release their dye/drug payload. The safety profile of the NBs, post-incorporation of CD (CDNBs), demonstrated excellent biocompatibility in vitro using cell culture and in human blood. Cellular uptake of CP increased by 2.4x when using CPNBs compared to free CD (p<0.00001). In vivo biodistribution demonstrated that the NBs circulate in blood plasma for 48 hours, with a peak at 1-hour post-injection. In a mouse model of atherosclerosis, ApoE-/- mice on high-fat diet, we intravenously injected CDNBs, free CD or NBs only fortnightly for 8 weeks. En face Oil Red O-staining showed a significant reduction in total plaque area in the CDNB-treated group, as compared to the control animals treated with free CD and with NBs only (p>0.01). There was also a significant reduction in blood plasma cholesterol for the CDNBs-treated mice as compared to the control mice (p>0.01).

Conclusion

We demonstrated the ability to load CDs into NBs. These CDNBs are taken up by immune cells. The passive accumulation of CDNPs in the atherosclerotic lesions, resulted in the release of CD and the reduction of total plaque area in the entire aorta. These CDNBs treated mice also have a reduced cholesterol level in their plasma. CDNBs represent a novel theranostic nanocarrier with a high potential of clinical translation and ultimately major benefits to patients with atherosclerosis.
The effect of antibody-mediated PCSK9 inhibitor on bare-metal stent implantation outcome
Authors: Puspitasari Y M , Ministrini S , Liberale L , Vukolic A , Baumann-Zumstein P , Holy E W , Montecucco F , Luscher T F , Camici G G
Published: November 2023

Introduction

Percutaneous coronary intervention (PCI) is the gold standard for the management of acute coronary syndrome, and has been benefited from advances in pharmacotherapy and device innovation. Nevertheless, in-stent restenosis and stent thrombosis remain serious complications following PCI procedure with stent implantation. Proprotein convertase subtilisin kexin type 9 (PCSK9) is an enzyme known for its role in plasma cholesterol homeostasis. Efficiently reducing plasma cholesterol, PCSK9 inhibition recently became a pivotal treatment strategy for hypercholesterolemia. However, the effect of PCSK9 inhibition on stent implantation outcomes remains uninvestigated.

Methods

Carotid artery bare-metal stent (BMS) implantation was performed on C57Bl/6 mice, which then a parallel assessment of various inflammatory mediators was performed. To inhibit PCSK9, alirocumab was administered weekly to antiplatelet-treated mice started before stent implantation. Six weeks after the stent implantation procedure, animals were harvested and histomorphometric analysis was performed on stented arteries. In vitro cell migration and cellular senescence were also assessed on alirocumab-treated primary human aortic endothelial (EC) and vascular smooth muscle cells (VSMC).

Results

Compared to sham intervention, stent implantation was associated with increased expression of several inflammatory mediators, including PCSK9. Interestingly, the increase in PCSK9 level was confirmed in the stented vascular tissue, but not in plasma (Fig 1A). Following alirocumab treatment, PCSK9 inhibition results in an increased intimal hyperplasia in the stented vascular segment of alirocumab-treated animals compared to controls (Fig 1B). In vitro, alirocumab promoted migration and inhibited the onset of senescence in primary human VSMC, while blunted the migration and increased the senescence of EC.

Conclusion

The results of this study demonstrated that antibody-based PCSK9 inhibition worsen bare-metal stent implantation outcome in mice. This observed effect is mediated, in part, by the differential effect on VSMC and EC senescence, thus promoting in-stent intimal hyperplasia while blunting vascular healing. The herein-reported data warrant additional investigations concerning the use of PCSK9 inhibitors in patients undergoing PCI with stent implantation.
Phenylacetylglutamine causes a pathologic inflammation state and enhances atherosclerosis through the b2-adrenergic receptor cAMP PKA NF-kappaB pathway in diabetes
Authors: Huang L
Published: November 2023

Background

Type 2 diabetes (T2DM) affects billions of patients and has a pathophysiologically important inflammatory component. Macrophage plasticity in diabetes is impaired, and remains in a persistent proinflammatory state; yet currently, the cause of the inflammatory state is unknown. Phenylacetylglutamine (PAGln) is an amino acid metabolism end product that rises in T2DM patients and is associated with incident major adverse cardiovascular events. However, the role of PAGln in the inflammatory component of T2DM remains unknown.

Purpose

Here, we aim to identify blood-based systemic factors for pathologic inflammation status and risk of atherosclerosis as they are highly reproducible at low cost and search for effective and safe agents to attenuate diabetic inflammatory complications.

Methods

In vivo, phenylacetylglutamine was dissolved in normal saline and injected in mice daily (intraperitoneal). The levels of PAGln were analyzed by liquid chromatography-mass spectrometry. The effect of PAGln on atherosclerosis progression was analyzed by Ldlr–/– mice, followed by a Western diet feeding for 12 weeks. In vitro, primary mice bone marrow-derived macrophages (BMDMs) were treated with phenylacetylglutamine (100 uM) for the indicated time. Elisa, PCR, and western blot were used to characterize the impact of PAGln on BMDMs and mice arteries. Flow cytometry was taken to quantify and type immune cells in mice bone marrow, blood, spleens, and arteries. The activity of the NF-κB pathway was assessed by KEGG analysis, western blot, luciferase, and activity assay.

Results

In vivo, increased inflammatory macrophages were detected in mice blood, spleens, bone marrow, and arteries after PAGln injection. In vitro, PAGln leads to macrophage M1 polarization. Taken together, PAGln disturbed the transition of macrophage phenotype from inflammatory to reparative, facilitating pathologic inflammation. The RNA of BMDMs after PAGln treatment was collected for RNA-seq. KEGG analysis and western blot prove PAGln exerted its function through the activation of NF-κB. PAGln could promote NF-κB signals by increasing the phosphorylation level of p65. Besides, the negative effects of PAGln could be rescued by ß-blockers.

Conclusion

Our study unravels a novel mechanism by which PAGln accelerates atherosclerosis progression and causes pathologic inflammation by activating the ß2-adrenergic receptor /cAMP/PKA/NF-κB pathway. The negative effect of PAGln could be rescued by carvedilol (ß blocker). Thus, our recognition of chronic inflammation in diabetes needs to be revised, and understanding these mechanisms can inform alternative treatment strategies and refine atherosclerosis repair therapies for diabetic patients.
Synergistic effect of early rhythm control and healthy lifestyle on the risk of dementia in patients with new-onset atrial fibrillation
Authors: Kim Y , Lee S R , Choi E K , Lee S W , Han K D , Oh S , Lip G
Published: November 2023

Background

Early rhythm control (ERC) and healthy lifestyle (HLS) have been found to be related to lower dementia risk in patients with atrial fibrillation (AF), however, it has not been evaluated whether there is a synergistic effect between the two.

Methods

Using the Korean National Health Insurance Service database, patients who were newly diagnosed with AF between 2009 and 2016. Patients were categorized as follows: those who did not receive ERC including antiarrhythmic drug, cardioversion, AF catheter ablation within 2-year after AF diagnosis, and had 1 or more unhealthy lifestyle behaviors among current drinking, current smoking, and lack of regular exercise (group 1), those with HLS, but not receiving ERC (group 2), those receiving ERC, but with at least 1 unhealthy lifestyle behavior (group 3), and those with HLS and receiving ERC (group 4). To compare 4 groups, propensity score weighting method was used to balance the baseline characteristics of 4 groups. During follow-up, incident dementia, vascular dementia, and Alzheimer’s dementia were evaluated.

Results

A total of 199,947 patients were included. The mean time from AF diagnosis to implementation of ERC of groups 3 and 4 was 27±77 and 27±74 days, respectively. Among total study population, 96,907, 53,293, 29,234, and 20,513 patients were included in group 1, 2, 3, and 4, respectively. Group 4 showed the greatest risk reduction for all dementia, vascular dementia, and Alzheimer’s dementia among 4 groups (Figure). Both ERC alone and HLS alone were associated with a lower risk of vascular dementia by 20% than group 1. The combination of the two was associated with a lower risk of vascular dementia by 40% (Figure). However, the impact of ERC on the risk reduction of Alzheimer’s dementia was statistically insignificant, and maintaining HLS seemed sufficient to lower the risk of Alzheimer’s dementia (Figure).

Conclusion

The combination of ERC and HLS might be beneficial to prevent vascular dementia in patients with new-onset AF. To reduce the risk of Alzheimer’s dementia, maintaining HLS should be emphasized.
Vascular endothelial dysfunction induced by 3 bromofluoranthene via MAPK mediated NF B proinflammatory pathway and intracellular ROS generation
Authors: Su C H
Published: November 2023

3-Bromofluoranthene (3-BrFlu) is the secondary metabolite of fluoranthene, which is classified as a polycyclic aromatic hydrocarbon, through bromination and exists in the fine particulate matter of air pollutants. Endothelial dysfunction plays a critical role in the pathogenesis of cardiovascular and vascular diseases. Little is known about the molecular mechanism of 3-BrFlu on endothelial dysfunction in vivo and in vitro assay. In the present study, 3-BrFlu included concentration-dependent changes in ectopic angiogenesis of the sub-intestinal vein and dilation of the dorsal aorta in zebrafish. Disruption of vascular endothelial integrity and up-regulation of vascular endothelial permeability were also induced by 3-BrFlu in a concentration-dependent manner through pro-inflammatory responses in vascular endothelial cells, namely SVEC4-10 cells. Generation of pro-inflammatory mediator PGE2 was induced by 3-BrFlu through COX2 expression. Expression of COX2 and generation of pro-inflammatory cytokines, including TNFa and IL-6, were induced by 3-BrFlu through phosphorylation of NF-kB p65, which was mediated by phosphorylation of MAPK including p38 MAPK, ERK and JNK. Furthermore, generation of intracellular ROS was induced by 3-BrFlu, which is associated with the down-regulated activities of the antioxidant enzyme (AOE), including SOD and catalase. We also found that 3-BrFlu up-regulated expression of the AOE and HO-1 induced by 3-BrFlu through Nrf-2 expression. However, the 3-BrFlu-induced upregulation of AOE and HO-1 expression could not be revised the responses of vascular endothelial dysfunction. In conclusion, 3-BrFlu is a hazardous substance that results in vascular endothelial dysfunction through the MAPK-mediated-NFkB pro-inflammatory pathway and intracellular ROS generation.
PCSK9-inhibitor therapy leads to changes in immune cell gene expression and function: A subset analysis of the PACMAN-AMI trial
Authors: Ondracek A , Hofbauer T M , Aszlan A , Kuehn S , Seidl V , Toma A , Speidl W , Hengstenberg C , Raber L , Lang I M
Published: November 2023

Introduction

The PACMAN-AMI trial showed a significant reduction of plaque atheroma volume by alirocumab on top of high-intensity statin therapy as compared to placebo in 265 patients undergoing serial multi-vessel imaging after acute myocardial infarction (AMI). Neutrophils and the formation of neutrophil extracellular traps (NETs) have been implicated in the development and progression of atherosclerosis. High levels of low-density lipoprotein (LDL) have been associated with neutrophil activation.

Objectives

This study aimed to determine the influence of aggressive LDL-lowering on neutrophil activation, and immune cell gene expression of pro-inflammatory markers in a subset of the PACMAN-AMI cohort.

Methods

Neutrophils and peripheral blood mononuclear cells (PBMC) of 38 patients were isolated by serial density-gradient centrifugation to assess relative gene expression of IL-1b, IL-6, TNFa, X-box binding protein (XBP) 1, caspase 1 (CASP1), and NLRP3 by quantitative PCR. Neutrophils were stimulated to undergo NET formation using a calcium ionophore in the presence or absence of pro-protein convertase subtilisin/kexin type 9 (PCSK9), detecting the release of double-stranded (ds)DNA. Plasma samples were screened for dsDNA, citrullinated histone H3, neutrophil elastase, and myeloperoxidase. Healthy controls (n=30) were recruited during routine check-ups in a health and prevention center.

Results

After one year of double-blind injections, the probability to undergo NET formation in vitro was still higher in AMI patients than in healthy controls. Considering randomized treatment, only patients injected with alirocumab showed significantly higher NET release than controls. Additional stimulation with the alirocumab target PCSK9 could reduce NET formation in all cohorts. This effect was significantly higher in patients treated with alirocumab. Relative gene expression of CASP1, IL-1b, IL-6, and XBP 1 was higher in patients treated with alirocumab. There was no difference in plasma NET markers comparing healthy controls and both treatment arms.

Conclusion

Our data indicate that PCSK9 might have effects beyond its role in cholesterol metabolism influencing immune cell activation, which could mask effects of LDL lowering. Treatment does not directly translate to higher plasma NET levels without an additional inflammatory stimulus disrupting the steady state.
Development and internal-external validation of a prediction model for premature ventricular contraction unresponsive to the medical treatment
Authors: Atici A , Tanboga H İ , Barman H A , Sahin İ , Baycan Ö , Kup A , Celik M , Demirkiran A , Cevik E , Soysal A U , Karaduman M , Yilmaz İ , Yilmaz Y , Caliskan M , Aras D
Published: November 2023

Background

Ventricular premature contractions (PVCs) are the most common ventricular arrhythmias. However, to date, no risk stratification tool exists to assess response to medical therapy in patients with frequent PVCs (>5% per 24 hours). We aimed to develop and validate a clinical prediction model evaluating response to medical therapy in frequent PVCs.

Methods

We conducted a retrospective cohort study of patients with frequent PVCs who were considered for medical treatment. The study outcome was unresponsiveness to the medical treatment (24-hour PVC count not diminishing by at least 80%) in patients with frequent PVCs at 3-6 months follow-up. Potential predictors were, age (years), PVC burden (%), LVEF % (<55, ³55), PVC QRS width (msec), mean heart rate (beat/min), sinus beat QTc (msec), PVC coupling interval, gender, presence of multifocal PVC and Non-sustained VT. Binary logistic regression analyses were performed to develop and internally validate the model. Finally, we used internal-external cross validation using leave-one-out center.

Results

1644 patients were included in the study (mean age 52.2±13.5 years and 56.3% male). The frequency of unresponsiveness to the medical treatment in patients with frequent PVC was 31.2% (n=513). In the model, PVC burden (%), LVEF% (<55, ³55) and PVC QRS width were found to be the three strongest predictors. The apparent and internal validation discriminations of the model (C-statistics in internal validation 0.910) were quite satisfactory. Model discriminations and calibration metrics with internal-external cross validation were similar to the apparent model and were deemed acceptable (https://demonoreflow.shinyapps.io/dynnomapp/).

Conclusions

We developed and validated a model that accurately predicts unresponsiveness to medical treatment in patients with frequent PVC. Our model, once externally validated, has the potential to facilitate management decisions by providing individualized risk estimates in patients with frequent PVC for whom ablation may be suggested as a first line treatment modality.Figure-1Figure-2
Early ICD implantation and survival after out-of-hospital cardiac arrest
Authors: Myredal A , Rawshani A R A Z , Sultanian P E D R A M
Published: November 2023

Background

It is unclear whether an implantable cardioverter-defibrillator (ICD) is generally beneficial in survivors of out-of-hospital cardiac arrest (OHCA) of cardiac etiology.

Aim

We studied the association between ICD implantation and survival in patients with cardiac etiology or initial shockable rhythm in OHCA, using the Swedish Registry for Cardiopulmonary Resuscitation (SRCR).

Methods

All cases in the SRCR during 2010 to 2020 with cardiac etiology or initial shockable rhythm who were discharged alive included. We examined the association between ICD and survival using propensity score. Propensity scores (919 candidate predictors before feature selection) were estimated using gradient boosting, Bayesian additive regression trees, neural networks, extreme gradient boosting and logistic regression. Cox regression was used to model survival. The primary outcome was death or recurrent cardiac arrest.

Results

The 90-days event rate was 25% in the ICD group, compared with 52% in those without ICD. With regards to the average treatment effect (ATE) the overall hazard ratio 0.38 (95% CI 0.26-0.56) for ICD vs. no ICD. The overall average treatment effect on those treated (ATT) was 0.31 (95% CI 0.22-0.44). In the 15 subgroup analyses, every point estimate were below 1, with the most pronounced association noted in those with a history of acute myocardial infarction (HR 0.07 [95% CI 0.02-0.32]). All ATE and ATE estimates were aligned.

Conclusions

Mortality in survivors of OHCA occurs early after discharge, and prompt implantation of an ICD is associated with 62% lower probability of death and recurrent cardiac arrest overall.
Neurohormonal status is associated with long-term appropriate implantable cardioverter defibrillator therapy for life-threatening ventricular arrhythmia
Authors: Beygui F , Roule V , Guedeney P , Werner T , Zeitouni M , Ollitraut P , Silvain J , Alexandre J , Collet J P , Milliez P , Montalescot G
Published: November 2023

Background

Neurohormonal status is associated with the risk of life-threatening ventricular arrhythmia.

Purpose

We aimed to assess whether neurohormonal status in patients with an implantable cardioverter defibrillator (ICD) is correlated with the risk of long-term appropriate shock or overdrive delivered by the ICD for life-threatening arrhythmia.

Methods

A total of 331 clinically stable patients with ICD without any past history of shock or overdrive for life-threatening ventricular arrhythmia were enrolled in a two-center prospective cohort and followed up for a median time of 7.9 years. At enrolment, patients’ medical history, treatment, clinical and imaging data were recorded and, blood and, blood samples were drawn for the assessment of standard biological parameters as well as 3 neurohormones (aldosterone, A type NT-pro-Atrial natriuretic peptide (ANP) and MR-pro-Adrenomedullin (ADM).

The primary endpoint of the study was the occurrence of appropriate shock or overdrive delivered by the ICD (ICD therapy).

A multivariable cox model adjusted on age, gender, primary or secondary prevention ICD as well as all baseline variables unequally distributed between groups with or without ICD therapy was used for the assessment of the relationship between the 3 neurohormones and the primary endpoint.

Results

The population consisted of patients with a mean age of 60±15 years, among which 59(18%) women and 160(48%) with a secondary prevention indication for ICD. At baseline all data were similar between patients with or without ICD therapy with the exception of weight, systolic blood pressure, left ventricular ejection fraction (LVEF) and serum potassium and creatinine levels. No other variable, including MADIT and SHOCKED score were significantly associated with the risk of ICD therapy.

Death was recorded in 114(34%) patients and a total of 78(24%), 76(23%) and 106(33%) patients had appropriate shock, overdrive and ICD therapy respectively during the follow-up.

On multivariable analysis, highest levels of ANP and lowest levels of ADM but not aldosterone were independently associated with life-threatening ventricular arrhythmia requiring ICD therapy (table). Other correlates of ICD therapy were lower LVEF and, higher weight, serum potassium and creatinine concentrations. ANP and ADM had a stepwise additive impact on the risk of ICD therapy. (Figure 1)

Conclusions

In a stable all comer population with ICD and no history of ICD therapy, blood levels of neurohormones ANP and ADM are associated with the risk of life-threatening ventricular arrhythmia requiring ICD-delivered shock or overdrive independent of other correlates. ANP and ADM concentrations may be useful in identifying patients at high risk of such arrhythmia and who may most benefit from ICD implantation.
Mortality benefit of implantable cardioverter-defibrillators for primary prevention of sudden cardiac death: a real-world analysis
Authors: Auricchio A , Ahmed A , Mittal S , Pickett R A , Wilkoff B L , Jacobsen L , Marti A K , Soderland D M , Curtis A B
Published: November 2023

Background

Nearly 1,000 people suffer an out-of-hospital sudden cardiac arrest daily in the United States, with only 10% surviving when treated by emergency medical services. Implantable cardioverter-defibrillators (ICDs) are a proven first-line treatment in limiting sudden cardiac death, but the evidence has become dated. We sought to assess the survival benefit of ICDs in a contemporary population.

Purpose

Using a real-world database, we aimed to determine the current impact of ICDs on mortality in primary prevention patients.

Methods

Data were obtained from a de-identified electronic health record database. We included patients that met ICD primary prevention indication (PPI) implantation criteria between 2012 and 2019, had one year of available data before the PPI, and survived for at least one year post-PPI. Patients were split into 2 groups based on ICD implant status (implanted vs not implanted) within one year after PPI. The primary endpoint was the overall mortality rate, beginning at one-year post-PPI. Propensity matching was performed to account for baseline differences between patients that did and did not receive an ICD, and all-cause mortality rates were compared using the Kaplan-Meier method and Cox proportional hazard model.

Results

A total of 25,296 patients were identified as having a PPI for ICD implantation, of which 2,118 (8.4%) were implanted with an ICD within a year. Implanted patients were younger than non-implanted patients (63.4 vs 66.1 years) and had a smaller proportion of females (25.0% vs 36.7%). After 4 to 1 propensity matching, patients who did not have an ICD implanted had a mean age of 63.6 years and were 25.1% female. In the matched cohort, the 5-year all-cause mortality rate was 37.6% (CI: 33.2%-41.7%) in PPI patients who received an ICD and 44.9% (CI: 42.8%-47.0%) in those that did not receive an ICD. A Cox proportional hazard model revealed a 25.3% lower risk of all-cause mortality in PPI patients implanted with an ICD compared to PPI patients without an ICD (HR: 0.747; CI: 0.670-0.834; p<0.001 [Figure 1]).

Conclusion

In a large cohort of PPI patients that survived at least one year after indication, those implanted with an ICD during that year had a 25.3% lower risk of all-cause mortality than similar patients who did not receive an ICD, consistent with landmark trials. Thus, ICD implantation continues to be associated with a reduction in all-cause mortality in PPI patients at risk for sudden cardiac death.
Assessing the benefit of a replacement implantable cardioverter-defibrillator device for primary prevention of sudden cardiac death
Authors: Auricchio A , Curtis A B , Mittal S , Pickett R A , Jacobsen L , Lande J D , Wilkoff B L
Published: November 2023

Background

Implantable cardioverter-defibrillators (ICDs) have been shown to reduce the burden of sudden cardiac death in at-risk populations. However, data reflecting contemporary device programming and medication would be valuable, particularly when evaluating patients for a replacement ICD when they did not receive therapy from their original device.

Purpose

This real-world analysis examined the incidence of ICD therapy after routine generator replacement due to battery depletion in patients who didn’t receive therapy from their first ICD.

Methods

Data were obtained from a query of a deidentified database from 2007 to present. The final analysis included patients that underwent a generator replacement after 2012 due to battery depletion and who had received no therapy from their first ICD. Patients were divided into two groups based on their ICD indication status (primary prevention [PP] or secondary prevention [SP]). The main endpoint was time to first appropriate shock therapy, antitachycardia pacing (ATP) therapy, or any therapy (ATP or shock) after ICD replacement. Time to first therapy was plotted using the Kaplan-Meier method. Appropriateness of therapy was modeled via LASSO regression using characteristics present in both our dataset and the adjudicated PainFree SST trial. The model was fit on episodes from 80% of PainFree SST patients and validated on the remaining 20% before applying it to our dataset.

Results

A total of 8,383 patients were identified who met the criteria outlined above. Of these patients, 7,299 (66.3 ± 12.6 years old; 30.3% female) had a PP indication for an ICD and 1,084 (64.2 ± 14.0 years old; 30.3% female) had an SP indication. The mean longevity of initial ICD implants before replacement was similar in PP and SP patients (91.4 ± 16.2 vs 90.9 ± 16.3 months). At 7 years post-replacement, the incidence of first shock, ATP, or any therapy received by PP patients who did not receive first-device therapy was 16%, 23% and 27% respectively. After applying the model to estimate appropriateness of therapy, the appropriate therapy rates were 14%, 22%, and 26%, respectively (Figure 1). Similarly, SP patients who received no first-device therapy had an incidence of modeled first appropriate shock, ATP, and any therapy 7 years after ICD replacement of 12%, 20%, and 25%, respectively. The modeled 7-year inappropriate shock rate post-replacement was 4% in all patients.

Conclusion

In this large, long-term, real-world evaluation of ICD PP patients who hadn’t experienced a treated event with their initial device, 1 in 4 eventually received appropriate therapy from their second device. The results suggest that the underlying risk persists in these patients and device replacement may be necessary to protect them from experiencing a deadly event.
Comparison of two commercially available cryoballoon ablation systems
Authors: Diofano F , Pflieger C , Teumer Y , Bothner C , Rottbauer W , Dahme T , Weinmann K
Published: November 2023

Background

Cryoballoon pulmonary vein isolation (PVI) is an established anatomical single-shot procedure for the treatment of atrial fibrillation (AF).

Purpose

We compared efficiency of two commercially available cryoballoon ablation systems (CBS) to treat AF.

Methods

We prospectively enrolled 121 consecutive AF patients and treated them with either a compliant or a non-compliant CBS.

Results

Evaluation of procedural characteristics revealed a significantly longer procedure duration (p=0.007) and fluoroscopy time (p=0.008) in the group of the compliant CBS. The nadir cryoballoon temperature was lower using the compliant system compared to the non-compliant CBS (p<0.0001), likewise the time to isolation was not different between both groups. The non-compliant CBS showed a better occlusion for the right superior pulmonary vein (RSPV, p=0.039) leading to a higher rate of isolation with first freeze (p=0.004) and lower number of ablations at the RSPV (p=0.015).

A mean follow-up period of 310.8±189.7 days within a 90-day blanking period. 68% of patients treated with the non-compliant and 65% of patients treated with the compliant CBS showed freedom from atrial arrhythmia recurrence (p=0.724). Kaplan-Meier estimation showed no significant difference between both groups (LogRank p=0.768).

Conclusion

The novel compliant CBS showed a longer procedure duration and fluoroscopy time, as well as, a lower nadir cryoballoon temperature. The non-compliant CBS was more effective in isolation of the RSPV. Acute PVI success and long-term freedom from atrial arrhythmia recurrence after a mean follow-up of ten months did not differ between groups.