bluuebunny/crossref_metadata_2025 huggingface dataset:
Utility of Soluble ST2 biomarker to predict recurrence after electrical cardioversion in patients with atrial fibrillation
Authors: Gonzalez-Melchor L , Garcia-Seara J , Martinez-Sande J L , Rodriguez-Manero M , Fernandez-Lopez X A , Alvarez Villamarin C , Gonzalez-Juanatey J R , Minguito-Carazo C , Gonzalez T
Published: October 2022

Background

Atrial fibrillation (AF) is the more frequent arrhythmia in clinical practice. The ST2S is a biomarker that has demonstrated to be predictor of cardiovascular outcomes in patients with heart failure but there is scarce information of his utility in patients with AF.

Purpose

Considering the characteristics of the ST2S we hypothesize this biomarker could correlate to recurrence in patients with AF and electrical cardioversion (ECV).

Methods

This was an observational and prospective clinical trial. We compared all patients with AF referred for ECV with a control group without AF, from September 1th 2016 to September 30 2019. Clinical, ECG, echocardiographic and ST2S levels were analyzed in both groups at basal, at 3 and 6 months of follow-up in such cases with AF. Patients with inflammatory or allergic diseases, moderate/severe ventricular dysfunction, structural cardiomyopathy, moderate/severe hepatic, renal or respiratory disease were excluded.

Results

We included a total of 94 patients with AF and 40 paired controls. Clinical variables are presented in Table 1. Fifty-eight (61.7%) patients with AF had recurrence at follow-up. There was a significant difference between the ST2S levels at baseline between AF patients (17163.8 pg/mL) and controls (11016.2 pg/mL) (p=0.001). ST2S biomarker levels at 3 and 6 months of follow-up decreased in those patients without AF recurrence as shown in Figure 1. Covariable models were performed and ST2S biomarker levels at 3 months were significant to predict recurrence at 6 months follow-up (Table 2). The calculated cut-point of the biomarker was of 15511.51 pg/ml with a c-value: 0.669.

Conclusions

In our experience ST2S was a useful biomarker to predict recurrence of AF after ECV. Considering the size of the study more studies should be performed to confirm this results.

Funding Acknowledgement

Type of funding sources: None.
Cardiac troponin T degradation in end-stage renal disease patients appears to occur in vivo
Authors: Denessen E , Vroemen W H M , Litjens E J R , Henskens Y M C , Van Der Sande F M , Kooman J P , Bekers O , De Boer D , Mingels A M A
Published: October 2022

Background

Cardiac troponin T (cTnT) is a cornerstone biomarker for diagnosing myocardial infarction (MI). However, since the introduction of the high-sensitivity assays, cardiac troponins are also measured in pathologies with cardiac risk factors, such as end-stage renal disease (ESRD), making it more difficult for clinicians to diagnose MI. Previous studies of our group identified specific larger cTnT proteoforms for the acute phase of MI, while a study in serum of ESRD patients showed solely small cTnT fragments. However, others allocated the small cTnT fragments in serum of ESRD patients to a pre-analytic effect due to abundant thrombin generation in serum, as thrombin is known to cause fragmentation of cTnT.

Purpose

This study investigated the effect of multiple anticoagulation methods on cTnT proteoforms in vitro and in the blood of ESRD patients.

Methods

First, cTnT negative serum and lithium-heparinized (LH), ethylenediaminetetraacetic-acid (EDTA), and hirudin plasma tubes were spiked with human cTnT standard, incubated at 37°C till 48 hours, and analysed with Western Blotting using anti-cTnT Roche antibodies. Second, similar blood tubes were collected from ESRD patients (n=10) conform the Declaration of Helsinki, patient samples and spiked cTnT standards were separated using gel filtration chromatography (GFC) and cTnT was subsequently analysed using the Cobas 6000 analyser.

Results

After 48 hours of in vitro incubation of cTnT in hirudin plasma, no cTnT degradation was observed, while LH only showed minor degradation of 10% from intact 40 kDa cTnT to 29 kDa fragments (figure 1). Also, for both EDTA and serum, a time-dependent degradation from 40 kDa cTnT to 29 kDa fragments to 15–18 kDa fragments was observed. Moreover, in ESRD patients, GFC elution profiles of all blood tubes revealed that 85–98% of cTnT corresponded to small 15–18 kDa fragments (41–42 mL), while 2–15% were 29/40 kDa cTnT forms (27–28 mL). For comparison, standards of ternary T-I-C complex and intact cTnT eluted at 21 mL and 28 mL, respectively.

Conclusions

The extent of cTnT degradation in vitro is dependent on the anticoagulation method of the blood tubes, with absence of degradation exclusively in hirudin plasma. Since in ESRD patients for all blood tubes mainly small 15–18 kDa cTnT fragments were found, including hirudin plasma, cTnT degradation appears to occur in vivo. This observation supports the hypothesis that larger cTnT proteoforms might indeed be specific for the acute phase of MI, thereby supporting the opportunity for developing a more specific cTnT method.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): ZonMw Veni grant
Expression changes of circulating ACE2 regulating-microRNA profiles in patients with COVID-19 during hospitalisation
Authors: Jarosz-Popek J , Eyileten C , Wicik Z , Nowak A , Wolska M , Shahzadi A , Jakubik D , Simoes S N , Martins D C , Siller-Matula J , Postula M
Published: October 2022

Background

Various miRNAs, as well as their target genes are involved in the complex pathophysiology of inflammation, immune response, coagulation and CVDs. Thus, they may be useful for diagnosis, prognosis and as a potential therapeutic strategy in multiple pathologies.

Aim

According to our previously published bioinformatics analysis, we aimed to analyze the diagnostic and predictive utility of miRNAs regulating ACE2 network (miR-26b-5p, miR-10b-5p, miR-302c-5p, hsa-miR-200b-3p, hsa-miR-124-3p) in patients with COVID-19.

Results

The expression levels of miR-26b-5p in COVID-19 patients were found lower at the baseline, 7 and 21-days after admission. compared to the healthy controls (p<0.0001 for all time points). Similarly, miR-10b-5p expression levels were lower at the baseline and 21-days post admission in COVID-19 patients when compared to the healthy individuals (p=0.001 in both time points). Moreover, expression levels of this miRNA were higher 7-days post-admission when compared to the baseline (p=0.003). According to the ROC curve analysis, low miR-200b-3p expression presents predictive utility in assessment of the hospital length of stay and/or death (AUC: 0.730, p=0.002). According to the multivariable logistic regression model, low delta miR-200p expression, together with diabetes mellitus (DM), are independent predictors of increased hospital length of stay and/or death (OR: 5.775; 95% CI, 1.572–21.214; p=0.008 and OR: 4.888; 95% CI, 1.001–23.858; p=0.050, respectively).

Conclusions

This study enabled us to better characterize changes in gene expression and signaling pathways related to COVID-19 thrombosis. In this study we identified, characterized and validated miRNAs which could serve as novel, thrombosis-related biomarkers of the COVID-19, can be used for early stratification of patients and prediction of severity of infection development in an individual.

Funding Acknowledgement

Type of funding sources: Public Institution(s). Main funding source(s): Medical University of Warsaw
Proteomic analysis of exercise-induced hypertrophy reveals sex-related mitochondrial differences mediated by AMPK
Authors: Olah A , Barta B A , Ruppert M , Sayour A A , Bottlik O , Merkely B , Schilling O , Radovits T
Published: October 2022

Background

Regular physical activity results in characteristic structural and functional changes in the heart, which are collectively referred to as the athlete's heart. However, the extent of exercise-induced left ventricular (LV) hypertrophy and functional changes show significant differences between men and women, the molecular background of which is not fully elucidated.

Objective

The aim of this study was to provide a proteomic characterization of long-term, intense exercise-induced LV myocardial hypertrophy in a rat model, with a focus on sex-related differences.

Methods

Our rats were divided into trained (FEx) and control female (FCo) as well as trained (MEx) and control male (MCo) groups. In the trained groups, athlete's heart was induced by a 12-week swimming protocol. Myocardial hypertrophy was confirmed by echocardiography and functional adaptation by pressure-volume analysis. Proteomic measurements based on liquid chromatograph-coupled mass spectrometry were performed on proteins isolated from our LV myocardial samples.

Results

Echocardiography and post-mortem myocardial mass showed significant LV hypertrophy in both sexes, which was more pronounced in female animals (tibial length normalized LV muscle mass: + 17.4% MEx vs. MCo, + 31.0% FEx vs. FCo). LV contractility increased to the same extent in both sexes. Relative expression of 3074 proteins were determined by proteomics. There was a significant change in expression of 229 proteins in males and 599 in females compared to the level of same-sex controls. Based on our gene ontological analysis, physiological LV remodeling in females is characterized by increased expression of proteins in mitochondrial function (cellular respiration and fatty acid oxidation) and biogenesis, whereas in males, proteins that bind to the actin cytoskeleton is primarily increased. Further investigation revealed that the quantity of AMP-activated protein kinase (AMPK) and sirtuin 3 (SIRT3) was increased only in female animals.

Conclusions

Our data suggests that physiological LV hypertrophy resulting from regular, balanced exercise is associated with sex-specific changes in the myocardial proteome. The main differences might be associated with different regulation of mitochondrial function and biogenesis, related to AMPK pathway. Our results contribute to the understanding of the development of physiological myocardial hypertrophy.

Funding Acknowledgement

Type of funding sources: Private grant(s) and/or Sponsorship. Main funding source(s): Bolyai János Research Scholarship (BO/00837/21) to OANational Research, Development and Innovation Office (NKFIH) K135076 to B.M.
A promising rat model of heart failure with preserved ejection fraction associated to multiple comorbidities
Authors: Dubroca C , Costard C , Maupoint J , Waget A , Brousseau E , Briand F , Sulpice T
Published: October 2022

Background and aims

Heart failure with preserved ejection fraction (HFpEF) is a complex clinical phenotype often associated to the presence of multiple non-cardiac interrelated comorbidities including diabetes, obesity, hypertension. A major challenge in the development of new therapies for HFpEF is the lack of animal models that truly recapitulate the complexities of the disease. One promising candidate is the Lund MetS rat, a congenic BBDR.cg-lepr.cp model generated by introgression of the Koletsky leptin receptor mutation into the BioBreeding Diabetes Resistant (BBDR) rat.

Methods

14-week-old Lund MetS obese T2D rats were exposed to salt enriched diet (0.8% NaCl in diet) for 6 weeks. Age-matched lean rats (ctrl) were fed a normal chow diet. Cardiovascular phenotyping was assessed by echocardiography and blood pressure measurement. Vascular function of isolated thoracic aortas, as well as kidney and cardiac parameters were also evaluated at the end of the 6-week diet period.

Results

After 6-week enriched salt diet period, mean arterial pressure increased from 120mmHg to 160mmHg (vs 100mmHg in chow-fed lean rat). The hypertensive state also led to renal hypertrophy with up to a 2-fold higher kidney weights. Echocardiography showed a preserved systolic function with quite similar ejection fraction and fractional shortening in Lund MetS and lean rats. In contrast, high salt-fed Lund MetS rats developed cardiac hypertrophy confirmed by a large increase in heart, left ventricle, and left atria weights as compared to lean rats. Diastolic dysfunction was also highlighted with inverted E/A and E'/A' ratios along with a prolongation of the isovolumic relaxation time (IVRT). This was accompanied by an increase in circulating levels of soluble ST2 and NT- proBNP. Finally, aortic rings assays showed that both endothelium dependent and independent vasodilating properties were impaired in the high salt-fed Lund MetS rats.

Conclusions

The present data demonstrate that the Lund MetS rat fed a high salt diet developed a marked hypertension, pathological cardiac remodeling and diastolic dysfunction without modification of ejection fraction, along with obesity and type 2 diabetes. Thus, this could be a relevant model to bring forward effective new treatments for HFpEF patients.

Funding Acknowledgement

Type of funding sources: None.
Alterations in the gut microbiota composition in the obesity phenotype of heart failure with preserved ejection fraction
Authors: Rokach Y , Abedat S , Nachman D , Dana S , Qadan A , Beeri R , Amir O , Asleh R
Published: October 2022

Introduction

Obesity is one of the most common phenotypes of heart failure with preserved ejection fraction (HFpEF). A growing body of evidence indicates that obesity is characterized by compositional alterations in the intestinal microbiota and its metabolites, collectively referred as dysbiosis.

Purpose

We sought to investigate whether the obese phenotype of HFpEF is mediated by further alterations in the microbiota and whether gut dysbiosis fuels metabolic inflammation leading to progression of HFpEF.

Methods

HFpEF was induced in C57/black mice by high-fat diet and inhibition of NO synthesis (a “2-hit-model”) (Figure 1A). High-frequency echocardiography was performed to assess diastolic function. Blood pressure (BP) was measured using CODA non-invasive tail-cuff system. For human studies, patients with obese HFpEF as well as obese and non-obese healthy individuals were prospectively recruited. Gut microbial DNA was subjected to 16S ribosomal RNA gene sequencing, and the differences in abundance and composition of bacteria were defined both in humans and mice.

Results

In mice, the 2-hit model demonstrated significantly increased BP and impaired diastolic function in the obese HFpEF group (Figure 1B). Microbiome analysis showed significant differences in the gut microbial diversity and composition in the obese HFpEF compared to the non-HFpEF obese and chow-fed mice (Figure 1C). Similarly, human microbiome sequencing showed significant gut bacterial alterations in the diversity and composition of bacteria with more profound dysbiosis observed in the obese HFpEF patients compared to the obese and lean healthy individuals (Figure 1D).

Conclusions

We provide evidence in humans as well as in a murine animal model that the obese HFpEF phenotype is associated with more pronounced gut dysbiosis compared to the non-HFpEF obesity state. Further research will elucidate the causative link between gut dysbiosis and HFpEF manifestation and the mechanistic insights into the implication of gut microbiota in HFpEF progression.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): Israel science foundation
Acute afterload leads to increased electrophysiological heterogeneity after myocardial infarction
Authors: Ingelaere S , Vermoortele D , Holemans P , Claus P , Willems R
Published: October 2022

Background

Myocardial infarction (MI) results in altered mechanical loading and changes in the cardiac electrical properties. The infarct border zone is pro-arrhythmic but the exact role of mechano-electrical coupling remains unclear.

Objective

We studied spatial electrical heterogeneity in MI animals during acute afterload increase using a novel E-field methodology for high resolution mapping of local activation-repolarization intervals (ARI) in vivo.

Methods

Anterior-septal MI was induced in five domestic pigs by 120-minute occlusion of the left anterior descending artery followed by reperfusion. This led to an infarct size of 17.7±2.1% of the left ventricle. After 1 month, electro-anatomical mapping was performed before and during an acute afterload challenge induced by partially inflating a balloon in the descending aorta. A non-contact recording of a 64-electrode array was translated to 2048 non-contact electrograms distributed over the left ventricle. The non-contact electrograms were processed to determine the ARIs using a custom-made algorithm, previously validated against monophasic action potential recordings. Based on the contact map we defined border zone (BZ, voltage 0.5 to 1.5 mV) and remote (>1.5mV) regions. Heterogeneity was defined as the interquartile range (IQR) of ARIs in fixed neighborhoods of 1cm radius (figure 1A) and analyzed in 10 segments (5 BZ and 5 remote) of a modified version of the AHA model (49 segments by dividing the 16 non-apical segments). Other segments were discarded due to artefacts mainly caused by the array touching the septal and apical wall.

Results

Acute afterload challenge resulted in an increase of the systolic left ventricular pressure of 41.7±5.4% and increased left ventricular repolarization heterogeneity (IQR 4.03±1.23ms baseline to 4.85±1.38ms during inflation, p=0.004). There was a significant increase in heterogeneity in both BZ (4.78±1.60ms to 5.64±1.66ms, p=0.020) and remote (2.24±0.17ms to 3.00±0.86ms, p=0.034) regions (figure 1B). The IQR in the infarct BZ was higher compared to the remote zone at rest (4.78±1.60ms vs 2.24±0.17ms, p=0.010) as well as during inflation (5.64±1.66ms vs 3.00±0.86ms, p=0.008) (figure 1B). Both BZ and remote regions responded equally to acute afterload (p for interaction = 0.803).

Conclusion

Increased afterload leads to increased repolarization heterogeneity. This heterogeneity is higher in the infarct BZ. These alterations could provide a functional substrate for reentry.

Funding Acknowledgement

Type of funding sources: Public Institution(s). Main funding source(s): KU Leuven - C1 funding
Real-time electrophysiological characterization of acute and chronic radiofrequency ablation lesions
Authors: Amoros-Figueras G , Casabella-Ramon S , Moreno-Weidmann Z , Company-Ramon G , Jorge E , Rosell-Ferrer J , Cinca J , Guerra J M
Published: October 2022

Background

Assessment of lesion formation remains one of the most important goals to guide radiofrequency (RF) procedures in the cardiac electrophysiology laboratory. Cardiac navigation systems use specific algorithms to predict the effective lesion size, but these metrics are based only on ablation parameters and ignore local electric tissue characteristics. [1] Recent studies show that local multiparametric impedance is influenced by the intrinsic structural characteristics of the tissue. [2]

Purpose

This study aimed to assess the ability of local multiparametric impedance to characterize acute and chronic RF ablation lesions in the right atrium of pigs.

Methods

Four anesthetized closed-chest pigs were submitted to two interventions. The first intervention aimed at creating RF ablation lesions (30W, 60s, 55°) in the right atrium using an electrocatheter connected to a conventional cardiac navigation system, while mapping the local multiparametric impedance (LMI), the generator impedance (GI) and the bipolar voltage (BiV). All RF ablation lesions were performed with a stable contact force. Four weeks later, the second intervention aimed at mapping again the previous ablation sites. After the second intervention animals were euthanized and the hearts were removed and processed to identify the presence of fibrosis in the previously ablated lesions (Figure).

Results

We performed 19 ablations (5±2 per animal) with an average contact force of 14.2±4.6g. Four weeks later, 14/19 (74%) ablation lesions were identified as fibrotic points in the explanted heart (Figure 1). These showed decreased acute LMI and BiV values that persisted low after 4 weeks (Table 1). The remaining 5/19 ablations that did not present fibrosis had lower baseline LMI and bipolar values that resulted in lower LMI and BiV drops (Table 1). The absolute drop in LMI between effective and non-effective lesions was around 50%, while GI drop was only 21% (LMI drop @51KHz: From −3.0±1.4° to −1.5±0.3°, T-TEST p<0.05; GI drop: From 12.7±27.6Ω to 10.5±21.3Ω, T-TEST p=0.89).

Conclusion

The local baseline electrical properties of atrial myocardium are directly related to the effectiveness of RF ablation lesions. RF ablation sites that resulted into persistent fibrosis points had larger LMI and BiV values, with higher absolute drops than non-effective lesions. The use of LMI in clinical practice could improve the outcome of the procedures of arrhythmia ablation in the electrophysiology lab.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Supported by grants from ISCI-MINECO (FIS PI21/00392), FEDER, CIBERCV (CB16/11/00276)
Empagliflozin attenuates arrhythmogenesis via inhibition of O-GlcNAcylation in diastolic phase of diabetic cardiomyopathy
Authors: Kadosaka T , Watanabe M , Natsui H , Koizumi T , Koya T , Nakao M , Hagiwara H , Kamada R , Temma T , Anzai T
Published: October 2022

Background

Diabetic cardiomyopathy is an important complication of diabetes mellitus (DM) and reported to increase the risk of fatal ventricular arrhythmias. Recent clinical trials showed that empagliflozin (EMPA), a selective sodium-glucose co-transporter 2 (SGLT2) inhibitor, improved cardiovascular outcomes regardless of the presence of diabetes and the traditional cardiovascular risk factors. Despite the promising benefit of EMPA on heart failure treatment, its beneficial effect in the context of anti-arrhythmic therapy has not been fully examined. We therefore aimed to examine anti-arrhythmic effect of acute EMPA treatment especially onto calcium (Ca2+) handling in diabetic cardiomyocytes.

Methods

We assessed echocardiography, hemodynamic study, electrophysiology, Ca2+ handling and protein expression in C57BLKS/J-leprdb/db mice (db/db mice), a leptin receptor-deficient model of obesity and Type 2 diabetes, and their non-diabetic lean heterozygous Leprdb/+ littermates (db/+ mice).

Results

The 16-week-old db/db mice had preserved systolic function but exhibited diastolic dysfunction. In arrhythmia induction using ex-vivo Langendorff-perfused hearts, db/db mice showed a significantly increased premature ventricular complex (PVC) by 2x Ca2+ and 1μM isoproterenol load than control, which was attenuated by EMPA perfusion (1 μM). Diabetic cardiomyocytes showed an increased frequency of spontaneous Ca2+ sparks and waves, and decreased Ca2+ transient amplitude and sarcoplasmic reticulum (SR) Ca2+ content. Ca2+ transient decay tau and time to 50% decay were significantly prolonged in diabetic cardiomyocytes. These data indicating the impaired Ca2+ handling in diabetic cardiomyocytes were normalized by acute administration of EMPA (1 μM), while the administration of NHE inhibitor (Cariporide 10 μM) did not show significant differences. In the protein expression analysis, CaMKII Thr287 autophosphorylation and CaMKII-dependent RyR2 S2814 phosphorylation were significantly increased in diabetic hearts, which were inhibited by short-term (30 min) perfusion of EMPA (1 μM). The expressions of SERCA2a and phospholamban were not significantly different among three groups. Lastly, whole hearts O-GlcNAcylation, one of the important post translational modifications, was significantly reduced by EMPA treatment.

Conclusion

EMPA improved intracellular Ca2+ handling and attenuated arrhythmogenesis in mice with diabetic cardiomyopathy at the diastolic dysfunction phase, suggesting that EMPA may exhibit this anti-arrhythmic effect by normalization of intracellular calcium handling via inhibiting O-GlcNAcylation.

Funding Acknowledgement

Type of funding sources: Private grant(s) and/or Sponsorship. Main funding source(s): Boehringer Ingelheim
Spontaneous coronary artery dissection: current evidence from cardiac magnetic resonance and angiography screening
Authors: Androulakis E , Azzu A , Papagkikas P , Antonopoulos A , Al-Hussaini A , Pennell D , Mohiaddin R
Published: October 2022

Purpose

To assess the distribution and extent of myocardial infarction, as well as possible clinical significance of peripheral arteriopathy screening in a cohort of patients presented with spontaneous coronary dissection (SCAD) using cardiovascular magnetic resonance (CMR) for cardiac and vascular imaging. The adverse CMR features and predictors of infarction in subgroup analysis were also investigated.

Methods

This is an observational, single centre study of 144 consecutive, angiographically confirmed SCAD survivors >18 years old, based on collected data from Jan 2008-Nov 2020. All scans were performed on a 1.5-Tesla scanner. Cardiac structural and functional indices, myocardial infarct size and distribution were evaluated. In addition, vascular imaging was performed in all patients for thoracic aorta screening as part of the routine protocol, and in 75 patients peripheral arteriopathy screening from Circle of Willis to the iliac arteries was performed using 2D multi-slice and cine imaging, as well as 3D imaging methods.

Results

In the total population, 64% had infarction. Infarct size (%) was predictive of reduced left ventricular ejection fraction (LVEF) (p<0.001) and increased diastolic volumes (p<0.001 for both). Logistic regression showed SCAD in right coronary artery (Odds Ratio (OR): 5.2, p=0.034) predicted the presence of infarction. In the group who also underwent peripheral vascular screening, the overall prevalence of extra-coronary arteriopathy was 27% with fibromuscular dysplasia accounting for one-fourth of the cases (Figure 1). Of note, most positive vascular screening patients (73.3%) exhibited infarction (OR: 7.0, p=0.041), distributed in more than one territory (OR: 4.0, p=0.015) (Figure 2), while infarct size was negatively correlated with LVEF (p<0.001). Right ventricular ejection fraction was significantly reduced in positive screening patients (58.8±7.4 vs. 63.0±8.0, p=0.045). Further subgroup analysis showed pregnancy associated SCAD patients had significantly larger sized infarcts (25.1±6.1 vs. 6.2±0.7%, p<0.001) with reduced LVEF (51.0 vs 63.6±7.6, p<0.001). In logistic regression, this condition also predicted large infarcts >10% (OR: 9.8, p=0.009) in anterior walls (p<0.001).

Conclusions

Patients with SCAD require careful assessment of myocardial structure, function and characterization of infarction which may reflect the underlying pathophysiology. Extra coronary vascular screening, although not established in clinical practice, seems a rational strategy to potentially identify a subgroup with more severe infarction and likely to have vascular pathology.

Funding Acknowledgement

Type of funding sources: None.
Aberrant interaction between TEAD1 and Lamin A/C causes cardiomyopathy
Authors: Yamada S , Ko T , Ito M , Sassa T , Nomura S , Komuro I
Published: October 2022

Background

Mutations in the LMNA gene encoding Lamin A/C, a major component of the nuclear lamina, cause laminopathies including dilated cardiomyopathy (DCM). DCM patients with LMNA mutations have particularly severe clinical courses such as heart transplantation and death due to heart failure. However, underlying mechanisms of LMNA-induced DCM remains elusive.

Methods and results

We identified LMNA Q353R mutation in a DCM family with severe heart failure. We generated Q353R heterozygous knock-in mice, which showed sarcomere dysplasia and perinatal lethality. Integrative single-cell analyses of the fetal murine hearts and patient-specific induced pluripotent stem cell-derived cardiomyocytes (iPSCMs) revealed that transcriptional regulation of cardiomyocyte maturation/development genes governed by TEAD1 was impaired in LMNA mutant cardiomyocytes. Protein array and immunostaining uncovered increased binding of TEAD1 to mutant Lamin A/C protein and abnormal localization of TEAD1 at the nuclear periphery. Furthermore, TT-10, a Hippo pathway inhibitor, rescued the dysregulation of cardiac developmental genes in LMNA mutant cardiomyocytes. Single-cell RNA-seq of cardiac tissues from DCM patients with the LMNA Q353R mutation confirmed the dysregulated expression of TEAD1 and its target genes. These results demonstrated abnormal interaction between TEAD1 and mutant Lamin A/C impairs structural maturation of cardiomyocytes and suggests that LMNA Q353R-related DCM can be treated through intervention in the Hippo pathway.

Conclusion

TEAD1 trapped by mutant Lamin A/C protein at the nuclear membrane perturbs transcriptional maturation in LMNA Q353R-related DCM.

Funding Acknowledgement

Type of funding sources: None.
Characterization of myocardial proteomics in biopsy proven cardiomyopathies
Authors: Lee S H , Chang D K , Kyoung Y J , Kim E S , Kim I C , Youn J C , Kim J S
Published: October 2022

Background

Difference in proteomic expression according to the etiology of cardiomyopathies is not well known.

Purpose

We aimed to identify proteome based pathogenesis in patients with biopsy proven cardiomyopathies.

Methods

Comparative proteomic analysis of biopsy specimens were performed from 9 patients with cardiomyopathy (3 dilated cardiomyopathy (DCM), 2 hypertrophic cardiomyopathy (HCM) and 4 myocarditis) as well as 5 controls (normal endomyocardial biopsy specimen from one-year surveillance of heart transplant recipients) by tandem mass tag combined with liquid chromatography-mass spectrometry. Differential expression protein analysis, gene ontology analysis and Ingenuity pathway analysis were done to discover molecular mechanism for the differentially expressed proteins in each cardiomyopathy compared to the control.

Results

Differential expression protein analysis showed higher proportion of significantly increased proteins (Log2 fold change ≥1) in HCM and myocarditis, whereas higher proportion of significantly decreased proteins (Log2 fold change ≤−1) in DCM compared to controls (Figure). According to the gene ontology analysis, upregulation of neutrophil degranulation, and down-regulation of mitochondrial translation protein was noted in patients with DCM. In patients with HCM, platelet degranulation protein was increased, and mitochondrial ATP synthesis coupled electron transport, was decreased. In patients with myocarditis, neutrophil related proteins and calcium ion binding protein was increased, but muscle cell development, protein was decreased. Ingenuity pathway analysis revealed downregulation of oxidative phosphorylation and upregulation of sirtuin signalling pathway both in DCM and HCM. In myocarditis, various pathways related to inflammation were upregulated with only RHO GDP dissociation inhibitors downregulated.

Conclusions

This study showed that each cardiomyopathy exhibited different proteomic expression compared to normal heart. Further large detailed study is needed to understand the association between proteomic expression and disease pathophysiology.

Funding Acknowledgement

Type of funding sources: None.
Loss of ceramide synthase 5 reduces the development of aortic valve stenosis in mice with high fat diet
Authors: Reese L , Haenschke L , Niepmann S T , Bulic M , Duesing P , Zimmer S , Nickenig G , Bauer R , Jansen F , Zietzer A
Published: October 2022

Background

Ceramide synthase 5 (CerS5) is essential to maintain C:16 ceramide levels in mice. Ceramides contribute to inflammation and calcification, two hallmarks of aortic valve stenosis development. It is known, that loss of CerS5 function reduces diet-induced obesity.

Purpose

The aim of this study was to investigate the effect of ceramide synthase 5 and high-fat diet on the development of aortic valve stenosis with a mouse model.

Methods

Our experiment consists of four groups of mice: Wild type (WT) and CerS5 (−/−) mice, with normal and high fat diet. We induced the aortic valve stenosis development through a wire-injury. For this model, a coronary angiography wire is introduced into the left ventricle over the right carotid artery, followed by rotation on the aortic valve level to generate a defined injury. Development of aortic valve stenosis was determined by measuring the peak velocity 14, 28 and 42 days after the wire-injury. With histological analysis of the aortic valve, we measured macrophage infiltration with CD68 immunostaining and calcification using von Kossa staining. For statistical analysis ANOVA and Turkeys multiple comparisons were performed.

Results

Histological analysis of the aortic valve revealed, that in CerS5 (−/−) mice with high fat diet (HFD) immune cell infiltration is reduced, while there is no difference between CerS5 (−/−) mice and wild type, when fed with normal diet (A). Also calcification showed a trend towards a reduction in CerS5 mice with high fat diet, but was not statistically significant (B). Echocardiography could detect a reduced peak velocity of CerS5 mice with high fat diet, in comparison to the other groups, four and six weeks after wire injury (C).

Conclusion

Our experiments indicate that loss of CerS5 function reduces the development of aortic valve stenosis in mice with high fat diet, by decreasing immune cell infiltration and calcification. Interestingly, in the groups with normal chow, loss of CerS5 function had no effect. In addition, high-fat diet alone had no negative effect in our mouse model. Based on our results it can be assumed that upon high fat diet CerS5 mediates for pro-inflammatory effects in the aortic valve, which are absent on normal diet.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): Ernst & Berta Grimmke Foundation (13/19)
SGLT1/2 expression in the heart of transplanted patients is associated with inflammatory induction of oxidative stress: potential protective effect of SGLT2 inhibitors
Authors: Gong D S , Mroueh A , Matsushita K , Fakih W , Auger C , Oak M H , Pieper M P , Morel O , Epailly E , Schini-Kerth V
Published: October 2022

Introduction

Heart transplantation is a life-saving surgical procedure for patients with end-stage cardiac dysfunction, however, such procedures are usually at risk of rejection due to acute inflammatory activation. While such inflammatory induction leaves the newly transplanted heart at risk of functional and structural remodeling subsequent to oxidative damage, current anti-inflammatory treatment options expose the patient to an elevated risk for adverse reactions in addition to absence of cardio-protective effects. Therefore, novel therapies with anti-inflammatory and cardio-protective dual effects are needed. Recently, our group has reported that low-grade inflammation is associated with upregulation of SGLT1/2 in arteries of human with cardiovascular diseases. Yet, the role and function of SGLT2 in human cardiac tissue remains poorly understood.

Aim

This study focuses on the expression pattern of SGLT1/2 in cardiac biopsies of heart transplanted patients and aim to identifying their functional impact.

Methods

Routine endomyocardial Biopsies (23) were performed for the detection of acute rejection heart transplanted patients (less than 2 years) at our University Hospital. Gene expression levels were assessed using RT-qPCR, the in situ tissue localization of proteins by immunofluorescence staining, and the level of oxidative stress by dihydroethidium staining.

Results

Gene expression analysis revealed strong inflammatory reaction in 5 samples indicated by at least 20-fold higher levels of mRNA of IL1B, IL6, TNFA and CD68 compared to the other 18 samples and concomitant with high expression levels of SLC5A1, SLC5A2, AT1R, CYBA, NCF1, ICAM1, VCAM1, MMP2, MMP9 and TGFB1 in contrast to low levels of NOS3. In addition, increased levels of oxidative stress were observed in the same biopsies, which were diminished by the antioxidant N-acetylcysteine (NAC), NADPH oxidase inhibitor (VAS-2870), TNF-α receptor neutralizing antibody (infliximab), ACE inhibitor (perindoprilat), AT1R antagonist (losartan), dual SGLT1/2 inhibitor (sotagliflozin) and selective SGLT2 inhibitor (empagliflozin) with inhibitory effects reaching up to 80%. Immunofluorescence staining indicated signals for nitro-tyrosine, TNF-alpha, SGLT1 and 2 in several samples.

Conclusion

These findings indicate that both isoforms SGLT1 and SGLT2 are expressed in the transplanted human heart and suggest a pattern of expression associated with pro-inflammatory response. They further indicate a potential protective effect of SGLT2 inhibitors in transplanted hearts through mitigating oxidative stress and hence providing a possible novel therapy for heart transplantation recipients to preserve the heart function.

Funding Acknowledgement

Type of funding sources: Private company. Main funding source(s): Boehringer Ingelheim Pharma GmbH & Co. KG
A Human atlas of smooth muscle cell gene expression; insights from the FANTOM consortium CAGE dataset
Authors: Vaja R , Atanur S , Mitchell J A , Ahmetaj-Shala B
Published: October 2022

Background

Under normal physiological conditions, smooth muscle cells (SMC's) have a role in both vessel contraction and generation of the extra-cellular matrix. It has now become apparent that SMCs throughout the vasculature play a key role in several cardiovascular conditions including hypertension, atherosclerosis, vasculopathy and myocardial infarction. However, how SMC gene expression compares in different vascular and non-vascular anatomical regions remains unknown.

Purpose

To create a human SMC gene expression map using data from the FANTOM consortium Cap-analysis gene expression (CAGE) dataset, a primary database of almost 600 human primary cell samples.

Methods

The gene level expression data was obtained by aggregating expression levels of all the promoters belonging to same gene as defined by the FANTOM CAGE dataset. The anatomical regions (n: 13) included the following vascular sites: aortic, brachiocephalic, carotid, coronary, internal thoracic & pulmonary arteries. Non-vascular regions included colon, oesophagus, prostate, uterine, trachea and bronchial. Weighted correlation network analysis (WGCNA) was used as an unsupervised approach to identify clustering of gene sets.

Results

Hierarchical clustering (Figure 1) demonstrates a distinct clustering of vascular SMC gene sets (aortic, carotid, coronary, internal thoracic, pulmonary & subclavian arteries). The same pattern of vascular SMC clustering was also seen in WGCNA (Figure 2). Gene ontology enrichment of vascular SMCs that clustered in WGCNA identified common gene sets relating to cytokine and chemokine activity. KEGG pathway analysis highlighted several pathways linked to inflammation (TNF, IL17 & NF-κB signalling) as well as pathways linked to atherosclerosis and lipid metabolism.

Conclusion

Vascular SMCs distinctly cluster away from non-vascular SMC types in humans and the separation is primarily driven by gene sets relating to inflammation, lipid metabolism & atherosclerosis. Mapping SMC gene expression at various anatomical regions in both healthy and diseased states may give us better insight into the role of SMCs in disease progression and could identify novel mechanistic targets for future therapy.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): British Heart Foundation
Dysregulation of cardiac endothelial lncRNA H19 in COVID19 patients induces endothelial dysfunction, which impairs cardiomyocyte function
Authors: Juni R P , Phelp P , Bink D , Espinoza-Gonzales P , Van Bergen A , Kuster D , Van Der Velden J , Krijnen P , Niessen H , Boon R
Published: October 2022

Background

COVID19 is accompanied by cardiac complications. Long non-coding RNAs (lncRNAs) have been implicated in the pathogenesis of cardiovascular diseases. However, their contribution to the cardiac manifestation of COVID19 is unknown.

Methods and results

We discovered that endothelial-enriched lncRNA H19 is downregulated in the heart of patients with COVID19 (∼2 fold, p<0.01). H19 was highly expressed in cardiac microvascular endothelial cells (CMEC) as compared to the other endothelial cell types (∼10 fold, p<0.05), suggesting its cardiac enrichment. H19 silencing in CMEC induced endothelial stress phenotype and a reduction in endothelial markers VE-cadherin and eNOS (∼1.5 fold, p<0.01), indicating its importance in endothelial physiology. Using the endothelial-cardiomyocyte co-culture system we previously developed, we showed that H19 silencing in CMEC reduced cardiomyocyte (CM) relaxation and contraction (∼1.5 fold, p<0.01). Interestingly, exposure to plasma from COVID19 patients decreased endothelial H19 level and impaired endothelial enhancement of CM function. Mechanistically, reduced level of H19 increased endothelial IL6 expression (∼1.5 fold, p<0.01). Further, exposure of CMs to IL6 also impaired CM relaxation and contraction, suggesting that endothelial cells devoid of H19 release IL6 which represses CM function. Interestingly, we found increased IL6 levels in the heart of COVID19 patients (∼2 fold, p<0.05). Indeed, the impairment of endothelial enhancement of CM function upon H19 silencing in CMEC was restored in the presence of tocilizumab, an IL6 receptor antagonist (∼1.5 fold, p<0.01). Furthermore, the impairment of the endothelial control on CM function upon exposure to COVID19 plasma was mitigated when the patients were treated with tocilizumab (∼1.5 fold, p<0.01).

Conclusion

COVID19 reduces cardiac endothelial H19 level and induces impairment of endothelial enhancement of CM function via increased release of endothelial-derived IL6, the effect that can be rescued in the presence of IL6 receptor blocker tocilizumab.

Funding Acknowledgement

Type of funding sources: Public grant(s) – EU funding. Main funding source(s): ERC
Plasma eicosanoid profiling in the course of proprotein convertase subtilisin-kexin type 9 inhibition: insights from a metabolomic analysis
Authors: Schrutka L , Hagn G , Galli L , Poeschl A , Seidl V , Ondracek A , Bileck A , Lang I , Hengstenberg C , Krychtiuk K , Speidl W , Gerner C , Distelmaier K
Published: October 2022

Background

Treatment with monoclonal antibodies targeting circulating proprotein convertase subtilisin-kexin type 9 (PCSK9) was found to reduce all-cause mortality in addition to cardiovascular events, suggesting pleiotropic effects. Eicosanoids are bioactive metabolites involved in cardiovascular disease and have not yet been studied in the course of PCSK9 inhibition.

Methods

In this prospective translational single-center study, plasma samples were collected from 64 patients before and after initiation of PCSK9 inhibitor treatment. Metabolomic analyses were performed using liquid chromatography coupled to high-resolution mass spectrometry.

Results

A total of 62 bioactive eicosanoids were detected. Among the metabolites, four were significantly decreased by PCSK9 inhibition after one month and remained stable after 6 months (figure): arachidonic acid (p=0.003), 12,13-DiHOME (p<0.001), 9-HpODE_9.91 (p=0.007) and HpODE_7.71 (p=0.011). Phospholipase A2 levels were reduced by 40% after 1 month (p=0.003) and by additional 50% after 6 months of treatment (p=0.015), but did not correlate with eicosanoids (p=0.057). The change in arachidonic acid levels resulted in a significant increase in the ratio of omega-3 to omega-6 polyunsaturated fatty acids (p=0.002).

Conclusion

PCSK9 inhibition leads to significant changes in the eicosanoid profile already after one month, in particular to a downregulation of arachidonic acid. This discovery complements the presumed pleiotropic effects of PCSK9 inhibition and may provide additional benefit in the treatment of atherosclerotic disease.

Funding Acknowledgement

Type of funding sources: None.
Autoantibodies may predict the neurological severity of patients with Wilson disease
Authors: Wolska M , Eyileten C , Nowak A , Palejko A , Cudna A , Litwin T , Antczak-Kowalska M , Kurkowska-Jastrzebska I , Czlonkowska A
Published: October 2022

Background

Wilson's disease (WD) is a rare, autosomal disorder of copper metabolism, in which antibodies mediate and modulate the clinical manifestation of the CNS disease. The aim of this study was to investigate the presence and impact of autoantibodies in the clinical course of WD.

Method

Study involved 88 WD diagnosed and already treated patients with 7-years follow-up. Indirect immunofluorescence assays and Western Blot analysis was used.

Results

Patients who presented neurological manifestations had significantly more frequently ANA, ANCA and ANNA presence than in healthy individuals (p=0.029, p=0.009, p=0.025, respectively). In patients treated with d-penicillamine positive ANA antibodies were observed more frequently (p=0.005), whereas in zinc sulphate treated patients PP, ANCA and ANNA were significantly more frequently positive (p=0.038, p=0.024, p=0.002, respectively). Within the group of patients who died, positive ANA antibodies were significantly more frequent in comparison to patients who survived (p=0.020). Patients presenting positive cANCA had significantly higher UWDRS II, III and total scores compared to patients without ANCA antibody (p=0.033, p=0.022, p=0.022, respectively).

Conclusion

Drug-induced ANCAs may have a predictive value, as it is positively correlated with UWDRS II, III and total score in patients with WD. Only patients with neurological manifestations had significantly higher ANA, ANCA and ANNA presence. Increased positivity of those antibodies may predict the neurological manifestation of the severe cases and may be helpful for the neurological assessment of WD.

Funding Acknowledgement

Type of funding sources: Public Institution(s). Main funding source(s): Medical University of Warsaw
Positron emission tomography/computed tomography perfusion imaging with myocardial blood flow has diagnostic and prognostic value for cardiac allograft vasculopathy
Authors: Abadie B , Chan N , Sharalaya Z , Bhat P , Harb S , Jacob M , Tang W H , Cremer P , Jaber W
Published: October 2022

Background

Cardiac allograft vasculopathy (CAV) is a leading cause of morbidity and mortality in patients with orthotopic heart transplantation (OHT). Invasive coronary angiography is the traditional method of screening for and diagnosing CAV. Alternative non-invasive modalities have been sought to screen for CAV. A small, single-center study utilizing Positron Emission Tomography/Computed Tomography Perfusion Imaging (PET/CT) with stress myocardial blood flow (MBF) demonstrated good diagnostic and prognostic value for CAV.

Purpose

The purpose of this study was to validate the proposed algorithm for diagnosing and prognosticating CAV by PET/CT with stress MBF in a large and contemporary series.

Methods

Patients with a history of OHT with no prior revascularization in the transplanted heart who underwent PET/CT with MBF were included in the prognostic portion of the study. For the diagnostic value of PET/CT with MBF, only patients who had a PET/CT within 12 months of coronary angiography were included. The diagnostic accuracy of PET/CT was compared to the most recent coronary angiogram. A composite outcome of death, heart failure hospitalization, acute coronary syndrome, revascularization, and re-transplantation was used to validate the prognostic ability of PET/CT with MBF.

Results

450 PET/CT scans with MBF were performed and included in the prognostic portion of the study. 78 patients had PET/CT within 12 months of coronary angiography and were included in the diagnostic portion. Normal perfusion with normal myocardial blood flow had a 100% negative predictive value for moderate-severe CAV by angiography. PET/CT CAV 2/3 had a positive predictive value of 69% for moderate-severe CAV. Over 24 months, there were 20 events in the 39 patients with a PET CAV grade of 2/3 versus 21 events in the 411 patients with a PET CAV grade of 0/1 with a hazard ratio 13.3 (p<0.001).

Conclusions

The current proposed algorithm for diagnosing CAV by PET/CT with stress MBF has excellent negative predictive value along with good positive predictive value for detecting moderate-severe CAV by coronary angiography. A PET/CT with stress MBF with CAV classification of 2/3 is associated with a poor prognosis.

Funding Acknowledgement

Type of funding sources: None.