bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of cardiovascular events on primary and hospital care costs: findings from UK Biobank study
Authors: Zhou J , Wu R , Williams C , Emberson J , Reith C , Keech A , Robson J , Wilkinson K , Armitage J , Collins R , Gray A , Simes J , Baigent C , Mihaylova B
Published: October 2022

Background

Need for primary and secondary healthcare increases following cardiovascular disease (CVD) events but there is no data on comparative increases in costs.

Purpose

To estimate annual primary care and hospital inpatient costs associated with key CVD and other adverse events using the UK Biobank (UKB) individual participant data.

Methods

UKB participants with linked primary care data (192,983 participants) or hospital inpatient episodes data (all 501,807 participants) contributed data to this study. The three categories of primary care services (patient consultations, diagnostic and monitoring tests, prescription medications), and hospital episodes were costed (2020 UK£) using the NHS England reference costs. Annual primary care costs and, separately, annual hospital inpatient costs were modelled as functions of participant characteristics at entry (socio-demographic, clinical, prior diseases) and time-updated first occurrences of myocardial infarction, stroke, coronary revascularization, incident cancer, incident diabetes, vascular death and non-vascular death during follow-up (p-value <0.01 in stepwise covariate selection). One-part generalized linear regression model (GLM) with Poisson distribution and identity link function was used for primary care costs, and two-part model was used for inpatient costs (part 1: logistic regression models probability of incurring costs; part 2: GLM with Poisson distribution and identity link function models costs conditional on incurring any). Separate models were fitted among participants with and without previous CVD at entry into UKB.

Results

Most adverse events were associated with excess primary care and hospital inpatient costs. Compared to people without previous CVD, people with previous CVD had on average larger excess primary care and hospital inpatient costs in years with myocardial infarction, stroke and vascular death; but similar excess costs in years with other events. Among both people without and with previous CVD, the excess annual primary care costs were less than 7% of the excess annual hospital inpatient costs for vascular events (Table). However, following diabetes diagnosis the excess annual primary care costs were higher than the excess annual hospital inpatient costs (Table).

Conclusions

These excess primary and hospital care costs associated with CVD events could inform assessments of interventions and policies to reduce CVD risks in UK.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): UK National Institute for Health Research (NIHR) Health Technology Assessment (HTA) Programme, UK Medical Research Council (MRC), British Heart Foundation
Dutch Nationwide Implementation Program for familial hypercholesterolemia case finding in children: a cost-effectiveness and return on investment analysis
Authors: Ademi Z , Norman R , Sijbrands E , Ference B A , Watts G F , Hutten B , Wiegman A
Published: October 2022

Background

The Netherlands is one of the few countries in the world that actively screen for familial hypercholesterolemia (FH).

Objectives

We investigated the return on investment (ROI) and cost-effectiveness of the Dutch nationwide implementation program for FH case-finding and preventive treatment in children compared to later detection and treatment from both a societal and healthcare perspective in the Netherlands.

Methods

Cascade case finding and early preventive treatment were modelled to simulate the progression of disease and costs of ten-year-olds suspected of having heterozygous FH (HeFH) over a lifetime. The model consisted of three health states: alive without coronary heart disease (CHD), alive with CHD, and dead. The decision was to compare the nationwide implementation program for FH cascade case-finding in children and immediate treatment with statins compared to “usual care” later detection and treatment. The prevalence of HeFH in this target population was 51.2% as per the Dutch Health Program for FH case-finding, and the sensitivity and specificity of testing was 100%. Contemporary Mendelian Randomisation Analysis data was used to quantify the risk of a first coronary heart disease event as a function of age and total lifetime exposure to low-density-lipoprotein cholesterol. Costs and outcome were sourced from the Dutch data and other published sources. ROI for the Dutch nationwide implementation program for FH case-finding and cost-effectiveness analyses using incremental cost-effectiveness ratios (ICERs) (cost per quality-adjusted life years (QALYs) gained), compared with current usual care were the main outcomes of interest. All future benefits and costs were discounted annually by 1.5% and 4% respectively.

Results

Over the lifetime horizon, the ROI for the Dutch nationwide implementation program for FH case-finding was €16.52 (i.e. every euro spent on the program resulted in a return of €16.52). From the societal perspective, the Dutch nationwide implementation program for FH case-finding and early treatment with statins was the dominant strategy (health improvement and cost saving) compared to usual care. From a healthcare perspective, the Dutch nationwide implementation program for FH case-finding and early treatment with statins would gain 2.64 QALYs per person, at an additional cost of €7,178 (both discounted). These equated to an ICER of €2,668 per QALY gained.

Conclusions

From the societal perspective, the the Dutch nationwide implementation program for FH case-finding represented excellent value for investment for Netherlands, being not only health saving but also cost saving form a societal perspective and cost effective from a healthcare perspective.

Funding Acknowledgement

Type of funding sources: None.
Long-term effects of coronavirus disease 2019 on the cardiovascular system: the CV COVID-19 registry
Authors: Arevalos Rivas V , Ortega-Paz L , Fernandez-Rodriguez D , Jimenez-Diaz V A , Baneras Rius J , Campo G , Diaz J F , Scardino C , Rodriguez-Santamarta M , Gonzalo N , Perginotti A , Alfonso F , Amat-Santos I , Sabate M , Brugaletta S
Published: October 2022

Background

Patients with COVID-19 have an increased risk of cardiovascular adverse events during the acute phase. However, the long-term cardiovascular outcomes are unknown.

Objective

We aimed to determine the long-term effects of COVID-19 in the cardiovascular system.

Methods

This is a multicenter, observational, retrospective registry conducted at 17 centers in Spain and Italy. Consecutive patients older than 18 years who underwent a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for SARS-CoV2 in the participating institutions were included. Patients were classified into two groups, according to the results of the RT-PCR: COVID-19 positive or negative. The primary outcome was cardiovascular (CV) death at 1-year. The secondary outcomes included acute myocardial infarction, stroke, heart failure hospitalization, pulmonary embolism, and serious cardiac arrhythmias at 1-year. Outcomes were compared between the two groups. An independent clinical event committee adjudicated events.

Results

A total of 4427 patients were included, 3578 (80.8%) patients with COVID-19 and 849 (19.2%) without COVID-19. COVID-19 patients were older, had a higher rate of classical cardiovascular risk factors, except for active smoking, and had fewer comorbidities. At a median time of 13.5 (IQR 11.8–15.8) months, after an adjustment by baseline characteristics, there was no difference in CV death (1.4% vs. 1.1%; HR 1.03 [0.49–2.18]; p=0.941) between patients with COVID-19 and without. However, COVID-19 patients experienced higher rate of venous thromboembolism (VTE) (3.9% vs. 0.6%, HR 6.11 [2.46–15.16]; p=0.001), major bleeding (2.9% vs. 0.5%, HR 5.38 [1.95–14.84]; p=0.001), and serious cardiac arrhythmias (2.6% vs. 0.9%, HR 2.25 [1.07–4.73]; p=0.033). During follow-up, between discharge and end of follow-up, COVID-19 patients did not experience a higher risk of adverse cardiovascular outcomes (composite of CV death, any MI, ischemic stroke, systemic arterial thrombosis, VTE, heart failure hospitalization, or any serious arrhythmia) compared to patients without (HR 0.80; [0.53–1.21]; p=0.298).

Conclusions

At 1-year follow-up, COVID-19 was not associated with an increased risk of cardiovascular death but with a higher risk of VTE events, major bleeding, and serious cardiac arrhythmias. COVID-19 was not associated with a higher risk of adverse cardiovascular events during follow-up.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Carlos III Institute, Madrid, Spain
Causes of admission and mortality among patients admitted in 37 cardiology departments in 17 Sub-Saharan African countries: the FEBRUARY observatory (2016–2021) from African Research Network
Authors: Ali Toure I , Traore A K , Cavagna P , Damorou J M , Diao M , Dzudie A , N'Guetta R , Kouam Kouam C , Hermann Y , Limbole E , Gaye B , Toure C , Yameogo V , Jouven X , Antignac M
Published: October 2022

Background

Cardiovascular disease (CVD) is a major cause of death worldwide. At least two-third of CVD death occurs in low- (LIC) and middle-income (MIC) countries. An estimated 1 million deaths were attributable to CVD in sub-Saharan Africa (SSA) alone. Scarce data are available about the admission for CVD and outcome in cardiology department in SSA.

Purpose

To describe admission for CVD and outcome of hospitalization in 17 in SSA countries.

Methods

We conducted a transversal and longitudinal study in CV department of 37 hospitals from 23 cities in 17 SSA countries (10 low income: Niger, Guinea, Benin, Mali, Democratic Republic of the Congo, Tchad, Burkina Faso, Togo, Burundi, Ethiopia and 7 middle income: Cote d'Ivoire, Senegal, Cameroon, Congo, Soudan, Mauritania, Gabon). The February study was designed by a multidisciplinary collaborative team of epidemiologists, pharmacists and cardiologists from Africa and France. This ongoing observatory included all inpatients in February from each year since 2016.Data including socio-demographic and clinical characteristics, causes of admission, clinical, biological, complementary examinations, treatments, length of stay and discharge diagnosis were collected by the investigating physicians. All analyses were performed through scripts developed in the R software (4.0.3 (2020-10-10))

Results

Overall, 4360 patients were included in the February study. Men represented 56.4%. Mean of age was 56.7±16.8 years. The main cause of admission was heart failure (41.5%) followed by acute coronary syndrome (11.9%) and stroke (11%). Cause of admission varied significantly across countries (p<0.01) and over the years (p<0.01). Proportions of admission for heart failure varied from 20% in Ethiopia to 62% in Guinea. Mean of length of stay was 9.95±22.4 days. All causes of admission confounded, in-hospital mortality rate was 11% (N=423). Mortality rate differed significantly according to causes of admission (p<0.01).Mortality rate among patients admitted for stroke, heart failure and acute coronary syndrome was respectively 18%, 12.3% and 9.5%. Overall, 31 (0.7%) were admitted for endocarditis and mortality rate among them was 20.8% (figure). Overall, mortality rate did not vary according to gender or over the years but was significantly different according to patient wealth index (p<0.05), countries (p<0.01) and level income countries (p<0.01). Mortality rate was significantly higher in LIC.

Conclusion

Heart failure, acute coronary syndrome and stroke represent the two-third of causes of admission. Overall, mortality rate in cardiology departments in SSA reached 11%.

Funding Acknowledgement

Type of funding sources: None.
Accuracy of event rate and effect size estimation in major cardiovascular trials
Authors: Olivier C B , Struss L , Suennen N , Kaier K , Heger L A , Meerpohl J J , Mahaffey K W
Published: October 2022

Background

During the design of a randomized controlled trial (RCT), the estimation of the expected event rate (ER) and effect size (ES) of an intervention is an essential step to calculate the sample size, which determines the scope of the trial. Overestimation of ER and ES might lead to an underpowered trial design.

Objective

To evaluate the accuracy of the estimation of ER and ES in contemporary cardiovascular RCTs.

Methods

We searched “Medline” for cardiovascular RCTs published in the New England Journal of Medicine, The Journal of the American Medical Association, or The Lancet between 2010 and 2019. Only multicenter RCTs with a dichotomous primary endpoint and with not more than 2 study arms were included. Data were extracted from the original publication or the associated study protocol. For trials that tested superiority of an intervention, the effect size measurements hazard ratio, odds ratio, or risk ratio were summarized as the effect size ratio (ESR). To determine the accuracy of estimation, we compared the observed to the hypothesized ER and ESR, respectively. The association of the accuracy of estimation with trial characteristics was determined by linear regression.

Results

Of 875 identified publications, 374 underwent full review. After exclusion of trials with insufficiently reported data for this analysis, data from 344 trials were analyzed. 145 trials (42.2%) were industry sponsored, 122 (35.5%) were investigator-initiated with industry-co funding, and 77 (22.1%) with government or public funding. The majority (186; 54.1%) tested a drug as the intervention. 263 (76.5%) were designed to test for superiority of the intervention. The primary endpoint consisted of at least 2 components in 247 (71.8%) trials. The median estimated sample size was 2386 (interquartile range [IQR] 800, 6000).

The median observed event rate was 9.0% (IQR 4.3, 21.4%) and was significantly lower compared with the estimated ER (11.0% [IQR 6.0, 25.0%]; median deviation from estimation −6.9% [95% confidence interval, CI −10.0, −3.7%]; p<0.001). 196 trials (61%) overestimated the event rate (Figure 1A). The overestimation was consistent across several subgroups (Figure 2A). The median observed ESR was 0.910 (IQR 0.740, 0.990) and was significantly higher compared with the estimated ESR (0.724 [IQR 0.600, 0.795]; median deviation from estimated effect size 26.0% [95% CI, 19.0, 33.0%]). 216 (82.1%) trials overestimated the effect size (Figure 1B). This overestimation was consistent across all analyzed subgroups (Figure 2B) and was less pronounced in trials with use of a composite primary outcome.

Conclusion

6 of 10 contemporary cardiovascular RCTs overestimated the event rate for the primary endpoint and 8 of 10 trials overestimated the effect sizes of an intervention. Overestimation may lead to inability to answer the trial hypothesis.

Funding Acknowledgement

Type of funding sources: None.
Academic spam mails from “Predatory Journals” in cardiology: a cross-sectional study over 3 months
Authors: Ohlow M
Published: October 2022

Background

Numerous online journals are referred to as predatory journals because their efforts are focused primarily on collecting publication fees rather than on content, peer review, or manuscript presentation. Many of these journals send Academic Spam Mails (ASM), requesting manuscript submissions.

Purpose/Methods

Over a 3-month period, all ASMs addressed to the author were collected and analyzed. A fake publication was created with a commonly used structure (Introduction, Methods, Results and Discussion). The article just repeated the phrase “Please get me off your mailing list” over and over again. This publication was submitted in response to each request to test the control mechanisms of the respective journals.

Results

During the study period, excluding duplicate requests, 125 ASMs (average 2.1 per business day) were received from a wide variety of journals spanning 38 different subject areas, of which 29/38 (76.4%) were from the medical field. 16/125 (16%) were related to a previously published article but not among the author's most important publications (median h-index rank 22; range 10–138). The majority (73.6%) of journals were listed in Beall's predatory journal list, only an absolute minority (0.8%) were listed in the Directory of Open Access Journals, and 96% were not listed in PubMed. Original papers (94.4%), reviews (88.0%), and case reports (87.2%) were most frequently requested; the publication charge (APC) was a median of $1,199 (range $50–3,586); in about 22% of cases, it was not apparent whether APCs were charged and, if so, how much. For the 125 nonsense publications sent out, there was a response in 16/125 (13%) (6 acknowledgements of receipt, 3 apologies, 2 the journal comments that the publication consisted of only one sentence, and 5 other responses). Interestingly, in one case editorial board members had also created a nonsense publication titled: “Ok we not send mail to you” and sent it to the author. 21.6% of the journals had sent ASM to the author again during the next months.

Conclusions

Academic spam mails from “predatory” online journals are a common problem. They are recognizable by their appearance in warning lists or non-appearance in directories of reputable open access journals. A placement of a nonsense publication in such a “predatory journal” was not successful.

Funding Acknowledgement

Type of funding sources: None.
ZNF259 rs964184 variant is associated with dyslipidemia and coronary artery disease in the young population
Authors: Santos M , Mendonca M I , Sa D , Temtem M , Sousa A C , Henriques E , Rodrigues M , Freitas S , Borges S , Guerra G , Freitas A I , Ornelas I , Drumond A , Palma Dos Reis R
Published: October 2022

Introduction

Coronary artery disease (CAD) is a dynamic inflammatory disease caused by atherosclerosis. GWAS showed that ZNF259 rs964184 encoding zinc finger protein (ZPR1) was associated with dyslipidemia and CAD. Recent research found that ZPR1 transcription is up-regulated in the brain of mice fed a high-fat diet, influencing the cell cycle, apoptosis, and RNA metabolism in neurons. This process at the heart vessels may increase oxidative stress and CAD.

Purpose

Study the association between the ZNF259 rs964184 C>G polymorphism with dyslipidemia and CAD susceptibility in a Portuguese population.

Methods

A case-control study was performed with 3,160 individuals, namely 1,723 CAD patients (mean age 53.3±7.9; 78.7% male) and 1,437 controls (mean age 52.8±7.8; 76.3% male). Participants were stratified into two age groups (<45 and >55 years). ZNF259 rs964184 C>G was genotyped and analysed using the dominant model (CG+GG vs CC). Multivariate logistic regression was performed in both age groups to investigate whether rs964184 polymorphism was associated with dyslipidemia and CAD susceptibility.

Results

The dominant model of ZNF259 was associated with dyslipidemia (OR=1.85; 95% CI: 1.22–2.79; p=0.003) and CAD (OR=1.46; 95% CI: 1.02–2.09; p=0.036) in the younger population under 45 years. In the >55 years group, this model was associated with dyslipidemia (OR 1.46; 95% CI: 1.06–2.01; p=0.020) but not with CAD. After multivariate logistic regression, the CG+GG remained an independent risk factor for CAD susceptibility only in the population <45 years (OR=1.60; 95% CI: 1.03–2.50; p=0.037).

Conclusion

ZNF259 rs964184 is a risk factor for dyslipidemia in the whole population. Dyslipidemia may up-regulate ZPR1 transcription, enhancing the vulnerability of coronary endothelial cells to both oxidative stress and inflammatory response, increasing CAD susceptibility. This mechanism seems more relevant at the cellular level in young patients representing a possible prophylactic and therapeutic target, especially in this age group.

Funding Acknowledgement

Type of funding sources: Public hospital(s). Main funding source(s): SESARAM EPERAM
Syndromic cardiac disorder is associated with a non-coding deletion that induces a 3D chromatin remodeling and PITX2 expression dysregulation
Authors: Baudic M , Murata H , Bosada F M , Souto Melo U , Aizawa T , Guedon A , Lindenbaum P , Gourraud J B , Makita N , Mundlos S , Christoffels V M , Makiyama T , Probst V , Schott J J , Barc J
Published: October 2022

In a first family (family#1), we identified 53 members of whom 17 present a syndromic cardiac disorder characterized by electrical disorders (sinus node dysfunction, atrial fibrillation...) and developmental defects (atrial septal defect, valvopathy, left ventricle non-compaction...) following an autosomal dominant model. Among the affected family members, 6 are implanted with a pacemaker and one experienced a sudden death at 43yo. Despite a strong linkage pointing to the 4q25 region, exome sequencing failed to identify causal variant. Interestingly, 6 additional non-related families presenting the same phenotype have been also identified.

Our aims are to identity the causal mutation and the molecular mechanism underlying this complex cardiac syndrome.

Genetic study has been performed using whole genome sequencing (WGS). Based on transgenic mouse strains, we assessed the impact of Family#1 mutation on the phenotype and on gene expression. Then, we generated human cardiomyocytes derived iPS cells (CM-iPS) isogenic models to evaluate the epigenome (CUT&RUN and ATAC-seq), transcriptome (RNA-seq) and topological associated domain (TAD) remodelling (Hi-C).

By WGS we uncovered a deletion of 15kb in a gene desert area on 4q25, segregating in all affected relatives of Family#1. The 6 other families present overlapping deletions. Mouse model recapitulates the cardiac phenotype and exhibit a dysregulation of Pitx2 expression in cardiac specific compartments. Based on human CM-iPS models, epigenetic data highlight among the 15kb deletion a unique open region containing a CTCF binding site, crucial for delimiting TAD boundaries. Hi-C assay reveals the fusion of 2 TADs and highlights new interactions between PITX2 and atrial specific regulatory elements.

We identified a deletion located within a gene desert area associated with a complex cardiac disorder. The CTCF binding site contained in the deletion seems key in the TAD border. The TAD remodelling leads to new (regulatory) interactions and expression dysregulation of PITX2. We describe a new molecular mechanism implying a yet unidentified non-coding regulatory element of PITX2 and responsible for a complex electrical and developmental cardiac syndrome.

Funding Acknowledgement

Type of funding sources: Public Institution(s). Main funding source(s): Nantes UniversitéFrench national reserch agency (ANR)
PDE1 inhibition and adenosine A2A receptor activation synergistically potentiates spontaneous calcium waves and transients in mouse ventricular myocytes
Authors: Casabella-Ramon S , Jimenez-Sabado V , Hove-Madsen L
Published: October 2022

Background

The phosphodiesterase-1C (PDE1C) and the adenosine A2A receptor (A2AR) are located in a macromolecular cluster and expected to modulate cAMP-dependent regulation of the intracellular calcium homeostasis in cardiac myocytes. In this contect, atrial fibrillation has been associated with a high incidence of spontanoeus arrhythmogenic calcium waves caused by excessive A2AR activation. On the other hand, inhibition of PDE1C and A2AR activation has also been reported to produce a synergistic and protective effect on cardiomyocyte survival, but little is known about the corresponding effects on the intracellular calcium homeostasis.

Purpose

This study, therfore, aimed to investigate how PDE1 inhibition and/or A2AR activation modulate sarcoplasmic reticulum (SR) calcium homeostasis in mouse ventricular myocytes.

Methods

Mouse ventricular myocytes (47 cells from 20 mice) were loaded with the calcium indicator Rhod-2 and spontaneous calcium release events at rest or caffeine induced calcium transients were visualized using resonance-scanning confocal microscopy at a frame rate of 150 Hz. The SR calcium content was estimated as the ratio of the peak fluorescence and the fluoresence the end of a 10 mM caffeine pulse (20s). Calcium leak was estimated as ratio of the flourescence emmision at baseline before and at the end of the caffeine pulse.

Results

The incidence of spontaneous calcium waves or transients increased from 0.7±0.3 events/min in control to 1.1±0.5 events/min and 3.4±1.0 events/min (p<0.01) after inhibition of PDE1 with the selective antagonist ITI-214 and the A2AR agonist CGS-21680 respectively. Concomitant PDE1 inhibition and A2AR activation synergistically increased the event frequency to 8.4±2.3 events/min (p<0.05). There were no significant differences in the corresponding SR calcium contents (4.4±0.3, 4.5±0.4, 4.7±0.4 and 4.1±0.2 for control, ITI-214, CGS-21680 and ITI-214+CGS-21680 respectively). However, there was a parallel increase in calcium leak from 8.7±4.2% in control to 21.8±9.8% with ITI-214, 17.6±7.2% with CGS-21680 and 37.5±11.1% when myocytes were exposed to both compounds (p<0.05), suggesting that the higher incidence of spontaneous calcium waves and transients observed upon PDE1 inhibition and/or A2AR activation is likely caused by a lowering of the threshold for spontaneous SR calcium release rather than by SR calcium overloading.

Conclusion

PDE1 inhibition exacerbates the incidence of spontaneous arrhythmogenic SR calcium release events in myocytes with high levels of A2AR activation, suggesting that PDE1 inhibition under these conditions increases the risk of arrhythmia.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Spanish Ministry of Science and Innovation
Cardiomyocytes derived small extracellular vesicles plays an important role in heart development
Authors: Balbi C , Ceresa D , Schiano C , Malatesta P , Barile L , Vassalli G
Published: October 2022

Introduction

Neonatal rats have the capacity to regenerate their hearts in response to injury, but this potential is lost after the first week of life. Cardiac maturation lays the foundation for postnatal heart development and disease, yet little is known about the contributions of the microenvironment to cardiomyocyte maturation.

Extracellular vesicles (EV) are bilayer-membrane nanoparticles released by all cell types, carrying proteins, lipids, and nucleic acids, which reflect the activation state of parental cells. Secreted small extracellular vesicles (sEV), prominently figure among extracellular signals that regulate cell function.

Purpose

We aim to determine whether cardiomyocyte (CM) derived sEV carried miRNA that has a role in heart development with specific regards to cardiomyocyte maturation.

Methods

sEV were isolated from rat cardiomyocyte at day 0 and 7 after birth by serial ultracentrifugation. sEV were characterized by NTA and analyzed by Western blot for the presence of classical EV markers (TSG101, Syntenin-1). The role of sEV in cardiomyocyte proliferation was assessed by analysis of EdU incorporation on neoantal rat CM treated with sEV_p0 or sEV_p7. miRNA content on sEV was assessed using a rat-miRNome MicroRNA Profiling Kit and the identified miRNA's targets confirmed by RealTime-PCR and Western Blot.

Results

NTA and Western Blot analysis confirmed the presence of sEV in both the extracellular vesicles preparation. sEV_p0 showed to be able to sligtly increase EdU incorporation in treated cardiomyocyte (1.15-Fold) while sEV_p7 significantly inihibit CM proliferation (0.78-Fold) toghether with a change in cardiomyocyte citoscheletal architecture. Data from miRNome analysis showed in sEV_p7 a significan increase in miRNA with cyclines as tagets. Downregulation of Cdk1; Cdk4; Cdk2; CcnB1 and CcnD1 was confirmed on CM trated with sEV_p7 compared to Ctrl. Downregulation of CCND1 was aslo confrmed at the protein level by Western Blot analysis.

Conclusions

These preliminary resultes showed an important role in heart develompent of cardiomyocyte derived-sEV. A deeper investigation of the pathaway activated by sEV may have a potential interest for the identification of possible regulators for stimulating heart regeneration.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): Fondazione Leonardo, Lugano, Switzerland
Programmed cell death protein 1 (PD1) deficiency induces cardiac inflammation during baseline conditions and ischemia/reperfusion injury
Authors: Michel L , Korste S , Spomer A , Stock P , Odersky A , Hendgen-Cotta U B , Rassaf T , Totzeck M
Published: October 2022

Background

The Programmed cell death protein 1 (PD1) immune checkpoint is densely expressed on cardiac endothelial cells. The increasing clinical application of immune checkpoint inhibitor therapy targeting PD1 for the treatment of advanced malignancies has revealed profound cardiovascular side effects that have recently paralleled in a preclinical model. However, the effects of PD1 deficiency during baseline conditions and myocardial injury are so far unknown.

Purpose

This study aims to assess the impact of PD1 deficiency on myocardia immunity, and to evaluate the relevance of PD1 signalling in cardiac disease.

Methods

C57BL/6J wild-type mice and C57BL/6J Pdcd1−/− mice were purchased and bred at the animal facility. In-vivo ischaemia/reperfusion (I/R) injury was applied to assess the response in cardiac injury. In brief, mice were anesthetised followed by lateral thoracotomy and ligation of the left coronary artery for 45 min. For flow cytometry, hearts were removed and subjected to an enzymatic digestion. Single cell solutions were stained with different antibody panels to assess cardiac immune cells and the expression of programmed cell death protein 1 ligand 1 (PDL1). Western blot was conducted after homogenization of snap-frozen heart tissue using specific primary and corresponding secondary antibodies. Immunofluorescence and conventional haematoxylin and eosin stain from 4 μm sections were used to visualize the distribution of PDL1 in cardiac tissue.

Results

While no overt cardiac phenotype was observed in Pdcd1−/− mice, a profound upregulation of pro-inflammatory cytokines was determined during baseline conditions, including interleukin (IL) 1α IL4, and extracellular-signal regulated kinase (ERK) 1/2. NADPH oxidase 1 (NOX1) which is involved in production of reactive oxygen species and endothelial injury response was downregulated in Pdcd1−/− mice. Following I/R injury, a significant decline in endothelial PDL1 expression was observed, which was attributed to changes in the area at risk, as shown by immunofluorescence staining. The infarct size following in-vivo I/R injury was not altered upon PD1-deficiency as determined by TTC staining. However, flow cytometry determined increased cell numbers of distinct leukocyte subsets during reperfusion-related inflammation.

Conclusion

Deficiency of PD1/PDL1 signalling shows distinct inflammatory changes in cardiac tissue at baseline and I/R injury. The results indicate that relevant PD1-related detrimental effects are not limited to complications from cancer therapy but can be expected in various forms of cardiovascular disease, hence requiring further investigations.

Funding Acknowledgement

Type of funding sources: Public hospital(s). Main funding source(s): Department of Cardiology and Vascular Medicine, West German Heart and Vascular Center, University Hospital Essen, Germany
The peri-infarct gray zone of myocardial fibrosis is a better predictor of ventricular arrhythmias than dense core fibrosis in patients with previous myocardial infarction
Authors: Lopes P , Cunha G , Freitas P , Rocha B , Abecasis J , Carmo J , Guerreiro S , Galvao Santos P , Moscoso Costa F , Carmo P , Cavaco D , Morgado F , Mendes M , Adragao P , Ferreira A
Published: October 2022

Background

Current sudden cardiac death (SCD) risk stratification relies heavily on left ventricular ejection fraction (LVEF), but markers to refine risk assessment are needed. Dense core fibrosis (DCF) and peri-infarct “gray zone” of myocardial fibrosis (GZF) on late gadolinium enhancement (LGE) cardiac magnetic resonance (CMR) have been proposed as potential arrhythmogenic substrates. The aim of our study was to determine whether DCF and GZF could predict the occurrence of ventricular arrhythmias in patients with previous myocardial infarction.

Methods

We performed a single centre retrospective study enrolling consecutive patients with previous myocardial infarction undergoing CMR before implantable cardioverter-defibrillator (ICD) implantation. Areas of LGE were subdivided into “core” DCF and “peri-infarct” GZF zones based on signal intensity (>5 SD, and 2–5 SD above the mean of reference myocardium, respectively).

The primary endpoint was a composite of sudden arrhythmic death, appropriate ICD shock, ventricular fibrillation (VF), or sustained ventricular tachycardia (VT) as detected by the device.

Results

A total of 88 patients (median age 61 years [IQR 54–73], 84% male, median LVEF 30% [IQR 23–36%], 14% secondary prevention) were included. During a median follow-up of 23 months [IQR 9–38], 13 patients reached the primary endpoint (10 appropriate ICD shock, 2 sustained VT or VF, and 1 sudden arrhythmic death). Patients who attained the primary endpoint had similar DCF (30.4±14.7 g vs. 28.0±15.3 g; P=0.601) but a greater amount of GZF (18.1±9.6 g vs. 11.9±6.7 g; P=0.005). On univariate analysis, GZF was associated with the composite endpoint (HR: 1.09 per gram; 95% CI: 1.02–1.15; P=0.006), whereas DCF was not (HR: 1.01 per gram; 95% CI: 0.98–1.05; P=0.571). After adjustment for LVEF, GZF remained independently associated with the primary endpoint (adjusted HR: 1.06 per gram; 95% CI: 1.01–1.12; P=0.035). Decision tree analysis identified 11.9g of GZF as the best cut-off to predict life-threatening arrhythmic events. The primary endpoint occurred in 11 out of the 35 patients (31.4%) with GZF ≥11.9 g, but in only 2 of the 53 patients (3.8%) with GZF <11.9 g – Figure 1.

Conclusions

The extent of peri-infarct GZF seems to be a better predictor of ventricular arrhythmias than DCF. This parameter may be useful to identify a subgroup of patients with previous myocardial infarction at increased risk of life-threatening arrhythmic events.

Funding Acknowledgement

Type of funding sources: None.
The long non-coding RNA Heat4 is dynamically regulated during cardiogenic shock
Authors: Winkler M , Kneuer J M , Meinecke T , Moebius-Winkler M N , Weiss R , Haas J , Garfias-Veitl T , Von Haehling S , Keller T , Thiele H , Lurz P , Speer T , Laufs U , Boeckel J N
Published: October 2022

Background and purpose

Cardiogenic shock (CS) remains the leading cause of death in acute myocardial infarction (AMI), with high mortality rates of 40–50%. The long non-coding RNA (lncRNA) Heat4 is associated with the inflammatory response of non-classical monocytes. Previous experimental work shows that this mechanism may be important in heart failure (HF) and during regeneration after vascular injury. Here, we investigate the association of Heat4 with survival in patients with chronic HF and assessed its regulation in AMI and CS.

Methods and results

Heat4 was elevated in the blood of HF patients compared to age-matched non-failing controls (+5.2-fold; HF: N=63; Controls: N=38; p<0.05). Heat4 showed a positive correlation with systemic inflammation (hsCRP; r=0.41; p<0.05) and was negatively associated with LVEF (r=−0.45; p<0.001). Heat4 blood levels showed good discriminatory power for prevalence of HF (AUC = 0.734; p<0.05) and mortality prediction after 4-year follow-up (AUC = 0.789; HF: Death N=32; Controls: Death N=0; p<0.05). Furthermore, Heat4 was elevated in the blood of patients with AMI compared to controls (+1.85-fold; AMI: N=42; Controls: N=23; p<0.05). Heat4 showed a very strong induction in patients suffering from CS (+284.5-fold; CS: N=4; Controls: N=5; p<0.05). In agreement with an anti-inflammatory signaling, Heat4 showed a dynamic regulation in patients with CS with a 284.5-fold increase during acute shock and a decrease 24 hours after revascularization (−82.3% compared to day of revascularization). This regulation was validated in an independent second cohort.

Conclusion

The lncRNA Heat4 is upregulated in the blood of patients with chronic heart failure, acute myocardial infarction and cardiogenic shock. In CS, Heat4 is dynamically regulated. These data set the stage to further assess Heat4 blood levels as a strategy for risk stratification and potential treatment target in HF.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Projektfoerderung im Bereich der Herzmedizin, Leipzig
Multidimensional immunoimaging to characterize the inflammatory reaction after permanent and temporary coronary artery occlusion in mice
Authors: Maier A , Toner Y C , Munitz J , Calcagno C , Fayad Z A , Mulder W J M , Van Leent M M T
Published: October 2022

Background

The immune response following acute myocardial infarction encompasses a delicate balance between inflammatory and reparative programs. Our knowledge of these complex mechanisms is mainly derived from studies using a murine model of permanent coronary artery occlusion. In this study we developed, validated and implemented multiparametric imaging methods to investigate cardiac function and the systemic immune response in transient or permanent coronary artery occlusion mouse models.

Methods

The myocardial infarction models encompassed either transient (40 min) or permanent LAD occlusion in C57BL/6 mice, and non-infarcted mice were used as controls. Two or seven days later, the animals subjected to systemic immunoimaging of the bone marrow, spleen and myocardium with late gadolinium enhancement cardiac MRI (LGE cMRI), 18F-fluorodeoxyglucose (18F-FDG) PET, 18F-Fluorothymidine (18F-FLT) PET, 64Cu-CCR2 PET targeting inflammatory monocytes, 89Zr-CD11b nanobody PET and 19F-HDL-PERFECTA MRI, both targeting myeloid cells. In addition, the same myocardial infarction models were applied to atherosclerosis-prone Apoe−/− and systemic inflammation and plaque progression were assessed by flow cytometry and immunohistochemistry four weeks after infarction.

Results

Through LGE cMRI and 18F-FDG PET, we observed that temporary coronary occlusion resulted in a smaller infarct size, better cardiac function and viability compared to permanent occlusion. Multiparametric immunoimaging targeting CD11b+ cells by 89Zr-CD11b nanobody PET and Ly6Chi inflammatory monocytes by 64Cu-CCR2 PET demonstrated that mice subjected to transient coronary occlusion had less immune cell influx to the ischemic myocardium. This finding was confirmed by flow cytometry analysis of the infarct zone. In contrast, both myocardial infarction models cause a similar systemic immune response in the bone marrow and spleen as observed with multimodal imaging with subsequent similar numbers of CD11b+ cells in the blood. Both permanent and temporary coronary artery occlusion aggravate atherosclerosis in Apoe−/− mice with higher macrophage and Ly6Chi monocyte numbers in aortas and larger plaque size compared to Apoe−/− mice without myocardial infarction.

Conclusions

We developed and employed multimodal, multiparametric imaging protocols to characterize the immune response in the heart, bone marrow and spleen in two models of myocardial infarction. While cardiac function was superior in the ischemia reperfusion model, both types of myocardial infarction accelerated atherosclerosis.

Funding Acknowledgement

Type of funding sources: Other. Main funding source(s): NIH and DFG
A closed-chest model of selective atrial myocardial infarction for the study of induced electrophysiological and structural derangements
Authors: Amoros-Figueras G , Casabella-Ramon S , Company G , Arzamendi D , Macias Y , Jorge E , Sanchez-Quintana D , Rosell-Ferrer J , Guerra J M , Cinca J
Published: October 2022

Background

The diagnosis of atrial infarction is often masked by the frequent association with ventricular infarction. For this reason, the electrophysiological and structural consequences of selective atrial ischemia are poorly understood.

Purpose

The objective of this study was to analyze the alterations in the ECG, local electrograms, and multifrequential atrial myocardial impedance in a new experimental model of acute and chronic atrial infarction.

Methods

Seven anesthetized pigs were subjected to 4h of atrial ischemia induced by selective catheter occlusion of the atrial coronary branches originating in the left circumflex coronary artery. The surface ECG was recorded and the changes in P-wave morphology analyzed. Four weeks later the animals were subjected to endocardial voltage mapping (Carto) and multifrequential impedance. The hearts were processed for anatomopathological study.

Results

Selective occlusion of the coronary atrial branches induced atrial infarction with fibrosis in the left atrium in 6 of the 7 cases (Figure). The surface ECG showed prolongation of the P-wave duration (Figure) (P-wave in lead II: from 72±8ms at baseline vs. 97±18ms at 4 weeks, ANOVA p<0.01; P-wave in lead aVR: from 71±3ms at baseline vs 87±9ms at 4 weeks, ANOVA p<0.01) with no appreciable displacement of the PR segment. Endocardial mapping of the left atrium showed low-voltage bipolar zones with decreased multi-frequency impedance phase angle values, as compared with preserved zones of the same atrium (bipolar electrograms: from 0.6±1mV to 2.0±1.9mV, T-Test p<0.01; Phase angle at 300KHz: from −5.5±2° to −9.0±4.4° T-Test p<0.05).

Conclusion

We developed a closed-chest swine model of selective atrial infarction suitable for the study of ECG patterns and electrophysiological mechanisms linked to atrial myocardial ischemia and infarction. The structural derangements are detectable by endocardial mapping of local voltage electrograms and local tissue impedance.

Funding Acknowledgement

Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Supported by grants from ISCI-MINECO (FIS PI17/00069), FEDER, CIBERCV (CB16/11/00276)
Hypothermia yields favourable cardiac energetics in experimental ischemia/reperfusion – insights from non-invasive pressure-volume loops by CMR
Authors: Berg J , Jablonowski R , Nordlund D , Ryd D , Heiberg E , Carlsson M , Arheden H
Published: October 2022

Background/Introduction

In experimental studies, mild hypothermia (<35°C) has reduced infarct size [1], potentially through reduced reperfusion injury [2], but human trials have been ambiguous [3]. Furthermore, a pressure-volume (PV) loop framework is the gold standard for assessing cardiac performance, but the invasive approach limits its clinical use. Therefore, we used a porcine model of ischemia/reperfusion with cardiovascular magnetic resonance (CMR) imaging throughout one week to acquire non-invasive PV loops.

Purpose

To quantify the cardioprotective effects of hypothermia at reperfusion by serial CMR imaging in a perspective of cardiac energy and mechanics.

Methods

Normothermia and Hypothermia groups (n=7+7 pigs) were imaged with CMR at baseline and subjected to 40 minutes of normothermic ischemia by catheter intervention. Thereafter, the Hypothermia group was rapidly cooled (<35°C) for 5 minutes before reperfusion. Additional imaging at two hours, 24 hours, and seven days were used to assess ventricular volumes and ischemic injuries. Infarct size was determined by high-resolution ex-vivo T1-weighted images.

Results

Figure 1 shows that stroke volume (SV: p=0.029; Friedmans test) and ejection fraction (EF: p=0.068) were notably reduced post-reperfusion in the Normothermia group. In contrast, the decreases were ameliorated in the Hypothermia group (SV: p=0.77; EF: p=0.13). Mean arterial pressure (MAP) was stable in the Normothermia group (p=0.77) but dropped two hours post-reperfusion in hypothermic animals (p=0.007). Over one week, both groups experienced the same decrease and partial recovery pattern for stroke work, external power, and ventricular-arterial coupling. Still, the adverse effects from ischemia were generally attenuated in the Hypothermia group. Although not significant, ischemia/reperfusion caused smaller infarcts in the Hypothermia group (10±8% vs 15±8%; p=0.32). The change in PV loop variables from baseline to 24 hours was compared with infarct size for each group. Though statistical power was limited, the general trend in analyses of covariance indicated that hypothermia has cardioprotective properties incremental to reducing infarct sizes (Figure 2), such as higher external power (p=0.061) and lower arterial elastance (p=0.015).

Conclusion

Using non-invasive PV loops by CMR, we observed that mild hypothermia before reperfusion results in favourable cardiac energetics that alleviates the heart's work after an ischemia/reperfusion injury during the first week. This study suggests that hypothermia has cardioprotective properties, incremental to reducing infarct size. The primary cardioprotective mechanism was likely an afterload reduction unloading the injured left ventricle.

Funding Acknowledgement

Type of funding sources: Foundation. Main funding source(s): The Swedish Heart-Lung FoundationThe Medical Faculty of Lund University (ALF)
S1P lyase inhibition improves post-ischemic cardiac remodeling independently of infarct size via S1P receptor 1
Authors: Polzin A , Dannenberg L , Benckhoff M , Barcik M , Keul P , Weske S , Ahlbrecht S , Helten C , Haberkorn S , Floegel U , Zeus T , Mueller T , Graeler M , Kelm M , Levkau B
Published: October 2022

Background

Revascularization of the infarct vessel is the golden standard in acute myocardial infarction (AMI). Even in Western countries, more than ten percent of AMI patients are so-called “latecomers” and it is controversial if revascularization of the infarct vessel is beneficial in these patients. Current guidelines even discourage revascularization of the infarcted artery if symptom onset was >48 hours and the patient is asymptomatic. These patients are at high risk for cardiovascular events and heart failure with reduced ejection fraction (HFrEF). HFrEF has an enormous socio-economic impact, high morbidity, and mortality.

Purpose

Therapeutic options targeting post-ischemic cardiac remodeling are sparse. The bioactive sphingolipid sphingosine-1-phosphate (S1P) reduces ischemia/reperfusion injury when administered in advance. However, its impact on post-ischemic remodeling independently of its infarct size (IS)-reducing effect is yet unknown.

Methods

Acute myocardial infarction (AMI) in mice was induced by permanent ligation of the left anterior descending artery (LAD). C57Bl/6J were treated with the S1P lyase inhibitor 4-deoxypyridoxine (DOP) starting seven days prior to AMI to increase endogenous S1P concentrations. Cardiac function and myocardial healing were assessed by cardiovascular magnetic resonance imaging (cMRI), histomorphology and gene expression analysis. DOP effects were investigated in cardiomyocyte-specific S1P receptor 1 deficient (S1PR1 Cardio Cre+ and Cre− control) mice, and S1P concentrations measured by LC-MS/MS.

Results

S1P concentrations in plasma before induction of AMI were increased fourfold by DOP (Control 0.97±0.09μM [n=6] vs. DOP 3,80±0,09μM [n=6]). Scar size determined by MRI, as well as ejection fraction (EF), did not differ 24 h post AMI. In contrast, after 21 days, there was a clear difference between the two groups (scar size vehicle: 19.3±6.2% vs. DOP 13.4±5.7%; EF: Vehicle: 26.4±8.7% vs. DOP 38.2±11.8%). In addition, in the remote area 21 days post AMI in the DOP-treated animals, a reduced gene expression of brain natriuretic peptide, atrial natriuretic peptide and collagen 1a2. Finally, cardiomyocyte diameter in the remote myocardium was 21% smaller in DOP-treated (Vehicle: 21.95±1.59μm vs. DOP 17.35±0.77μm). The benefit of DOP-treatment was abolished in cardiomyocyte-specific S1PR1-deficient mice.

Conclusion

S1P improves cardiac function and myocardial healing post AMI independently of initial infarct size via S1PR1. Hence, in addition to its beneficial effects on I/R injury, S1PR1 may be a promising target in post-infarction myocardial remodeling as adjunctive therapy to revascularization and in patients who are not eligible for standard interventional procedures.

Funding Acknowledgement

Type of funding sources: None.