bluuebunny/crossref_metadata_2025_split huggingface dataset:
Extended follow-up data: impact of selective loss to follow-up in a trial of secondary prevention of coronary artery disease
Authors: Tijssen A , Snaterse M , Jorstad H.T , Scholte Op Reimer W.J.M , Peters R.J.G 
Published: November 2020

Abstract

Background
We performed an extended follow-up of the RESPONSE-2 trial, which evaluated a new secondary prevention strategy for patients with coronary artery disease (CAD). Selective loss to follow-up (LTFU) however, may induce biased results.

Objectives
To assess LTFU and its implications on the internal validity of the trial at extended follow-up.

Methods
RESPONSE-2 (N=824) was a randomised trial of nurse-coordinated referral of CAD patients to ≤3 commercial lifestyle interventions, on top of usual-care, or to usual-care alone. The primary outcome was assessed at 1-year follow-up, after which the interventions were discontinued. An extended follow-up was performed two years later. We evaluated overall selective LTFU, between-group selective LTFU and within-group selective LTFU, by comparing baseline characteristics, compliance with the lifestyle interventions (intervention group only) and achievement of lifestyle-related risk factor (LRF) targets at 1-year follow-up for attendees and non-attendees of the extended follow-up. In all variables a standardized mean difference (SMD) of ≥0.2 with a p-value &lt;0.05, based on an independent t-test for continuous data and fisher's test for discrete data, was considered evidence of selective LTFU.

Results
In total, 520 patients attended the extended follow-up: 266/411 in the intervention and 254/413 in the control group. Patients LTFU were generally younger (56.5 vs. 59.3 years; SMD=0.299), more often baseline smokers (37% vs. 22%; SMD=0.334) and had less often quit smoking directly after hospitalization (17% vs. 25%; SMD=0.203). We found no evidence of between-group selective LTFU and several indications of within-group selective LTFU (table). Intervention group attendees were more compliant with the lifestyle interventions compared with non-attendees (weight loss: SMD=0.450; p≤0.05 and physical activity: SMD=0.516; p≤0.01)

Conclusion
Baseline LRF control, compliance with the lifestyle interventions and achievement of LRF targets were less favourable for non-attendees of the extended follow-up. Effective handling of missing data is recommended to minimize loss of contrast due to within-group selective LTFU and to ensure generalizability of results.

Funding Acknowledgement
Type of funding source: Private company. Main funding source(s): The RESPONSE-2 trial was sponsored by Weight Watchers International, Inc. (New York, New York) and Philips Consumer Lifestyle (the Netherlands). The sponsors had no role in the design, data collection, data analysis, data interpretation, and writing of the manuscript.
The prevalence and patient outcomes of adult primary hypercholesterolaemia and dyslipidaemia in the UK: a longitudinal retrospective study using a primary care dataset from 2008 to 2018
Authors: Bilitou A , Rabe A , Inema L , Alamgir G , Dunton K 
Published: November 2020

Abstract

Background
Cardiovascular disease (CVD) remains the leading cause of death and morbidity in Europe and United Kingdom (UK). Increased low-density lipoprotein cholesterol (LDL-C), implicated in primary hypercholesterolaemia and mixed dyslipidaemia (PH/MD), is an extensively studied risk factor, with proven direct and linear causality of CVD. Lowering LDL-C remains a primary goal in the treatment and prevention of atherosclerotic CVD.

Objectives
This study aimed to quantify adult prevalence and incidence of PH/MD in the UK.

Methods
We used an anonymised dataset covering primary care practices across the UK, accessed through the Clinical Practice Research Datalink (CPRD GOLD). Ethics approval was sought and provided (Protocol 19_238R). Using a validated set of clinical codes as well as pre-treatment lipid profile levels for total cholesterol (TC) (&gt;8 mmol/L), LDL-C (&gt;4.9 mmol/L), we calculated prevalent and new adult cases starting from 2009 to 2018. Our denominator population was the CPRD GOLD GP-registered adult population each calendar year, adjusted for mortality.

Results
There were 1,514,916 adults in the CPRD GOLD GP register for the period from 2009 to 2018. During that period there were 354,444 patients diagnosed with PH/MD. Males comprised 46.5%, mean age on diagnosis was 58.2 years. Mean follow-up time was 104 months. The annual prevalence of PH/MD increased from 2009 to 2019 (see graph). The overall prevalence across the period was 22.7%. Mean annual incidence across the decade was 1.7%. Mean LDL-C levels were 4.7 mmol/L and mean TC level was 6.8 mmol/L. In this cohort, 15.9% experienced cardiovascular events (see table). Nearly all patients have used lipid lowering therapies. Only 2.2% achieved at least 40% reduction of LDL-C from baseline.

Conclusions
The prevalence of PH/MD has been increasing despite the availability of interventions. Considering guidelines, only a small proportion of patients have achieved LDL-C goals.
Est prevalence of PH/MD in the UK

Funding Acknowledgement
Type of funding source: Private company. Main funding source(s): Daiichi Sankyo Europe, Health iQ Ltd
Effect of PM2.5 for hospitalization of cardiovascular diseases and medical expenses by age group: a nationwide study from the Japanese Registry Of All cardiac and vascular Diseases (JROAD)
Authors: Kaihara T , Yoneyama K , Kuwata S , Mitarai T , Watanabe M , Koga M , Kamijima R , Ishibashi Y , Izumo M , Tanabe Y , Higuma T , Harada T , Akashi Y.J 
Published: November 2020

Abstract

Introduction
Although particulate matter with an aerodynamic diameter less than or equal to 2.5 μm (PM2.5) exposure influences cardiovascular events, there is limited knowledge how PM2.5 exposure is associated with cardiovascular hospitalization in Japan.

Purpose
We tested the hypothesis that PM2.5 exposure was related to the number of cardiovascular hospitalizations, hospitalization period, and medical expenses.

Methods
We included 835,405 subjects who were admitted to acute care hospitals in Japan. We classified PM2.5 exposure concentration into quintile groups (from low to high in the order of “PM-1” to “PM-5” group). Multilevel mixed-effects Poisson and linear regression analysis were used to estimate the association of PM2.5 exposure concentration with the cardiovascular hospitalization events, duration and medical expenses. We also analyzed those factors classified by age.

Results
PM-2, 3, 4, 5 group were positively related to the number of cardiovascular hospitalization events compared with PM-1 group (Table 1). PM-3, 4, 5 group were positively associated with a 0.018, 0.029, and 0.029 (all p&lt;0.001) of the difference of log cardiovascular hospitalization period compared with PM-1 group (p for trend &lt;0.001). These groups were also positively associated with a 0.016, 0.023, and 0.021 (all p&lt;0.001) of the difference of log medical expenses compared with PM-1 group (p for trend &lt;0.001). Analyzed by age group, hospitalization duration was longer (p&lt;0.05) and medical expenses was lower (p&lt;0.05) significantly in super-aging group (over 90 years old) than the group under 64 years old (Figure 1).

Conclusions
PM2.5 exposure concentration has harmful effect on not only cardiovascular events but cardiovascular hospitalization period and medical expenses due to big-data in Japan. Medical costs were kept low in comparison with their hospitalization period in subjects over 90 years old.
Figure 1

Funding Acknowledgement
Type of funding source: None
Single-cell RNA sequencing of human fetal epicardium reveals novel markers and regulators of EMT
Authors: Streef T.J , Van Herwaarden T , Smits A.M , Goumans M.J 
Published: November 2020

Abstract

Background
The heart is covered by the epicardium, consisting of epithelial cells and a mesenchymal layer. The epicardium has been shown to be essential during cardiac development by contributing cells through epithelial-to-mesenchymal transition (EMT) and the secretion of paracrine factors. In the adult, the epicardium conveys a cardioprotective response after myocardial infarction, albeit suboptimal compared to the epicardial contribution to heart development. Although the developing epicardium has been characterised in mice and zebrafish, knowledge on the human fetal epicardium derives mostly from cell culture models. Therefore, direct analysis of the human fetal epicardium is vital as it provides new insights into the cellular and biochemical interactions within the developing heart, which can potentially contribute to enhancing the post-injury response.

Aim
To study the human fetal epicardium using single-cell RNA sequencing (scRNA seq) in order to determine its cellular compositionThe data are further explored to e.g.identify regulators of epicardial EMT.

Methods
Epicardial layers were isolated from four fetal human hearts (14–15 weeks gestation, obtained under informed consent and according to local ethical approval). Tissue was digested, and single live cells were sorted into 384-wells plates and sequenced. Data analysis was performed using R-packages RaceID3 and StemID2. Findings were validated using qPCR and immunohistochemistry.

Results
Analysis of 2024 cells reveals a clear clustering of the epicardial epithelium and the mesenchymal population. Importantly, we found that “classical” markers, such as Wilms' Tumor 1 and T-box transcription factor 18, are not specific enough to reliably identify the epicardium, but our analysis has provided markers that do allow for robust identification of the epicardium. Additionally, we were able to identify epicardial subpopulations based on their expression profile, and we are currently investigating these using immunohistochemistry in human fetal and adult heart tissue sections. To establish the regulation of epicardial activation we are focussing on the process of EMT within our dataset using RaceID2. From our analysis, several regulators of epicardial EMT are proposed that will be followed up on in vitro.

Conclusions
We identify various novel markers of the fetal epithelial epicardium, as well as characterizing markers of the mesenchymal layer. We also identified novel factors involved in epicardial EMT, and these are currently being validated in our cell-culture model. These data can provide new insights into the post-injury response in the adult heart.

Funding Acknowledgement
Type of funding source: Public Institution(s). Main funding source(s): Dutch Heart Foundation
VENTHEART is required for cardiomyocyte specification and function
Authors: Jonsson Boezelman M , Dashi A , Tan W , Pan B , Ilmari A.M , Tiang Z , Hartman R.J.G , Anen C.G.O , Liu Z , Wu J.J , Bin G , Lim B , Walentinsson A , Foo R.S 
Published: November 2020

Abstract

Background
Long noncoding RNAs (lncRNAs) control early stages of cardiac differentiation, however their role in later specification and maturation is still not well explored.

Methods and results
We performed single cell RNA-seq for 2, 6 and 12 week-old hESC-CM. Weighted correlation network analysis (WGCNA) identified core genes significantly upregulated, along with a subset of lncRNAs. Importantly, these lncRNAs are highly abundant and unique to human heart. Through independent integrative analysis of genome-wide association studies (GWAS) and expression quantitative trait locus (eQTL) data using human hearts, we also identified a long intergenic noncoding RNA (we call VENTHEART, VHRT) as co-regulated with core cardiac contractile genes, and strongly associated with heart failure. VHRT was highly expressed in MYL2+ hESC-CMs in our single cell dataset, and its locus is antisense and downstream of MYL2. VHRT knockdown (KD) in 6-weeks old hESC-CMs downregulated MYL2 and other key cardiac genes. Patch clamp recordings with VHRT KD cells showed a loss of the ventricular-like action potential. Concordantly, CRISPR-mediated excision of the VHRT locus led to impaired CM sarcomere formation, and loss of CM specification gene programs. VHRT transcript replacement in VHRT-KO cells was however insufficient to rescue the phenotype. Instead, we established by 3C assay, that the VHRT locus loops and interacts with the MYL2 promoter, bearing histone marks characteristic of a super-enhancer.

Conclusion
Thus, we conclude that both the VHRT lncRNA transcript and its genomic locus are required for proper CM specification and function, and may play a role in heart failure progression.

Funding Acknowledgement
Type of funding source: Other. Main funding source(s): EMBO, Singapore National Research Council
Influence of perfusable microvasculature on excitation-contraction coupling in IPSC-derived myocardium
Authors: King O , Cruz-Moreira D , Kit-Anan W , Sayed A , Wang B.X , Fourre J , Randi A.M , Rasponi M , Terracciano C.M 
Published: November 2020

Abstract

The myocardium is one of the most densely vascularised tissues in the body, with dynamic metabolic demand from beating cardiomyocytes (CM) necessitating an intimate relationship with microvasculature. Endothelial cells (EC) produce a diverse array of cardio-active factors which acutely and chronically modulate myocardial phenotype. Disruption of CM-EC signalling results in pathological remodelling, and ultimately organ failure. However, as physiologically relevant recapitulation of CM-EC interaction has been difficult to achieve in vitro, many molecular mechanisms governing their interaction remain poorly understood.
To induce cardiac vasculogenesis in vitro, we have developed microfluidic chips which subject 3D hydrogel cultures to precisely controlled flow. We then co-cultured human cardiac microvascular ECs, human left ventricular fibroblasts (FB), and human induced pluripotent stem cell-derived cardiomyocytes for 5 days under a pro-vasculogenic protocol (0.5 ul/min flow rate, 50ng/ml VEGF, 100ng/ml Ang-1). Via live and fixed immunofluorescence microscopy, we observed spontaneous formation of a microvasculature network with a continuously open lumen embedded within beating myocardium. Simultaneous quantification of iPSC-CM contractility and perfused red blood cell velocity reveals biomimetic pulsatile flow profile within the microvasculature. To evaluate the influence of microvasculature on CM function, we incorporated CMs differentiated from stem cells with the genetically encoded calcium biosensor GCaMP6F. Compared to CM only control, vascularised preparations demonstrate significantly faster calcium transient time to peak (−11.5%, p=0.007) and time to 50% relaxation (−15%, p=0.01). Under static conditions and 1Hz electrical stimulation, presence of EC was associated with reduced iPSC-CM arrhythmia at baseline (p&lt;0.0001) and during 1uM isoprenaline treatment (p=0.0003), while maintaining isoprenaline induced Ca2+ handling quickening.
To the best of our knowledge, this work represents the first fully perfusable model of the myocardial microvasculature, and highlights the importance of EC regulatory influence on CM function. Further work aims to investigate underlying molecular mechanisms to provide therapeutically relevant insight into cardiac biology.

Funding Acknowledgement
Type of funding source: Foundation. Main funding source(s): British Heart Foundation
Cholinergic nerve regulation of heart regeneration
Authors: Mahmoud A 
Published: November 2020

Abstract

Background
Cardiac nerves regulate many important physiological functions of the heart such as heart rate and contractility. The emerging role of cardiac nerves during tissue homeostasis and regeneration is beginning to be appreciated. We discovered that neonatal mice are capable of regenerating their hearts following injury within a brief period after birth by proliferation of the pre-existing cardiomyocytes. Furthermore, we have demonstrated that cholinergic nerves play an important role in guiding the neonatal heart regenerative response. However, the adult mammalian heart, including the human heart, is incapable of regeneration following injury. Thus, there is great excitement about understanding the evolutionarily conserved mechanisms of endogenous cardiac regeneration, so that we can explore potential avenues to reawaken this process in adult humans.

Purpose
Our overarching goal is to define the mechanisms by which cholinergic nerves regulate heart regeneration following ischemic injury by using the neonatal mouse heart regeneration model. These studies will uncover novel pathways by which cholinergic signaling promotes cardiomyocyte proliferation and heart regeneration, which holds significant therapeutic potential for treatment of adult heart disease.

Methods
In this project, we employed genetically engineered mouse models of the critical receptors for cholinergic signaling in the heart to define the mechanisms of cholinergic nerve regulation of heart regeneration. First, we generated a cardiomyocyte-specific deletion of the muscarinic receptor (M2), the most predominant muscarinic receptor subtype present in the heart. In addition, we utilized the α7 nicotinic receptor (Chrna7) knockout mice to study the role of Chrna7 in endogenous immune cells, which is the main mediator of the cholinergic anti-inflammatory pathway. These mouse models will address how cholinergic nerves regulate heart regeneration via the M2 muscarinic receptor signaling and the inflammatory response following injury.

Results
Our results demonstrate that inhibition of two different cholinergic receptors (muscarinic and nicotinic) results in a reduction in cardiomyocyte proliferation and inhibition of the neonatal cardiac regenerative response following injury. More importantly, we demonstrate that cholinergic signaling mediates the cardiac regenerative response mainly through suppression of pro-inflammatory cytokines via the cholinergic anti-inflammatory pathway.

Conclusions
Cholinergic nerve signaling plays an important role in mounting a robust cardiac regenerative response following injury. These results have significant therapeutic potential, which will forge new paradigms with respect to the role of cardiac nerves during mammalian cardiac regeneration and reveal potential mechanisms regarding the benefits of nerve stimulation following cardiac injury in humans.

Funding Acknowledgement
Type of funding source: Foundation. Main funding source(s): American Heart Association, Wisconsin Partnership Program
ADAM10 inhibition improves survival and augments cardiac function after myocardial infarction
Authors: Klapproth E , Kuenzel S , Guenscht M , Lorenz K , Weber S , Guan K , El-Armouche A 
Published: November 2020

Abstract

Background and purpose
Following myocardial infarction (MI), adverse fibrotic remodeling with extensive deposition of extracellular matrix (ECM) components has substantial consequences for the contractility of the ventricle finally leading to terminal heart failure (HF). Recently, inhibition of ECM-remodeling enzymes is discussed as potential treatment option for HF, especially following MI. The metalloprotease ADAM10 plays a crucial role in the development of the cardiovascular system and HF patients show elevated serum levels of the ADAM10 substrates CXCL16 and FasL. However, the causal role of ADAM10 in cardiovascular diseases has not been investigated. Here we evaluate the so far unknown role of ADAM10 in heart failure and after MI.

Methods and results
Our study capitalized from human atrial tissue biopsies, a cardiomyocyte-specific ADAM10 knockout (ADAM10 KO) mouse model as well as pharmacological ADAM10 inhibition following MI. ADAM10 expression analysis revealed elevated protein levels in HF patients compared to non-failing hearts. Upon MI, ADAM10 KO and pharmacological ADAM10 inhibition (GI254023X) significantly improved overall survival, significantly enhanced cardiac function (fractional area shortening - FAS, ejection fraction - EF) and significantly reduced infarct sizes. Compared to the high potential angiotensin receptor neprilysin inhibitor (ARNi) LCZ696, ADAM10 inhibition and combined ADAM10i/LCZ696 treatment resulted in preservation of cardiac function that was superior to sole LCZ696 treatment. Mechanistically, this functional improvement was due to reduced shedding of the ADAM10 substrate Notch1, induction of angiogenesis and an ADAM10-dependend inactivation of the NLRP3 inflammasome

Conclusion
Our data suggest that ADAM10 targeting is highly efficient for improving post-infarction cardiac function. Due to its overexpression in heart tissue of HF patients, ADAM10 could be a potential molecular target to improve therapy after MI. In terms of overall survival and pathophysiological remodeling following MI, our data suggest a greater potential of the ADAM10i/LCZ696 combinatorial therapy than sole LCZ696 treatment.

Funding Acknowledgement
Type of funding source: Public grant(s) – National budget only. Main funding source(s): German Heart Foundation/ German Foundation of Heart Research
Protective action of the microbial metabolite butyrate against cardiomyocyte hypertrophy
Authors: Umei M , Akazawa H , Saga-Kamo A , Yagi H , Liu Q , Kadowaki H , Matsuoka R , Matsunaga H , Nomura S , Komuro I 
Published: November 2020

Abstract

Introduction
Short-chain fatty acids are one of the gut microbial metabolites that may influence host physiology. We previously reported that gut dysbiosis was associated with heart failure, and that the proportions of butyrate-producing bacteria diminished prominently in the gut of patients with heart failure.

Purpose
We investigated the molecular mechanism of butyrate and investigated the protective mechanism against heart failure.

Methods
We searched for G protein-coupled receptors for short-chain fatty acids using single-cell transcriptome analysis of cardiomyocytes and non-cardiomyocytes isolated from murine hearts. In addition, we examined the effects of butyrate on endothelin-1 (ET1) or isoproterenol-induced hypertrophic responses and histone deacetylase (HDAC) activities in cultured neonatal rat cardiomyocytes.

Results
Single-cell transcriptome analysis and co-expression network analysis revealed that G protein-coupled receptors for short-chain fatty acid receptors were not expressed in cardiomyocytes and that Olfr78 was expressed in vascular smooth muscle cells in the heart. Treatment with butyrate inhibited ET1-induced hypertrophic growth and up-regulation of the genes such as Nppa, Acta1, and Myh7 in cultured rat neonatal cardiomyocytes. Moreover, butyrate increased the acetylation levels of histone H3, indicating that butyrate has an inhibitory effect on HDAC in cardiomyocytes. In addition, treatment with butyrate caused up-regulation of Inpp5f, encoding inositol polyphosphate-5-phosphatase f, which was associated with a significant decrease in the phosphorylation levels of Akt. These results suggest that butyrate may act as HDAC inhibitor to increase Inpp5f gene expression, leading to the activation of Akt-glycogen synthase kinase 3beta (Gsk3beta) pathway, and thereby protect against hypertrophic responses.

Conclusion
There was no known GPCR for short-chain fatty acid expressed in cardiomyocytes. However, butyrate suppressed cardiomyocyte hypertrophy through epigenetic modification of gene expression. Our results may uncover a potential role of the dysbiosis of intestinal microbiota in the pathogenesis of cardiac hypertrophy and failure.

Funding Acknowledgement
Type of funding source: None