bluuebunny/crossref_metadata_2025_split huggingface dataset:
AT-RISK: AuToimmune disorders and cardiovascular RISK
Authors: Conrad N , Verbeke G , Molenberghs G , Goetschalckx L , Callender T , Rahimi K , Mason J C , McMurray J J V , Verbakel J Y 
Published: October 2022

Abstract

Background
Increased cardiovascular risk has been suggested for selected autoimmune disorders. Yet, for many conditions, evidence is insufficient to design more targeted prevention measures and guide clinical decisions.

Methods
We used linked primary and secondary electronic health records from the Clinical Practice Research Datalink (CPRD) to assemble a cohort of individuals newly diagnosed with any of 19 autoimmune disorders between 01/01/2000 and 31/12/2017 and free of cardiovascular disorders (CVD) up to 12 months after diagnosis, and up to five individuals matched on age, sex and socioeconomic status, with follow-up until 30/06/2019. We investigated incidence of nine cardiovascular outcomes and used cox-proportional hazards models to examine differences in patients with and without autoimmune disorders accounting for known cardiovascular risk factors.

Findings
We identified 432,883 individuals with autoimmune disorders and 2,159,350 matched controls. Of these, 104,504 people with and 450,195 without autoimmune disorders developed incident CVD during a median of 5.3 [interquartile range: 2.6, 9.6] years of follow-up. Patients with one or more autoimmune disorders had higher risk of cardiovascular outcomes: adjusted hazard ratio (HR) [95% confidence interval (CI)] 1.34 [1.31, 1.37]. This relationship held for every individual cardiovascular disorder and increased progressively with the number of autoimmune disorders present. Among autoimmune disorders, those associated with systemic inflammation, such as type I diabetes (2.09 [1.91, 2.28]), systemic sclerosis (2.90 [2.29, 3.67]) and systemic lupus erythematosus (2.21 [1.96, 2.50]) presented with highest overall cardiovascular risk (Figure).

Interpretation
Autoimmune disorders are associated with a high risk of developing cardiovascular outcomes. These findings warrant targeted cardiovascular prevention measures for patients with autoimmune disorders and further research into pathophysiological mechanisms underlying these complications.

Funding Acknowledgement
Type of funding sources: Public grant(s) – EU funding. Main funding source(s): This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie (grant agreement No 843267) and from the European Society of Cardiology (grant number App000037070).
Measurement error in echocardiographic assessment of aortic stenosis: an epidemiological consideration of research methodology and clinical practice
Authors: Velders B J J , Groenwold R H H , Kappetein A P , Braun J , Klautz R J M , Vriesendorp M D 
Published: October 2022

Abstract

Background/Introduction
Current international guidelines on treatment of valvular heart disease contain recommendations for aortic valve replacement based exclusively on abnormal echocardiographic parameters in asymptomatic patients with aortic stenosis (AS). To prevent misclassification and subsequent mistreatment of patients, these recommendations require accurate and unbiased measurements of hemodynamic parameters.

Purpose
To illustrate consideration and impact of measurement error in echocardiographic assessment of aortic stenosis in both the research and clinical setting.

Methods
First, a systematic review was performed to investigate the recognition of and correction for measurement error in clinical studies on the prognostic value of peak aortic jet velocity (Vmax), mean pressure gradient (MPG), and effective orifice area (EOA). Second, all potential erroneous sources in the calculation of those primary parameters were listed stratified to random or systematic measurement error, and subsequently their magnitude was quantified. Third, the impact of various types of measurement error on current thresholds for intervention was graphically illustrated in different clinical scenarios.

Results
The presence of measurement error was acknowledged in 44% of the 36 included studies, while none utilized methods to correct for it. Interobserver variability ranged between 0.9–8.3% for Vmax and MPG but was substantially higher for EOA (range 7.7–12.7%) implying lower reliability (Figure 1). Furthermore, the invalid assumption of a circular left ventricular outflow tract area resulted in a median underestimation in EOA of 22.5% compared to 3D-transesophageal echocardiography (3D-TEE), computed tomography (CT), and cardiovascular magnetic resonance (CMR) planimetry. Figure 2 illustrates the impact of this discrepancy on the classification of AS using one-sided t-tests to determine the areas under the curve; the proportion of patients with non-severe AS in a hypothetical cohort (based on values from the PARTNER 3 trial [1[) increased by 42%.

Conclusion(s)
Measurement error is underrecognized in studies of echocardiographic assessment of aortic stenosis. This review demonstrates that random and systematic measurement errors affect echocardiographic assessment, leading to potential misdiagnosis and subsequent mistreatment. Clinicians and scientists should be aware of the implications of measurement errors to enhance rightful clinical decision-making and assure research validity.

Funding Acknowledgement
Type of funding sources: None.
Medication adherence assessment and reporting in cardiovascular randomised controlled trials
Authors: Ashkir Z , Teoh Z , Ganesananthan S , Ahmed-Jushuf F , Beattie C J , Asher A , Kelshiker M , Howard J , Al-Lamee R 
Published: October 2022

Abstract

Background
Despite the knowledge that suboptimal adherence to medications may have an impact on the impact of treatment interventions, the assessment and reporting of medication adherence in cardiovascular randomised controlled trials has not been well studied. In this review we sought to study the differences in medication adherence assessment and reporting in cardiovascular randomised controlled trials (RCTs) comparing interventional procedures to medical therapy alone in four major cardiovascular conditions: coronary disease (CAD), atrial fibrillation (AF), heart failure (HF) and hypertension (HTN).

Methods
Comprehensive searches of PUBMED/MEDLINE, Clinicaltrials.gov and Cochrane Central Register of Controlled Trials (CENTRAL) were performed. Two independent authors screened and extracted general study data, adherence assessment methodology and adherence reporting characteristics from eligible RCTs.

Results
A total of 568 studies (257 HTN, 120 HF, 116 CAD and 75 AF RCTs) published between 2014–2019 were included in the review. Overall, only 44.7% of RCTs assessed adherence, 14.1% defined “good” adherence and 21.1% reported adherence results. HTN and CAD trials performed significantly better than HF and AF studies in all three parameters. Compared to drug trials, procedural trials were significantly less likely to assess adherence (21.1% vs 45.7%, p&lt;0.001), define good adherence (7% vs 15.5%, p&lt;0.001) or report adherence results (10.5% vs 21.3%, p=0.046). Adherence assessment practices were further significantly influenced by funding, placebo status and study outcome. Pill count/return of packaging was the most utilised method (49.5%) and direct observation the least utilised (1.1%). Adherence was calculated in 55.9% of RCTs with studies using different formulas and setting different thresholds for “good” adherence but most setting this at 80% adherence. Adherence reporting practices also varied significantly between RCTs. Most studies (53%) that assessed adherence, did not in fact report their results.

Conclusion
Our findings expose significant inconsistencies in adherence assessment and reporting practices amongst cardiovascular RCTs. This is a systemic problem with important implications on research quality and reliability. We therefore recommend the incorporation of mandatory adherence assessment and reporting into international reporting guidelines for RCTs such as the CONSORT statement. We also propose a grading system which once validated may be used to appraise standards of adherence assessment and reporting in RCTs.

Funding Acknowledgement
Type of funding sources: None.
Mechanism(s) for age-related sex differences in diet-induced cardiomyopathy: role of RNA methylation
Authors: Croft A , Kelly C , Chen D , Murtha L , Sugito S , Boyle A , Sverdlov A L , Ngo D T M 
Published: October 2022

Abstract

Background/Introduction
Age and sexual dimorphism contribute to the differential cardiometabolic dysfunctions associated with diet-induced obesity. However, the underlying mechanisms remain elusive. RNA modifications via RNA m6A methylation, is an emerging mediator of RNA stability, translation and localization responsible for regulation of multiple biological functions. RNA m6A modifications are regulated by “writers”, “erasers” and “readers”. The role of RNA methylation machinery genes in the heart is largely unexplored, and may provide insight into the influence of age and sex on cardiometabolic dysfunction.

Purpose
We aim to determine differential RNA methylation changes within the heart in a diet-induced cardiomyopathy mouse model, stratified according to age and sex.

Methods
Male and female C57BL/6 mice (6–8wk-old) were fed normal chow (NC) or high-fat/high-sucrose (HFHS) diet for 1 or 4 months. Echocardiographic measurements were performed at 1 and 4 months according to the American Society of Echocardiography and European Association of Cardiovascular Imaging guidelines. At study endpoint, glucose and insulin tolerance testing was conducted by injecting mice intraperitoneally with 2g/kg glucose or 0.6U/kg insulin, and monitoring blood glucose levels over a 2 hour period. RNA from heart tissue was subjected to quantitative PCR for RNA methylation machinery genes (FTO, ALKBH5, METTL3, METTL4, METTL14, YTHDF1 and YTHDF2). PPIA was used to normalise qPCR data.

Results
Both male and female mice showed evidence of age- and diet-induced metabolic dysfunction, however, males and females showed markedly different metabolic impairments. For example, glucose tolerance was exacerbated by 4 months of HFHS diet in males but not females; and only females showed impaired insulin tolerance. Echocardiography showed that males had systolic (stoke volume, cardiac output) and diastolic (E/A ratio) dysfunction after 4 months of HFHS diet, while females were unperturbed. We identified that METTL3 and METTL14, the “writers” of m6A methylation, were consistently increased in male mouse hearts at 1 vs 4 months of age but were unchanged or decreased in females, irrespective of diet. Conversely, YTHDF1, a “reader”, was unchanged in male mouse hearts at 1 vs. 4 months of age but was significantly increased in female hearts.

Conclusion
Our study confirms that markedly different cardiometabolic impairments occur in male versus female mice in response to long-term HFHS diet. Despite significant metabolic impairment in both sexes, cardiac dysfunction was only evident in males. RNA methylation machinery genes were differentially expressed in mouse hearts according to age and sex, suggesting that RNA methylation may be involved in age-related sexual dimorphism in cardiometabolic impairments.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): Heart Foundation
LTBP2 mutation may cause mitral valve prolapse
Authors: Durst R 
Published: October 2022

Abstract

Background
Mitral valve prolapse (MVP, MIM 157700) is a very common cardiac disorder occurring in 2.4% of the general population. MVP is characterized by myxomatous degeneration resulting in valve thickening, prolapsing and regurgitation. MVP results in mitral regurgitation, arrhythmias, bacterial endocarditis, congestive heart failure and sudden death. Familial studies of MVP suggest an autosomal dominant mode of inheritance with incomplete penetrance. We are wokring on defining the genetic causes of MVP.

Methods and results
We identified an MVP family with an LTBP-2 Val1506Met rare mutation (minor allele frequency of 0.0001) segregating with the MVP (LOD score of 1.5 out of a maximal potential of 2.6). LTBP2 is an excellent candidate gene for MVP because it regulates TGFβ signaling activity, shown to be disrupted in Marfan's syndrome mitral valvulopathy. High prevalence of MVP was reported in relatives of patients with Weill-Marchesani syndrome, caused by LTBP2 mutations.
We then generated two strains of mice using CRISPR technology. A complete knockout (deletion of the first exon) and a knock in mouse. Nineteen mice were dissected so far (figure 3). None of the wt mice (n=8) demonstrated myxomatous degeneration. Of the 11 mice with mutation, 8 homozygote for deletion and 3 homozygote for the human mutation, 8 demostrated myxomatous Degeneration by histology (1 knockin mouse ∼70% pentrance in male mice, p=0.001, Fisher exact test). The pathologist that read the studies was blinded to the genotype. Echocardiography of theses mice demonstrated findings consistant with MVP (deviation of the leaflets above the annluar line in parasternal long axis view) in 50% of affected mice and not in controls

Conclusion
We demonstrated that an LTBP2 mutation is segretating in a family with MVP. LTBP2 mutation mice model demonstrated myxomatous degeneration of the valve. Thus, LTBP2 may be an MVP causing gene

Funding Acknowledgement
Type of funding sources: Public hospital(s).
Demethylation and reactivation of the fibrosis-suppressor gene Rasal1 is initiated by R-loop formation which facilitates recruitment of TET3 and GADD45g
Authors: Maamari S , Xu X , Zeisberg E 
Published: October 2022

Abstract

Background
Silencing of fibrosis suppressor genes through DNA methylation has been shown to contribute to fibrosis progression in the heart and in other organs. Growth Arrest and DNA Damage 45-(GADD45) and Ten-Eleven-Translocation (TET) proteins are involved in active DNA demethylation, but how they are recruited to the promoter of fibrosis suppressor genes has not yet been understood.

Purpose
To identify the mechanisms involved in site-specific demethylation of the promoter region of the cardiac fibrosis suppressor gene Rasal1.

Methods
We previously identified lncRNA Gm15749 located within the Rasal1 promoter region. Using DRIP qRT-PCR, we demonstrated an enrichment of R-loops at the Rasal1 promoter in non-fibrotic mouse cardiac fibroblasts. Such enrichment of R-loops was lost upon TGF-beta-treatment in activated “fibrotic” fibroblasts, and also upon Gm15749 knockdown in untreated non-fibrotic fibroblasts. ChIP qRT-PCR further revealed binding of GADD45g to the Rasal1 promoter and Co-IP experiments confirmed a physical interaction of TET3 and GADD45g.

Results
Studying the RAS Protein Activator Like 1 (Rasal1), which has been shown to be silenced in fibrosis of the heart due to promoter hypermethylation, we find that the lncRNA Gm15749 within the promoter region of Rasal1 is protective by forming an R-loop at the Rasal1 promoter. Gm15749 mediates the recruitment and binding of GADD45g, which then triggers the demethylation process by recruiting TET3 specifically to the Rasal1 promoter region. During cardiac fibrosis, all three factors; Gm15749, TET3, and GADD45g are significantly downregulated and the protection of Rasal1 is lost due to the failure of the formation of the demethylation complex.

Conclusions
Our results suggest a complex involving an R-loop forming lncRNA termed Gm15749, which interacts with the demethylation complex consisting of GADD45g and TET3 to perform site-specific demethylation at the Rasal1 promoter. Such R-loop formation may counteract fibrogenesis through reactivation of fibrosis-suppressor genes silenced by hypermethylation.

Funding Acknowledgement
Type of funding sources: None.
A large-scale functional high-throughput screening identifies miR-515 and miR-519e as potent inducers of human iPSC-cardiomyocyte proliferation
Authors: Renikunta H V , Lazarow K , Gong Y , Shukla P C , Giral H , Kratzer A , Nageswaran V , Opitz L , Engel F B , Haghikia A , Paneni F , Von Kries J P , Streckfuss-Boemeke K , Landmesser U , Jakob P 
Published: October 2022

Abstract

Introduction
Ischemic heart failure persists as a global health problem despite optimized medical and adjunctive device therapies. Loss of cardiomyocytes in the absence of a proliferative response comprise a major contributor to pathological remodeling and death in this patient population. Experimental studies have shown that microRNAs (miRNAs) may be used as a therapeutic option to reinduce adult cardiomyocyte proliferation.

Purpose
This study thought to evaluate proliferative potential in human cardiomyocytes after overexpression and inhibition of 2019 miRNAs.

Methods
To identify miRNAs that regulate cardiomyocyte proliferation, we performed functional high-throughput screenings in human iPSC-derived cardiomyocytes (hiPSC-CM) after transient hypoxia. Herein, 2019 miRNA-mimics for overexpression and 2019 anti-miRs for inhibition were individually transfected to examine EdU-incorporation in hiPSC-CM. MiR-mimic-515 and miR-mimic-519e that induced the highest EdU-uptake, were further assessed by immunostaining and molecular methods for markers indicative of early and late mitosis. In addition, RNA-Sequencing in hiPSC-CM after overexpression of miR-515 and miR-519e was performed to examine differential gene expression and miRNA-modulated pathways involved in cardiomyocyte proliferation.

Results
Using a functional high-throughput screening, we assessed differential proliferative potential of 2019 miRNAs after transient hypoxia by transfecting both miR-inhibitor and miR-mimic libraries in human iPSC-derived cardiomyocytes (hiPSC-CM). Overexpression of 28 miRNAs substantially induced proliferative activity in hiPSC-CM, with an overrepresentation of miRNAs belonging to the C19MC-cluster and adjacent miR-371–373 family. Two of these miRNAs, miR-515 and miR-519e increased markers of early and late mitosis, with an additive cardiomyocyte turnover after transient hypoxia and substantially increased Aurora B-kinase activity in midbodies, indicative of cell division. These findings were supported by molecular studies using qRT-PCR, Western blot, and RNA-Sequencing after overexpression of miR-515 and miR-519e showing substantial alterations of signaling pathways relevant for cardiomyocytes proliferation in human iPSC-CM.

Conclusion
Collectively, these results support a critical role of miR-515 and miR-519e for induction of proliferation in human cardiomyocytes under hypoxic conditions, such as present in patients with ischemia-driven cardiomyopathy.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): This work was supported by the German Centre for Cardiovascular Research (DZHK), Deutsche Stiftung für Herzforschung (DSHF) and OPO Foundation.
Chronic insulin resistance deteriorates cardiac mitochondrial quality control in a mouse model of SEC-NAFLD-IR
Authors: Lu K , Rothe M , Floegel U , Kotzka J , Oehler D , Roden M , Kelm M , Szendroedi J , Westenfeld R 
Published: October 2022

Abstract

Background
Insulin resistance and nonalcoholic fatty liver disease (NAFLD) both relate to cardiovascular mortality. Using a mouse model of chronic lipid overload and secondary-NAFLD-induced insulin resistance (SEC-NAFLD-IR), we recently deciphered that SEC-NAFLD-IR already at young age provoked myocardial lipotoxicity with reduced mitochondrial efficiency and increased vulnerability to cardiac ischemia. However, long-term consequences of SEC-NAFLD-IR remain elusive.

Purpose
Here we aimed to elucidate the impact of long-term SEC-NAFLD-IR on multiple mitochondrial quality control (mQC) mechanisms in the heart and its consequences for cardiac function.

Methods
We studied 36 SEC-NAFLD-IR mice (72-week-old). For mechanistic experiments, we applied palmitate-induced insulin resistant murine HL-1 cells. Cardiac mitochondrial dynamics were measured via quantification of mitochondrial morphology and expression of mitochondrial fusion and fission factors (Opa1, Drp1, Fis1, Mfn 1 & 2). Mitophagy level was evaluated via immunofluorescence and protein expression of key mitophagy-related genes (Parkin, NIX, LC3). Mitochondrial biogenesis and mass were examined via quantitation of PGC-1α expression, mtDNA and citrate synthase activity.

Results
72-week-old SEC-NAFLD-IR mice exhibited 21% (p=0.001) and 32% (p&lt;0.001) higher body weight and heart weight compared with controls. Along with elevated oxidative stress, hepatic lipid accumulation and inflammation, 6h-fasted SEC-NAFLD-IR mice were characterized by increased plasma glucose, insulin and cholesterol. SEC-NAFLD-IR mice displayed a cardiac phenotype with 21% higher left ventricular mass (normalized to body weight, p&lt;0.001) and 6% lower ejection fraction compared to controls (73.5% SEM 0.90 vs 69.4% SEM 1.65, p=0.04). We found several advantageous mQC mechanisms suppressed in aged SEC-NAFLD-IR mice including long form OPA1-mediated mitochondrial fusion, Parkin- and NIX-mediated mitophagy. Likewise, mitochondrial biogenesis was suppressed in the aged insulin-resistant heart, which was connected to a 65% downregulation of PGC-1α1 expression (p=0.01). Interestingly, downregulation of cardiac PGC-1α1 in aged SEC-NAFLD-IR mice coincided with upregulation of PARIS, indicating the crucial participation of the Parkin/PARIS pathway in mQC of the insulin-resistant heart. In addition, induction of insulin resistance in murine HL-1 cardiomyocytes also led to increased mitochondrial fragmentation and decreased PGC-1α1 expression.

Conclusion
This study demonstrated that regulation of mitochondrial network and turnover is hampered by SEC-NAFLD-IR in the hearts of aged mice, which may contribute to hypertrophy and cardiac dysfunction in insulin resistance.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Collaborative Research Centre 1116 (German Research Foundation)