bluuebunny/crossref_metadata_2025_split huggingface dataset:
Doxorubicin-induced upregulation of follistatin-like 3 (FSTL3): a new therapeutic target
Authors: Chen D , Croft A , Kelly C , Haw T J , Leong A , Sverdlov A , Ngo D 
Published: October 2022

Abstract

Introduction
Doxorubicin (DOX) is among the most used anticancer drugs with associated cardiotoxicity. Follistatin-like 3 (FSTL3), a secreted member of the follistatins family that can selectively bind to members of the TGF-β superfamily, is involved in regulation of cardiac hypertrophy and heart failure. FSTL3 is also upregulated in breast and colorectal cancer tumours, is also an unfavourable prognostic indicator for various cancers.

Purpose
We aim to determine the dual role of FSTL3 in prevention of DOX-induced cardiotoxicity and synergistic anti-cancer effects.

Methods
Human cardiomyocytes (HCMs) were treated with DOX at 1uM (EC50) for 72 hours. Cell viability was assessed via CellTiter-Glo®. Secreted FSTL3 levels, as measured by ELISA (R&D systems). FSTL3 and TGF-β mRNA levels were measured by qPCR. Co-treatment of DOX with human anti-FSTL3 antibodies (Aviva Systems Biology) at 10ug/mL were introduced for 72hrs treatment.

Results
Secreted FSTL3 levels were significantly increased in DOX-treated HCMs at 72hrs compared to control (n=5, p&lt;0.001). Consistently, FSTL3 and TGF-β mRNA levels, in collected HCMs were significantly increased in DOX-treated cells. Co-treatment of DOX with human anti-FSTL3 antibodies at 10ug/mL significantly improved HCM viability compared to IgG control group. Conversely, anti-FSTL3 antibodies provided synergistic anti-cancer effects with DOX: MCF-7 breast cancer cells were significantly reduced when co-treated with DOX and anti-FSTL3 antibody vs. IgG controls.

Conclusion
We show, for the first time, that: 1) FSTL3 is secreted directly from HCMs; 2) FSTL3 levels (both circulating and mRNA) is markedly elevated with DOX treatment; 3) neutralisation of FSTL3 in DOX-treated HCMs, restored HCM viability; and 4) exhibit synergistic anti-cancer effects with DOX. Taken together, FSTL3 is a potential target for dual anti-cancer and cardioprotective effects.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Heart Foundation of Australia Future Leader FellowshipsNSW Ministry of Health EMC FellowshipNSW Ministry of Health Translational Research Grant
Nucleic acid-based therapies using living myocardial slices
Authors: Nunez-Toldra R , Del Canizo A , Secco I , Nicastro L , Giacca M , Terracciano C M 
Published: October 2022

Abstract

Background
Gene therapy based on viral vectors is a promising new approach and offers great potential for the study and the treatment of cardiac diseases. Nucleic acid-based transfection of human heart tissue could be considered a valid and translational alternative to the use of transgenic animal models in cardiovascular research.

Purpose
Here we explore the use of Living Myocardial Slices (LMS) as a platform for nucleic acid-based therapies. Rat LMS and Adeno-Associated viruses (AAV) were used to optimise and analyse gene transfer efficiency, viability, tissue functionality, and cell tropism in cardiac tissue. Human cardiac samples from failing (dilated cardiomyopathy) hearts were also used to validate the human model. In addition, we transfected and tested for the first time the regenerative effect of AAV-miR-199a in both healthy and pathologically overloaded LMS.

Methods
LMS (300 μm thick) were prepared and glued to PTFE-coated holders, mounted on custom stretchers, and cultured at physiological sarcomere length for 72-hours under electrical stimulation. Two recombinant AAV serotypes (AAV-6 and AAV-9) at different multiplicity of infection (MOI) expressing green fluorescent protein (GFP) or the miR-199 transgene were added to the surface of the LMS.

Results
AAV6 proved to be the most suitable serotype and 20,000 MOI the most efficient dose, as these conditions were proven not to affect LMS contractility and to have the highest transduction and penetrability efficiency. This serotype exhibited preferential cell tropism to cardiomyocytes and stromal cells (40% of transduction in both) in rat LMS, with lower efficiency in endothelial cells, (20% of transduction). In contrast, AAV6 in human failing LMS showed higher transduction efficiency in endothelial cells and myofibroblasts. Treatment with miR-199a significantly increased the active force of pathologically overstretched LMS, downregulated the expression of its target genes, and affected the proliferative capacity of LMS.

Conclusion
LMS can be used as a model for nucleic acid-based therapies and their transfection protocol can be adapted to samples obtained from failing human hearts. With great translational relevance, this model would accelerate preclinical studies of novel nucleic acid-based therapies for heart failure.

Funding Acknowledgement
Type of funding sources: Foundation. Main funding source(s): British Heart Foundation
Autoantibodies against the cardiovascular protective BPIFB4 in hospitalized patients with COVID-19
Authors: Ciaglia E , Montella F , Lopardo V , Esposito R M , Guarracino F , Spinetti G , Maciag A A , Ciccarelli M , Vitale C , Pellegrino S G , Polverino B , Izzo C , Vecchione C , Puca A A 
Published: October 2022

Abstract

Background/Introduction
The bactericidal/permeability-increasing fold-containing family-B-member-4 (BPIFB4) serves as a biomarker of healthy aging [1,2] and displays prognostic relevance in vascular pathology [3–5]. We recently described a drop in plasma BPIFB4 level in patients with severe COVID-19 compared to low-grade disease patients [6].

Purpose
As COVID-19 is associated with autoimmune features, we developed the methods for determination of Anti-BPIFB4 IgG (autoAbs) and then characterized their neutralizing activity in COVID-19 patients.

Methods
A sandwich ELISA-based colorimetric assay followed by immunoblot analysis detected the presence of autoAbs against BPIFB4 in 60 hospitalized COVID-19 patients and in 30 healthy volunteers. Compared to the healthy controls, the optical density (OD) level of autoAbs in COVID-19 showed considerable variability distributing over a range between 0.13 and 0.85. We thus divided the patients into two groups, one with OD &gt;0,29 and the other one with a OD &gt;0,29, where 0,29 represents the OD mean value of autoAbs against BPIFB4 in physiological conditions.

Results
Since patients with higher OD are mainly those who spend in average a higher number of days in hospital, we stratified the patients according to the Length of Stay (LoS) in hospital (Figure 1), and found a trend towards a positive correlation between AutoAbs OD level and length of hospitalization within COVID-19 patients.
When present, autoAbs exclusively target the WT-BPIFB4 autoantigens and neglect the recognition of the Longevity-associated-variant-(LAV) of the BPIFB4 gene known for its therapeutic efficacy in cardiomyopathy, atherosclerosis (4), diabetes (6) and platelets' reactivity.
As expected, the pre-treatment of human PrP with the recombinant rhLAV-BPIFB4 reduces platelets' aggregation in response to ADP and collagen in COVID-19 patients in vitro.
On the other hand, at functional level, the well established LAV-BPIFB4-regulated M2 macrophage polarization (4,7), is neutralized in presence of anti-BPIFB4 autoAbs-enriched plasma.

Conclusion
We conclude that a significant proportion of hospitalized COVID-19 patients displays BPIFB4-AutoAbs which are positively correlated with the Length of Stay (LoS) in hospital. In future, it will be of utmost importance to clarify if the 4 missense SNPs which distinguish LAV-BPIFB4 gene from its WT-counterpart, are instrumental to prevent the self-tolerance brake-down and the potential development of specific antibodies against endogenous cardiovascular protectors.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Cariplo Foundation (n.2016-0874) to AAP and CV; Ministry of Health (RF-2016-02364864) to AAP and CV
Hepatic T1-time predicts cardiovascular risk in all-comers referred for cardiovascular magnetic resonance
Authors: Mascherbauer K , Dona C , Koschutnik M , Dannenberg V , Nitsche C , Duca F , Beitzke D , Loewe C , Waldmann E , Trauner M , Bartko P , Goliasch G , Mascherbauer J , Hengstenberg C , Kammerlander A 
Published: October 2022

Abstract

Background
Liver damage is frequently observed in patients with cardiovascular disease (CVD) but infrequently quantified. We hypothesized that in patients with CVD undergoing cardiac magnetic resonance (CMR), liver T1-times indicate liver damage and are associated with cardiovascular outcome.

Methods
We measured hepatic T1-times, displayed on standard cardiac T1-maps, in an all-comer CMR-cohort. At the time of CMR, we assessed validated general liver fibrosis scores. Kaplan-Meier estimates and Cox-regression models were used to investigate the association between hepatic T1-times and a composite endpoint of non-fatal myocardial infarction, heart failure hospitalization, and death.

Results
1022 participants (58±18 y/o, 47% female) were included (972 patients, 50 controls). Hepatic T1-times were 590±89ms in patients and 574±45ms in controls (p=0.052). They were significantly correlated with cardiac size and function, presence of atrial fibrillation, NT-pro-BNP levels, and gamma-glutamyl-transferase levels (p&lt;0.001 for all). During follow-up (58±31 months), a total of 280 (29%) events occurred. On Cox-regression, high hepatic T1-times yielded a significantly higher risk for events (adj.HR 1.66 [95% CI: 1.45–1.89] per 100ms increase, p&lt;0.001), even when adjusted for age, sex, left and right ventricular ejection fraction, NT-proBNP, and myocardial T1-time. On restricted cubic splines, we found that a hepatic T1-time exceeding 610ms was associated with excessive risk.

Conclusion
Hepatic T1-times on standard CMR scans were significantly associated with cardiac size and function, comorbidities, natriuretic peptides, and independently predicted cardiovascular mortality and morbidity. A hepatic T1-time &gt;610ms seems to indicate excessive risk.

Funding Acknowledgement
Type of funding sources: None.
LncRNA BACE1-AS: a link between heart failure and Alzheimer's disease
Authors: Greco S , Piella S N , Made' A , Tascini A S , Garcia-Manteiga J M , Castelvecchio S , Menicanti L , Martelli F 
Published: October 2022

Abstract

Background
BACE1-antisense RNA (BACE1-AS) is a lncRNA antisense to the Beta-Secretase-1 (BACE1) gene encoding a key enzyme in the production of the β-amyloid peptide, associated to Alzheimer's disease (AD). In a previous study1, we showed that the BACE1-AS/BACE1 axis is activated in heart failure (HF) patients, leading to β-amyloid accumulation in failing hearts. Accordingly, BACE1-AS expression in different cultured cardiac cell types induced the expression of BACE1 and β-amyloid that, in turn, triggered apoptosis. The mechanisms underlying BACE1-AS action are not yet fully elucidated. Indeed, current models based on miRNA “sponging” or “masking” inducing BACE1 post-transcriptional stabilization may not recapitulate all BACE1-AS functions.

Purpose
Our aim is to investigate the molecular mechanisms regulated by BACE1-AS in heart failure disease.

Methods
BACE1-AS-pull-down, followed by RNA-Seq, was used to identify RNAs interacting with BACE1-AS in AC16 cardiomyocytes. Moreover, accessible chromatin sites induced by BACE1-AS overexpression were assayed by ATAC-Seq.

Results
BACE1-AS pull-down identified 698 BACE1-AS interacting RNAs. Among these enriched transcripts, 69 were mapping to genomic enhancer regions that have been found to be hypo-methylated in AD brains2. After qPCR validation of the interactions identified by RNA-seq, the modulation of SEMA4D, ABCG1, GFRA2 and RIMBP2, which were under the control of a subset of these enhancers, was assayed upon BACE1-AS overexpression in cardiomyocytes. It was found that their expression was induced, supporting the functional interaction between BACE1-AS and the identified enhancer loci.
To gain further insight into BACE1-AS function in cardiomyocytes, accessible chromatin sites induced by BACE1-AS overexpression were assayed by ATAC-Seq. Interestingly, 17 regions identified by this approach overlapped with enhancers that are hypo-methylated in AD brains. One of the accessible chromatin sites was the locus encompassing RNF214/BACE1/BACE1-AS that maps to active enhancer marks, indicating that BACE1-AS may regulate the expression of BACE1 transcriptionally.

Conclusion
Collectively, these data suggest BACE1-AS as a node of shared disease-mechanisms between heart failure and AD.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Italian Ministry of Health
Enhancing NO pathway improves diastolic function in a hamster model of heart failure with preserved ejection fraction associated with nonalcoholic steatohepatitis
Authors: Dubroca C , Maupoint J , Costard C , Brousseau E , Briand F , Sulpice T 
Published: October 2022

Abstract

Background and aims
Heart failure with preserved ejection fraction (HFpEF) is a complex clinical phenotype frequently associated to the presence of multiple non-cardiac comorbidities including nonalcoholic steatohepatitis (NASH) commonly seen in metabolic syndrome and obesity. In this context, the enhanced production of ROS due to systemic inflammatory state participates to the myocardial structural and functional alterations by limiting nitric oxide bioavailability. We thus evaluated the effect of the phosphodiesterase type 5 (PDE5) inhibitor, vardenafil, on cardiac function and remodeling in a NASH hamster model.

Method
Male Golden Syrian hamsters were fed for 20 weeks with a free choice diet consisting in free access to control chow diet with normal drinking water or a high fat/high cholesterol diet with 10% fructose enriched drinking water. At 15 weeks hamsters were randomized on E/A ratio and ejection fraction to receive either vardenafil (10mg/kg/day orally for 5 weeks) or vehicle before cardiac and metabolic parameters evaluation.

Results
After 20 weeks of free choice diet, hamsters presented a large increase in hepatic lipid contents and developed NASH. Echocardiography showed that systolic function was preserved with ejection fraction in normal range whatever the diet. In contrast, E/A ratio was significantly increased in free choice fed hamsters (2,1±0,1 vs 1,4±0,04) and E'/A' ratio inverted when compared to control (0,8±0,02 vs 1,3±0,04). This was associated with a decreased isovolumic relaxation time (IVRT) and an increased E/E' ratio signing relaxation impairment and higher filling pressure, respectively. Moreover, pro-BNP was largely increased in free choice fed hamsters when compared to controls. While vardenafil had no impact on NASH, diastolic function was largely improved as evidenced by the significant decrease in E/A and E/E' ratios along with the increase in E'/A' ratio and IVRT. This effect was corroborated by the normalization of the pro-BNP plasma levels induced by the 5 week-treatment with vardenafil.

Conclusion
The present data demonstrate that free choice fed hamsters developed NASH and showed the ultimate stage of diastolic dysfunction characterized by a restrictive profile combining compliance and relaxation impairments as well as higher filling pressure. While no effect on NASH was observed, enhancing NO pathway with vardenafil treatment significantly improves diastolic function in this hamster model of HFpEF.

Funding Acknowledgement
Type of funding sources: None.
Comparative effectiveness of the new calpain inhibitor NPO-2270 versus enalapril in pressure overload-induced heart failure
Authors: Delgado S , Aluja D , Llenas J , Vazquez E , Barrabes J A , Ferreira I , Inserte J 
Published: October 2022

Abstract

Background
Studies performed in multiple preclinical models support the contribution of the Ca2+-dependent cysteine proteases calpains to ventricular remodelling and heart failure (HF). However, pharmacological calpain inhibition has not yet been tested in patients with HF mainly due to the limitations of available inhibitors.

Purpose
To determine the effect of NPO-2270 (NPO), a new ketoamide derivative calpain inhibitor, in a mouse model of pressure-overload and compare its effectiveness with that of enalapril.

Methods
C57BL6 mice were subjected to transverse aortic constriction (TAC) for 4 weeks. Mice were randomised to receive orally administered NPO or enalapril at the dose of 10 mg/kg/day once a day, or vehicle, starting at day 7 after TAC surgery. The combination of both drugs and the effect of NPO starting 1 day after surgery were tested in additional TAC mice. Echocardiographic data, markers of hypertrophy, fibrosis, calpain activity and cleavage of calpain substrates were measured at different time points.

Results
TAC increased calpain-1 and -2 expression and activity. Administration of NPO and enalapril prevented the progression of hypertrophy and interstitial fibrosis induced by TAC with no statistically significant differences between the two treatments. However, ongoing ventricular dysfunction was less severe in the NPO group than in the enalapril group (27% of LVEF reduction in control group, 6% in NPO group and 16% in the enalapril group after TAC, P=0.024 between NPO and enalapril groups). The combined treatment or the administration of NPO from the first day after TAC surgery was not superior to NPO alone starting 7 days after TAC. These differences in LVEF correlated with better preservation of cadherin-based cell adhesion complex in mice treated with NPO-2270. No adverse effects associated with long-term NPO administration were observed in a sham group.

Conclusions
The new calpain inhibitor NPO-2270 prevents the development of hypertrophy and fibrosis with similar efficacy than enalapril but prevents cardiac dysfunction more effectively in a preclinical model of pressure overload when given orally at equivalent doses.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Instituto de Salud Carlos III of the Spanish Ministry of Health
Transcriptomic and proteomic profiling of human diabetic heart disease
Authors: Gollmer J , Potter L , Vosko I , Tomin T , Birner-Gruenberger R , Von Lewinski D , Sedej S , Scherr D , Wende A R , Rainer P , Zirlik A , Bugger H 
Published: October 2022

Abstract

Studies in animal models demonstrated the capability of type 2 diabetes (T2D) to induce cardiac dysfunction in the absence of vascular disease. However, whether and how T2D also impairs structure and function in human hearts remains poorly understood. Here, we performed transcriptional and proteomic profiling of left ventricular samples of 8 subjects with T2D, preserved EF (63,5%) and no history of ischemic heart disease (= diabetic cardiomyopathy; DbCM), 7 subjects with T2D, reduced EF (26,9%) and ischemic heart disease (= diabetic heart failure; DbHF), and 15 non-diabetic individuals with normal EF (64,7%) serving as controls. Among 1168 proteins identified by LC-MS/MS, 146 proteins were differentially regulated in DbHF, but only 66 in DbCM. Pathway analysis revealed downregulation of energy metabolic proteins, but upregulation of proteins involved in oxidative stress and inflammatory response. In DbCM, pathways of structural remodeling, cardiomyocyte proliferation, and mechanotransduction were upregulated. Bulk RNA sequencing revealed 1795 differentially regulated genes in DbHF, and 527 in DbCM, with only 128 genes being commonly regulated. DbHF, but not DbCM, could be clearly discriminated from controls by hierarchical clustering. While inflammation/immunity were major regulated pathways in DbHF, extracellular matrix remodeling and cellular growth were the most regulated pathways in DbCM. Thus, the differential regulation of biological pathways in DbCM versus DbHF suggests the existence of two distinct disease entities rather than DbHF being an advanced disease stage of DbCM.

Funding Acknowledgement
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Austrian Diabetes Society
A transgenic mouse model mimicking human cardiac amyloidosis
Authors: Fauvel C , Mulder P , Heron C , Nicol L , Brakenhielm E , Bellien J , Kalopissis A , Bauer F 
Published: October 2022

Abstract

Background
Heart failure (HF) pathophysiology complicating cardiac amyloidosis (CA) is poorly explored due to the lack of relevant animal model. A recently transgenic mouse expressing an amyloidogenic variant of human apolipoprotein AII was developed that showed ubiquitous amyloid deposit but limited data on cardiac involvement.

Purpose
To investigate heart structure and function in transgenic mice expressing an amyloidogenic variant of human apolipoprotein AII

Methods
Seventy-nine mice ageing 2 to 3 months were included in this study as follow: amyloidosis group (n=44) and sham (i.e. genetic mutation without phenotypic expression, n=35). Both were serially imaged by echocardiography (Vevo3100 Fujifilm) and cardiac magnetic resonance imaging (CMR, Biospec 4.7 tesla), and invasively explored by left–sided catheterization (pressure-volume loop [PV loop], Millar Catheterization), before sacrifice and histological investigations.

Results
As soon as 2–3 months of age, the amyloidosis group demonstrated significant left ventricular (LV) hypertrophy, diastolic dysfunction and left atrial dilatation compared with sham group (p&lt;0.01). Left ventricular ejection fraction was initially normal in both groups but deteriorated in amyloidosis mice (p&lt;0.001) before right ventricular function collapses (p&lt;0.001). In amyloidosis group, PV loops showed significant LV end-diastolic pressure increase (p&lt;0.001), stiffer LV (p&lt;0.01) and reduced systolic function LV elastance (p&lt;0.05). Invasive and non-invasive abnormalities paralleled severe CA deposits and subendocardial fibrosis matrix remodeling, both labeling by Congo red and Red Sirius coloration (p&lt;0.001). CMR analysis showed significant T1 (p&lt;0.05) and T2 (p&lt;0.01) signal increase and significant decrease in myocardial perfusion (p&lt;0.01) in the amyloidosis group. Clinically, mice with amyloidosis covered less distance during exercise test (p&lt;0.001) and died earlier (log-rank test, p&lt;0.01).

Conclusion
By mimicking human cardiac amyloidosis, the model of transgenic mouse expressing an amyloidogenic variant of human apolipoprotein AII is promising to investigate the underlying pathophysiology of heart failure due to amyloidosis.

Funding Acknowledgement
Type of funding sources: Private grant(s) and/or Sponsorship. Main funding source(s): Pfizer grant