bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of RhoA and Cdc42 by miR-133a modulates retinoic acid signaling during early development of the posterior cardiac tube segment
Authors: Lopez-Sanchez C , Garcia-Padilla C , Garcia-Lopez V , Aranega A , Franco D , Garcia-Martinez V 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public Institution(s). Main funding source(s): Junta de Extremadura with FEDER co-financing.

Introduction
It has been demonstrated that multiple microRNAs play crucial roles in diverse biological processes, including cardiovascular development. In particular, we have previously observed miR-133a expressing at different stages of the primitive streak precardiac cells until the formation of the cardiac tube. MiR-133a role has also been reported as a modulator of proliferation and differentiation during heart development, involving many target genes and signaling pathways. Furthermore, we have shown in previous studies that retinoic acid plays a crucial role in cardiac development, modulating the expression of atrial markers.

Purpose
The aim of this work is to analyze the miR-133a modulation role on the retinoic acid signaling pathway in the posterior segment of the heart tube during cardiac looping formation.

Methods
By means of microinjections in both primitive endocardial tubes of chick embryo culture, gain- and loss-of-function experiments were performed with premiR-133a and anti-miR-133a, respectively. Subsequently, embryos were subjected to whole mount in situ hybridization with Tbx5 and AMHC1 probes, and qPCR. After TargetScan analysis and luciferase assays, immunohistochemistry was carried out by using anti-RhoA, -Cdc42 and -Raldh2/ALDH1A2 antibodies.

Results
Luciferase assays show that miR-133a is capable to recognize and bind to RhoA and Cdc42 3´UTR, thus triggering their mRNA degradation. Our results reveal that miR-133a gain-of-function experiments repress the expression of atrial markers Tbx5 and AMHC1, as well as RhoA, Cdc42 and -Raldh2/ALDH1A2 signals. Supporting these data, miR-133a loss-of-function experiments show the opposite effect: they increase the expression of Tbx5 and AMHC1, as well as RhoA, Cdc42 and Raldh2/ALDH1A2 signals.

Conclusion
Our results suggest that miR-133a could modulate negatively retinoic acid signaling by triggering degradation of pivotal genes RhoA and Cdc42, both required for Raldh2/ALDH1A2 expression, crucial to the synthesis of retinoic acid.
The euchromatic histone-lysine N-methyltransferases EHMT1/2 suppress post-natal cardiomyocyte proliferation via histone 3 lysine 9 (H3K9) dimethylation
Authors: Chatzieleftheriadis K , Sadredini MS , Donate Puertas RDP , Sjaastad IS , Roderick LR 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – EU funding. Main funding source(s): KULeuven C14/17/088
Flemish Research Foundation (FWO) G0C6419N
Cardiomyocytes proliferate rapidly during developmental growth in utero, but this proliferative capacity dramatically declines postnatally, rendering the heart unable to repair itself subsequent to injury or during ageing. The mechanisms governing this postnatal loss of cell cycle activity are not fully understood. During maturation, epigenomic remodelling drives wholesale changes in the cardiomyocyte transcriptome from the neonatal to the adult state. This is associated with silencing of foetally-expressed genes and activation of expression of genes required for adulthood. Here, we tested the hypothesis that epigenomic remodelling silences cell cycle-associated genes and hence cell cycle activity postnatally. In this regard, we focused on the role of the repressive H3K9me2 mark.
Using immunostaining of Ki-67 and Aurora Kinase B as well as analysis of EdU incorporation to identify cell cycle activity in primary cardiomyocytes culture and in cryosections from rat hearts spanning from embryonic day 18 to adulthood, the abundance of H3K9me2 was found to be inversely correlated with cell proliferation. In line with these data, the expression of a gene panel associated with cell cycle activity decreased postnatally and was inversely related with H3K9me2 abundance in cardiomyocyte nuclei, purified by flow cytometry based on H3K9me2 abundance. Inhibition of the euchromatic histone lysine methyltransferases (Ehmt1/2 aka GLP/G9a), with the highly selective inhibitor A-366, enhanced proliferation of neonatal rat cardiomyocytes in vitro, as well as in vivo, in neonatal mice, via concomitant reduction of the repressive histone mark, H3K9me2. Similar effects were observed in vitro by overexpression of the histone di/tri-methyl demethylase KDM4B. Notably, forced induction of cell cycle activity by overexpression of cyclin/cyclin-dependent kinase significantly reduced H3K9me2 levels, in vitro. Together, these data point to a key role of H3K9me2 and its deposition by Ehmts in post-natal suppression of proliferation. Targeting this repressive mark could provide a strategy to enhance cardiac repair for therapeutic benefit.
Assessment of proarrhythmogenic risk for cannabidiol using dog and rabbit cardiac preparations: the electrophysiological effects on action potential and transmembrane potassium currents
Authors: Naveed M , Topal L , Prorok J , Paszti B , Csupor D , Baczko I , Virag L , Jost N , Varro A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public Institution(s). Main funding source(s): Economic Development and Innovation Operative Programme GINOP-2.3.2-15-2016-00012, the National Research Development and Innovation Office (NKFIH K 135464 and NKFIH K 128851), the Ministry of Human Capacities Hungary (20391-3/2018/FEKUSTRAT and EFOP-3.6.2-16-2017-00006), and from the Eötvös Loránd Research Network

Background
Cannabidiol (CBD), a major active phytogenic cannabinoid, is one of the main constituents of cannabis. Cannabis has been widely used as recreational drug over the decades and its use is constantly increasing as hallucinogenic and/or medicinal agent. However, significant cardiovascular side effects can accompany its use ranging from arrhythmia to sudden cardiac death.

Purpose
The aim of the present work was to investigate the possible cardiac adverse electrophysiological effects of cannabidiol (CBD) on action potentials and various transmembrane potassium currents, such as the rapid (IKr) and slow (IKs) delayed rectifier, the transient outward (Ito) and inward rectifier (IK1) potassium currents in rabbit and dog cardiac preparations to assess the cardiac safety profile and proarrhythmic risk.

Methods
In the current study, conventional microelectrode and voltage clamp techniques were used to record the action potential and transmembrane ionic currents in dog and rabbit ventricular tissue preparations and enzymatically isolated myocytes, respectively.

Results
The results show that CBD lengthens APD90 significantly at the concentration of 5 µM both in dog and rabbit ventricular tissues without changing other action potential parameters significantly. To further investigate the APD90 lengthening effect of CBD, transmembrane potassium currents (IKr, IKs, Ito and IK1) were investigated in dog and/or rabbit ventricular myocytes using voltage clamp technique. CBD significantly inhibited IKr and IKs currents in rabbit ventricular myocytes with an estimated EC50 values of 4.9 and 3.1 µM, respectively. The effect of CBD on rabbit’s Ito current was not significant while it was significant on dog’s Ito current with an estimated EC50 value of 5 µM. IK1 was not responsive to CBD even at high concentration.

Conclusion
In conclusion, looking at the inhibitory effects of CBD on repolarizing potassium currents, despite of the fact that these calculated EC50 values are higher than pharmacokinetics based Cmax values of CBD recorded after smoking and oral intake, it can be speculated that in the presence of certain cardio active drugs or co-morbidity where CBD metabolism or cardiac repolarization reserve is impaired CBD can have an additive and proarrhythmic effect.
Mitochondrial function after endogenous glutathione synthesis stimulation in old rats heart
Authors: Okhai I , Fedichkina R , Goshovska Y , Sagach V 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.
Aging is accompanied by alterations mitochondrial function, and decreasing mitochondrial membrane potential (Δψm) amid increasing ROS/RNS. Mitochondrial disfunction is one of the cause of age-relating cardiovascular deseases development. Previously, we showed that stimulation synthesis of antioxidant glutathione (using DL-propargylglycine (PAG) + L-cysteine) provided complete restoration of the heart and mitochondrial function, reduced ROS levels at ischemia-reperfusion model in adult rats.
The aim was to investigate whether PAG+L-cysteine stimulated glutathione synthesis in old rats, and cardiac Δψm, NOx (nitrite plus nitrate) and ROS production at endogenous stimulation glutathione synthesis.
Stimulation of glutathione synthesis was performed by combined pretreatment with 11.3 mg/kg PAG and 120 mg/kg L-cysteine injected intraperitoneally 40 and 30 min before sacrificing. Langendorff isolated rat hearts were underwent 20 min perfusion. Cardiac mitochondria were isolated and tested for respiration rate and Δψm with TPMP+-selective electrode. Proton-leak was titrated with malonate addition. Glutathione level was measured by Ellman's reagent. Generation rate of superoxide radical and hydrogen peroxide, malondialdehyde and NOx levels, iNOS and cNOS (inducible and constitutive nitric oxide synthase) activity was determined spectrophotometrically in heart tissue.
Pre-administration of PAG+L-cysteine to old rats increased the reduced glutathione level by 51.4%, P&lt;0.05; significantly increased Δψm (P&lt;0.01) and simultaneously decreased the rate of oxygen consumption by mitochondria (P &lt;0.001); the proton-leak curve shifted to the right. It indicates improving the mitochondrial efficiency. At the same time, ROS generation was reduced by 48.2% and 21.7% for superoxide radical and hydrogen peroxide correspondingly, P&lt;0.01. Malondialdehyde level dropped almost twice (P&lt;0.01) too. Quantity NOx falled off by 24.5% with 52.5% decreased activity of iNOS and increased activity cNOS by 2.2 times, P&lt;0.01. The reducing superoxide production and this condition of NO system may indicate of increase the coupling cNOS and ability heart to relax.
Thus, endogenous glutathione synthesis stimulation in conditions of aging caused increasing Δψm and improved respiratory chain efficacy. The redox balance and metabolism of nitric oxide were normalized in rat heart.
over-activated p21 protein activated kinase-3 (pak3) leads to myocardial lipotoxicity and cardiac dysfunction in diet-induced metabolic stress
Authors: Liu W , Ruiz-Velasco A , Raja R , Kaur N 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): British Heart Foundation

Background
In clinics, metabolic stress-induced cardiomyopathy is featured as diastolic/systolic dysfunction, at least partially resulting from impairment of metabolic flexibility in the myocardium, which leads to lipotoxicity and oxidative stress accompanied with cardiomyocytes death. P21-activated kinases (Paks) are a family of serine/threonine kinases, involved in cell survival, proliferation and cytoskeleton remodelling. In spite of the cardioprotective functions of Pak1 and Pak2 in the myocardium, the role of cardiac Pak3 is unexplored.

Purpose
The study is to investigate the function of cardiac Pak3 on myocardial steatosis in response to diet-induced metabolic stress, which will provide proof-of-concept evidence for the therapeutic potential for alleviating cardiac dysfunction by targeting lipotoxicity in the myocardium.

Methods
Pak3 overexpression in the myocardium was achieved by tail vein injection of adeno associated virus (AAV9) delivery of troponin T promoter-driven Pak3 on C57BL/6J mice. The mice were fed with high fat high sucrose diet, the metabolic profiles and cardiac function were measured at 4-week intervals. In addition, cardiac morphological and cellular changes were assessed, including cell size, fibrosis formation, cell death, lipid accumulation and lipid peroxidation. In addition, the transcript levels of the genes either participating lipid metabolism or regulating oxidative stress were also assessed by qPCR.

Results
First, we detected that Pak3 expression and activation was increased in the myocardium from human with obese or diabetes. Consistently, the augmented Pak3 was also detected in the failing heart from C57BL/6J mice, ob/ob and db/db mice. Next, in a time-course study, the mice with AAV9-delivery cardiac Pak3 overexpression were vulnerable to cardiac dysfunction upon diet-induced metabolic stress. Pak3 overexpression did not alter cardiomyocyte size; however, more fibrosis and cell death in the myocardium were detected due to Pak3 enhancement. More importantly, myocardial lipid accumulation and profound lipid peroxidation were determined by immunefluorescent staining of Perilipin 5 (PLIN5) and 4-Hydroxynonenal (4-NHE), respectively. Finally, the abnormal expression of the genes involved in lipid synthesis and fatty acid oxidation was detected in Pak3-overexpressed myocardium under metabolic stress, whereas the levels of anti-oxidative stress genes were decreased.

Conclusion
Altogether, the study has demonstrated that Pak3 over-activation makes the heart more susceptible to myocardial lipotoxicity, which accelerates cardiac steatosis and dysfunction in the fact of metabolic stress.
Temporal and spatial dynamics in the regulation of myocardial metabolism during the ischemia-reperfusion process
Authors: De Dios E , Rios Navarro C , Moya J , Gavara J , Perez-Sole N , Marcos-Garces V , Forteza MJ , Diaz A , Ruiz-Sauri A , Chorro FJ , Bodi V 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): This work was supported by “Instituto de Salud Carlos III” and “Fondos Europeos de Desarrollo Regional FEDER” (grant numbers PI20/00637 and CIBERCV16/11/00486) and Conselleria de Educación – Generalitat Valenciana (PROMETEO/2021/008).

Introduction
In the context of severe myocardial ischemia, cardiac metabolism shifts from beta oxidation to glycolisis. However, the temporal and spatial dynamics of the main regulators of myocardial metabolism during the ischemia-reperfusion process in the infarcted heart has not been fully characterized.

Methods
Myocardial infarction (MI) was induced in swine by means of 90 minutes occlusion of the mid left anterior descending coronary artery using angioplasty balloons. Tetrazolium staining and intracoronary infusion of thioflavin-S were used to define the infarcted, adjacent, and remote areas. mRNA and protein expression of PGC1a, PPARa, ERRa, GLUT1, and GLUT4 were quantified in controls and in MI groups submitted to 48 hours and 3 weeks of reperfusion.

Results
Compared to controls, a severe and generalized drop of PGC1a mRNA gene and protein levels occurred in the infarcted, adjacent and remote areas since ischemia onset until 48 hours reperfusion that persists at 1 month in the infarcted region. Similar dynamics occurred in the infarcted, adjacent, and remote areas in the case of PPARa gene expresion; PPARa protein significantly decreased only until 48 hours reperfusion in the infarcted area. ERRa gene and protein expression persistenly decreased only in the infarcted region since ischemia onset until 1 month. Incrases in GLUT1 (since ischemia onset) and GLUT4 (at 1 month) were detected.

Conclusions
Dynamics and generalized changes in metabolism regulation to a shift from beta oxidation to glycolisis occur in the infarcted heart since ischemia onset until late after reperfusion. Further research in this field can be helpful for a better understanding of pathophysiology of myocardial infarction and to explore new therapeutic options.
Adipsin a regulatory protein found in the extracellular matrix is modulated by hypercholesterolemia in myocardial infarction
Authors: Padro T , Garcia-Arguinzonis M , Vilahur G , Diaz-Riera E , Badimon L 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public Institution(s). Main funding source(s): Agencia Estatal de Investigación (AEI)- Spanish Institute of Health Carlos III, ISCIII;
Agencia Estatal de Investigación (AEI)- Spanish Ministry of Economy Competitiveness of Science

Background
Local inflammation and innate immunity are key features during cardiac remodelling post myocardial infarction (MI). Adipsin, a component of the inflammatory response, is the major activator of the C3-complement pathway and exerts elastolytic activity. Beyond liver, adipsin is mostly secreted from adipocytes, monocytes and macrophages. It is to notice that intramyocardial lipid accumulation alters the fibrotic reparative process of the scar, resulting in larger infarcts and cardiac dysfunction in a pig model. Adipsin could directly contribute to collagen turnover in the cardiac extracellular matrix (ECM) during formation of the post-MI scar.

Purpose
The role of adipsin in the setting of myocardial infarction remains unknown. Here we investigated adipsin changes (content and activity) in the cardiac ECM of MI pigs and determine whether hypercholesterolemia relates to the turnover of the ECM-collagen pattern in association with the adipsin content.

Methods
Normo- (NC) and diet-induced hyper- (HC) cholesterolemic animals (N=28) were randomly distributed to MI-induction or sham-operation (N=7 animals/group). MI was induced by 90min closed-chest coronary balloon occlusion. After revascularization, animals were either maintained for 2.5hours or 30days before being sacrificed. Adipsin, C3- activation products (as readout of adipsin activity) and collagen fragmentation products were determined in cardiac ECM-extracts from the ischemic and non-ischemic regions in all pigs.

Results
Adipsin and C3-activation products (iC3b, C3dg) were consistently detected in the cardiac ECM of sham-operated animals. Detection levels of adipsin, iC3b and C3dg were &gt;2fold higher in HC vs NC pigs (p&lt;0.05) suggesting higher adipsin activity in hearts of animals with high cholesterol levels. NC and HC MI-pigs subjected to 2.5hours reperfusion showed an acute and sharp increase in cardiac adipsin detection in the ischemic region as compared to sham and non-ischemic areas (p&lt;0.05). At 30days post-MI, adipsin content in the scar was comparable to that of sham-operated animals remaining significantly higher in HC vs NC pigs. Yet, adipsin cardiac levels in the scar of HC animals were associated with a lower content of fibrillar collagen-1 (MW: 235kDa) and a higher proportion of proteolytic collagen-fragments (MW: 135-48 kDa) (p&lt;0.05 vs NC pigs)

Conclusions
Adipsin is a component of the cardiac ECM. Hypercholesterolemia enhances myocardial adipsin content and activity and promotes proteolysis of collagen fibrils in the forming scar contributing to the cardiac remodeling during post-infarction.
Sex-related differential susceptibility to ponatinib-induced cardiotoxicity and its relationship to modulation of Notch signaling in a murine model
Authors: Madonna R , Pieragostino DP , Pieragostino DP , Cufaro MC , Cufaro MC , Del Boccio PD , Del Boccio PD , Pucci AP , Pucci AP , Mattii LM , Mattii LM , Doria VD , Doria VD , Cadeddu Dessalvi CD , Cadeddu Dessalvi CD , Zucchi RZ , Zucchi RZ , Mercuro GM , Mercuro GM , De Caterina RD , De Caterina RD 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.

Background
Ponatinib (PON), a tyrosine kinase inhibitor approved in chronic myeloid leukemia, has proven cardiovascular toxicity. Although sex is a risk factor for PON-induced cardiotoxicity in humans, little is known about its mechanisms in general, and sex-related mechanisms in particular.

Objectives
To determine the mechanisms of sex-related PON-induced cardiotoxicity and identify potential rescue strategies in a murine model.

Methods
24-months-old male and female C57B5 mice were treated with 3 mg/kg/day of PON or vehicle via oral gavage for 28 days, with/without siRNA-Notch1 or siRNA-scrambled via tail vein every 3 days.

Results
PON + scrambled siRNA-treated male mice had a higher number of TUNEL-positive cells, a higher percentage of senescence-associated β-galactosidase positive senescent cardiac areas, as well as a lower reactivity degree for the survival marker Bmi1 than female counterparts. Proteomics analysis of cardiac tissue showed upstream activation of nitric oxide synthase (NOS) type 2, downstream activation of cell death and production of reactive oxygen species in PON + scrambled siRNA- compared to vehicle or PON + Notch1 siRNA-treated male mice. Upstream analysis showed beta-estradiol activation, while downstream analysis showed activation of cell survival and inhibition of cell death in PON + scrambled siRNA compared to vehicle-treated female mice. PON + scrambled siRNA-treated mice also showed a downregulation of cardiac actin, which was more marked in male; as well as vessel density, which was more marked in female mice. Female hearts showed a greater extent of cardiac fibrosis than male counterparts at baseline, with no significant changes after PON treatment. In contrast, PON + scrambled siRNA-treated mice had less fibrosis than vehicle or PON + Notch1-siRNA-treated mice. Left ventricular systolic dysfunction shown in PON + scrambled siRNA-treated male mice and - to a lesser extent - by female mice was similarly reversed in both PON + Notch1-siRNA-treated male and female mice (Table 1).

Conclusions
We found a sex-related differential susceptibility and Notch1 modulation in PON-induced cardiotoxicity. This can improve our understanding of sex-related differences and help identify biomarkers in PON cardiotoxicity.