tbuckley/cabot-search huggingface dataset:
Novel method for induction of myocardial infarction in rats by ultrasound guided electrocoagulation of the left anterior descending artery
Authors: M Nordfalk, S S Groenager, M Flethoej, I Hunter, R S Ripa, A Kjaer, P J Pedersen
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Private company. Main funding source(s): Minerva Imaging Background Murine models of acute myocardial infarction (MI) are commonly established through ligation of the left anterior descending artery (LAD). Despite inherent procedural challenges, including high mortality rates, limited improvements of the induction procedure have been made. Ultrasound-guided electrocoagulation of the LAD could be advantageous by improving animal welfare and decreasing procedural mortality. Purpose To investigate the feasibility of MI induction by ultrasound guided electrocoagulation in rats in comparison to traditional LAD ligation. Methods Thirty-eight female Sprague Dawley rats were randomized to MI by either of the two induction methods leading to 19 rats in each group. Infarct size and left ventricular ejection fraction (LVEF) were evaluated using positron emission tomography (PET) with [18F]fluorodeoxyglucose ([18F]FDG) 9 days post-MI. Myocardial infarct size was categorized as non-infarcted, mild, moderate, or severe based on [18F]FDG uptake. Animal welfare was assessed twice daily for the initial 48h post-surgery using a behavioural scoring system. Results There was no significant difference in the mortality rate of rats with MI induced by electrocoagulation compared to LAD ligation (16% vs 37%, P=0.27). Among the surviving animals both methods produced an equal number of myocardial infarcts (n=5). Electrocoagulation resulted in 1 mild, 2 moderate, and 2 severe MIs, while LAD ligation produced 1 mild, 0 moderate, and 4 severe MIs. Infarct size was similar in the electrocoagulation group compared to the LAD ligation group (mean±SD electrocoagulation: 15±5% vs ligation: 28±13%, P=0.13). The number of non-infarcted surviving rats in the electrocoagulation group was 11 compared to 7 in the LAD ligation group (P=0.33). The LVEF of infarcted rats showed no significant difference between groups (electrocoagulation: 47±8% vs ligation: 38±10%, P=0.14). The LVEF for non-infarcted rats was 53±5%. Post-surgical animal welfare appeared to be improved in the electrocoagulation group, however, the accumulated behavioural score did not reveal a significant difference (median [interquartile range] electrocoagulation: 0 [0-1] vs ligation 2 [0-2], P=0.19). Conclusion Induction of myocardial infarction by ultrasound-guided electrocoagulation is feasible in rats and should be considered as an alternative to LAD ligation. Further development is needed to improve the rate of success and consistency in infarct size.
The effects of siRNA-mediated knockdown of AP-1 on pulmonary arterial and right ventricular dysfunction associated with cardiac and pulmonary fibrosis in cardiopulmonary disease
Authors: I K Comarita, I L Anghelache, G Tanko, A Georgescu
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Romania’s National Recovery and Resilience Plan, PNRR-III-C9-2022-I8, CF 93/15.11.2022, Financing Contract no. 760063/23.05.2023, and by the Romanian Academy Background Pulmonary hypertension (PH) is a complex multifactorial vascular pathology characterized by an increased pulmonary arterial pressure, vasoconstriction, remodelling of the pulmonary vasculature, thrombosis in situ and inflammation associated with right-side heart failure. Purpose Herein, we explored the potential beneficial effects of treatment with siRNA AP-1 on pulmonary arterial and right ventricular dysfunction and perivascular and interstitial fibrosis in pulmonary artery (PA), right ventricle (RV) and lung in an experimental animal model of monocrotaline (MCT)-induced pulmonary arterial hypertension. Methods Golden Syrian hamsters were divided into: (1) C group; (2) MCT group obtained by a single subcutaneous injection of 60 mg/kg MCT at the beginning of the experiment; (3) MCT- siRNA AP-1 group received a one-time subcutaneous dose of MCT and subcutaneous injections containing 100nM siRNA AP-1, every two weeks. All animal groups received water and standard chow ad libitum for 3 months. Results In comparison with the MCT group, siRNA AP-1 treatment had significant beneficial effects contributing to: (1) a reduction in TGF-β1/ET-1/IL-1β plasma concentrations; (2) a reduced level of cytosolic ROS production in PA, RV and lung, and ultrastructural improvements of these tissues; (3) a decrease of inflammatory and fibrotic marker expressions in PA (COL1A, Fibronectin, Vimentin, α-SMA, CTGF, Calponin, MMP-9), RV (COL1A, CTGF, Fibronectin, α-SMA, F-actin, OB-cadherin) and lung (COL1A, CTGF, Fibronectin, α-SMA, OB-cadherin), and an increase of endothelial marker expressions (CD31/VE-cadherin) in PA; (4) structural and functional recoveries of PA (reduced VTI/Vel, restored vascular response) and RV (enlarged internal cavity diameter in diastole, increased TAPSE/PRVOFs) associated with a decrease in systolic/diastolic blood pressure, and heart rate; (5) a reduced protein expression profile of AP-1S3/ pFAK/FAK/pERK/ERK and a significant decrease in the expression levels of miRNA-145, miRNA-210, miRNA-21, and miRNA-214 along with an increase of miRNA-124 and miRNA-204. Conclusion The siRNA AP-1-based therapy led to an improvement of pulmonary arterial and right ventricular function accompanied by a regression of perivascular and interstitial fibrosis in PA, RV and lung and a down-regulation of key inflammatory and fibrotic markers in MCT-treated hamsters.
Antisense oligonucleotides as a potential therapeutic strategy in hypertrophic cardiomyopathy caused by MYBPC3 variants
Authors: S Munteanu, E M Nicolas Rocamora, C Martinez Perez, A Bernabe Garcia, F J Nicolas Villaescusa, J R Gimeno Blanes, V Arechavala Gomeza, M Sabater Molina
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Sociedad Española de Cardiología Background/Introduction Non-sense or frameshift mutations in the MYBPC3 gene encoding cardiac myosin-binding protein C (cMyBP-C) are a major cause for inherited forms of Hypertrophic Cardiomyopathy (HCM), the most frequent inherited cardiac disease, affecting 0.2% of the general population and represents the leading cause for sudden cardiac death in young athletes. During the last decade, RNA-based therapeutic approaches have been studied for neuromuscular disorders and recently became to be considered to envision causal therapies for cardiomyopathies, utilizing antisense oligonucleotide mediated exon skipping, which attempts to reframe mutated mybpc3 transcripts, resulting in shortened but functional protein. MYBPC3 gene has a total of 34 coding exons and, exons 2, 22, and 24-27 are ideal targets for skipping. Purpose We aim to design and evaluate antisense oligonucleotides (AONs) for efficient exon-skipping in targeted MYBPC3 exons. Methods We designed two specific AONs to mask the MYBPC3 exon 26 and confirmed successfully mediated exon skipping without disrupting the MYBPC3 reading frame. We chose a previously identified MYBPC3 c.2671insG/p.R891Afs*160 variant in exon 26, which causes a frameshift mutation leading to truncated MYBPC3 in HCM. Primary Human Cardiac Myocytes (PHCM) (Promocell) were transfected with the splice-site targeting AONs. Results In this study we designed two chemically modified AONs (AON1 and AON2) through the addition of a 2′-O-methyl RNA phosphorothioate backbone, targeting exonic splicing enhancers (ESEs) or splice sites (SSs) can induce skipping of MYBPC3 exon 26. We evaluated AON1 (i26-3SS) and AON2 (ESE)-treated Primary Human Cardiac Myocytes (PHCM) show 26-27 exons skipping in comparison to control samples. The identity of the exon 26-27 skipped bands detected by RT-PCR was confirmed by Sanger sequencing and preservation of both nucleotide sequences of MYBPC3 exons 25 and 28 were observed. These results indicate that AONs specifically for mutated MYBPC3 target exons, expanding the framework of future advancements in the therapeutic potential of antisense-mediated exon skipping MYBPC3-based HCM. In addition, skipping exons 26 and 27 we eliminate a total of 22 possible exonic mutations, which represent 7.2% of the total known MYBPC3 exonic mutations. Conclusion(s) These results demonstrate that disruption of the MYBPC3 reading frame due to a truncating HCM mutation can be restored by exon skipping in cardiomyocytes in vitro indicating RNA-based strategies as a potential treatment option for HCM.Exon skipping scheme
Differential sex-dependent role of lncRNAs in cardiomyopathy
Authors: D Franco, F Martinez-Amaro, F Bonet, R Toro, I Perez De Castro, F Rochais, L Bouchard, A Salguero, J L De La Pompa, C Garcia-Padilla
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Ministry of Education and Science Cardiovascular diseases are one of the most prevalent causes of mortality and morbidity worldwide. Structural diseases such as dilated and hypertrophic cardiomyopathy significantly contributed to it. In recent years, several studies have pinpointed the importance of non-coding genome in the pathogenesis of cardiomyopathy. We have previously reported the role of distinct lncRNAs in atrial fibrillation. We now explored the plausible role of these lncRNAs in cardiomyopathy by analyzing their expression in two different mouse mutant models of lamin (LMNA) - LMNAR249W and αMHCCreLMNAFF, a cardiac hypertrophic MyBPC3 mutant model as well as in experimental myocardial infarction mice. Importantly, both LMNA mutant models developed a dilated cardiomyopathy, a condition distinctly observed in males but not in females. Our data demonstrated that Walras, Gm26533, Wallrd, Walaa and Walrad are downregulated in females, but not males. Furthermore, differential deregulation is also observed in MyBPC3 mutant model but not in myocardial infarction. In vitro analyses demonstrated an estrogen-dependent regulation. In addition, expression analyses in distinct cardiovascular cell lines revealed enriched myocardial expression of Walras and Gm26533 while Wallrd and Walaa are preferentially expressed in endocardial cells. These data were further validated by in situ hybridization in embryonic cardiac tissues. Walras overexpression resulted in enhanced expression of two key factors of UPR signaling pathway, i.e. Atf6 and Ire1, while Walras inhibition downregulated them. Upregulation of both UPR factors in Walrasgain-of-function assays is accompanied by increased cardiomyocyte apoptosis. In contrast, Walras inhibition resulted in increased Bcl2 expression and therefore increased cell viability. Mechanistically, Walras directly interacts with calumenin protein, a known protective factor against UPR-associated apoptosis, as demonstrated by pulldown assays. In sum, we demonstrated sex-dependent regulation of distinct lncRNAs in different cardiomyopathic mouse models and at least one of them, i.e. Walras contributes to UPR-associated apoptosis regulation, providing a causative link to the development of distinct cardiomyopathies.
Lenti-viral overexpression of RhoGEF Trio reduces CD8 T cell extravasation into atherosclerotic vein graft lesions in mice
Authors: T Sluiter, L Starink, A D Van Stalborch, C Bax, R Schoon, S Huveneers, P H A Quax, A A F De Vries, J D Van Buul, M R De Vries
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Rembrandt Institute for Cardiovascular Sciences Background In advanced atherosclerotic plaques, such as vein graft lesions, neovessels grow into the hypoxic plaque. This intraplaque angiogenic neovessels lack proper endothelial cell-cell junctions and are therefore susceptible to extravasation of leukocytes, which results in inflammation and atherosclerosis. T cells are the largest leukocyte subset found in atherosclerotic plaques and are key drivers of inflammation and plaque progression. RhoGEF Trio improves endothelial barrier function by strengthening endothelial cell-cell junctions in vitro, but its effect on T cell transmigration and its therapeutic potential to impede atherogenesis remains unknown. Methods ApoE3*Leiden mice (n=10/group) underwent bypass surgery in which a donor caval vein was interpositioned into the arterial circulation at the site of the right common carotid artery. Immediately post-operatively, a pluronic gel containing either GFP-Lentivirus (LV) (control) or Trio-GFP-LV (Trio) was locally applied around the vein graft to induce local overexpression of Trio. At post-op day 28, vein grafts were harvested and processed for morphometric and compositional analysis of the atherosclerotic lesions. Additionally, CD4 and CD8 T cells were isolated from healthy human donors and used in vitro to assess the role of Trio on trans-endothelial migration over in vitro-cultured HUVECs. Results At post-op day 28, both luminal and neovessel endothelial cells were GFP positive indicating successful transfection. CD8 T cell infiltration into the atherosclerotic plaque was reduced in the Trio group (45%, p=0.0426), whilst plaque size and the number of neovessels were unaffected. Mechanistically, we observed that adherence of CD8, but not CD4 T cells to the endothelial monolayer was reduced in vitro upon transfection with Trio-LV. This resulted in a decrease in the absolute number of transmigrated CD8 T cells (56%, p=0.0197), whilst the number of transmigrated CD4 T cells was unaffected. Interestingly, CD8 T cells exhibited no difference trans-/paracellular diapedesis, whilst for CD4 T cells Trio overexpression induced transcellular, rather than paracellular, transmigration. Conclusions LV transfection of vein grafts by local application of a pluronic gel holds promise a therapeutic strategy to improve bypass surgery outcomes. LV transfection with RhoGEF Trio did reduce CD8 T cell transmigration (in vivo and in vitro), but not CD4 T cell transmigration (in vitro) into the atherosclerotic plaque, whilst plaque size was not affected. Ongoing investigations focus on the effect of Trio on micro-vascular leakage as well as infiltration of other immune cells into the plaque.
The endocannabinoid ARA-S acts as an endogenous rescuing factor of Kv7.1/KCNE1 carrying Long QT syndrome mutations and paves the way for drug development strategies
Authors: I Hiniesto Inigo, A Sridhar, S A Lundholm, S I Liin
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Union’s Horizon 2020 research and innovation program Long QT Syndrome (LQTS) is a cardiac arrhythmia characterized by delayed ventricular repolarization. LQTS occurs when there are loss-of-function mutations in genes encoding for voltage-gated channels and their beta subunits that contribute to the cardiac action potential. Among them, the most mutated channel is Kv7.1/KCNE1, which encodes for the slowly activating component of the delayed rectifier K+ current (IKs). Families with inherited mutations in the Kv7.1/KCNE1 exhibit inconsistent correlations between genotypes and phenotypes making the clinical management of LQTS challenging. A possible strategy to help guide clinical approaches involves exploring endogenous compounds capable of modulating the Kv7.1/KCNE1 channel function, acting as potential disease modifiers. Using the two-electrode voltage clamp technique, we previously found that the endogenous endocannabinoid-like compound ARA-S, which has an arachidonic acid tail and a serine head group, facilitates activation of the wild-type Kv7.1/KCNE1 channel expressed in Xenopus oocytes. This is seen as a left shift in the voltage dependence of channel opening and an increase in conductance. Moreover, we showed that ARA-S restores a physiological QT interval in isolated guinea pig hearts suffering from drug-induced LQTS. Hence, ARA-S is an interesting endogenous Kv7.1/KCNE1 channel modulator that may influence disease severity. However, it is uncertain whether ARA-S also activates the Kv7.1/KCNE1 when it carries LQTS-associated mutations. Therefore, the current study involves studying the effect of ARA-S on mutated Kv7.1/KCNE1 channels using two-electrode voltage clamp electrophysiology. We find that ARA-S facilitates the activation of all tested LQTS mutants. However, the magnitude of the effect varies among them, suggesting that the compound´s effect is dependent on the genetic background. To understand why ARA-S has different effects, we use computational simulation approaches to study how mutations in the putative ARA-S binding sites change ARA-S efficacy. This understanding aids in developing new substances that can activate different mutations, by, for example, modifying the ARA-S tail to enhance its efficacy. We also try to understand the concentration of ARA-S required to restore a wild-type-like function of each mutant, considering the mutant's intrinsic behavior and the apparent affinity of ARA-S for the specific mutant. Altogether, this work finds ARA-S to be an efficient activator of LQTS-associated mutant Kv7.1/KCNE1 channels and is a first step towards understanding how the genetic background determines the effect of this endogenous modulator.
Transcriptomic insights into cardiac healing - unveiling promising targets for effective gene therapies in diseased hearts
Authors: R Caliandro, J Hanemaaijer, S Van Den Berg, Q Gunst, R J Oostra, M Van Den Hoff, M M Gladka
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Dutch Heart Foundation Identifying therapeutical target genes is the first step towards developing new gene therapies to promote cardiac repair after injury. In the last decades, despite the many steps forward, not a single therapy has shown to be effective and suitable for human treatment of the injured heart. Considering that cardiac diseases represent one of the most prevalent causes of morbidity, disability, and mortality in the world, it is, therefore, essential to develop new and improved therapies to induce cardiac regeneration. In this study, we aimed to identify novel therapeutical targets that could be used for future purposes of cardiac gene therapy. Unlike previous studies that investigated fetal hearts that are physiologically and functionally different from mature hearts, we focused on investigating adult human hearts that we collected from human donors with post-mortem time below 5 hours. Samples of left ventricles were molecularly and morphologically characterized for the presence of stress, cardiac damage, and cardiac fibrosis. Results of the molecular analyses were cross-referenced with the donors' medical history to classify each specimen as "control" or "diseased." Next, we performed single nuclei RNA sequencing of samples of the left ventricle from 2 "control" and 3 "diseased" hearts. Due to the adult tissue's complexity and the specimen's high-fiber content, we optimized the nuclei isolation protocol. Eventually, we successfully sequenced an average of 8.000 nuclei per sample with a gene coverage of approximately 800 genes per nucleus, identifying, via unsupervised cluster analysis, 17 different cell populations. In cardiomyocytes, we identified over 500 genes differentially expressed in diseased hearts when compared to healthy controls. Among these genes, we discovered several novel target genes whose expression is increased only in diseased cardiomyocytes. Gene ontology analysis of the known interactors of the newly identified target genes shows enrichment in terms such as sarcomere organization, muscle contraction, mitochondrial energetics, calcium handling, and fatty acid metabolism. Currently, we are investigating the molecular effects of the gain and loss of function of these genes in healthy iPS-derived cardiomyocytes and human-derived myocardial tissue slices to better understand their role in the pathophysiology of the adult human heart. From a therapeutic perspective, our ongoing research holds promises for developing new and improved intervention therapies to promote cardiac regeneration.
MicroRNA-127-3p regulates inflammatory response in injury-induced vascular remodeling and cellular senescence in vascular smooth muscle cells through mTor
Authors: F Daniel, K Kalies, L Hehl, S Guerlach, J M Koester, S Boettcher, K Knoepp, J Dutzmann, D Sedding
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Martin-Luther-Universität Halle-Wittenberg Background and Purpose The accumulation of senescent cells in the vasculature contributes to impaired vascular function over the lifetime. Micro RNAs (miRs) are promising therapeutic targets due to their cell-specific functions. Targeting miR-127-3p emerges as an attractive approach to enhance vascular healing and limit angioplasty-related complications. Here, we focus on elucidating the role of miR-127-3p in vascular smooth muscle cells (VSMC) to characterize their role in cellular function and vascular senescence. Methods Utilizing reverse transcription and qRT-PCR we checked expression levels of miR-127-3p in human and murine aortic VSMCs. After overexpression and downregulation via transfection of pre-miR and antagomiR, we evaluated the influence of miR-127-3p on cellular functions such as proliferation and migration of human (non-) replicative senescent VSMCs. Potential targets were selected via target prediction analysis and verified on mRNA and protein levels. Morphological changes were visualized via high-resolution fluorescence microscopy. Senescence marker expression was checked after transfection by qRT-PCR. Results MiR-127-3p is highly upregulated in femoral artery tissue on day 10 after wire-induced injury in C57Bl6 mice, serving as restenosis model (p&amp;lt;0.001) and aged C57BL/6 mice (3 months vs. 22 months, p&amp;lt;0.0001). Replicative senescent human VSMC exhibit increased miR-127-3p expression (p&amp;lt;0.05). In vitro, miR-127-3p overexpression reduces proliferation in senescent (p&amp;lt;0.001) and non-senescent (p&amp;lt;0.01) cells. It hinders migration in non-senescent cells (p&amp;lt;0.0001) but has a reverse effect in senescent cells (p&amp;lt;0.01). Potential targets affecting cellular functions, inflammatory pathways, and cellular aging were identified as mTOR, AXL, and TEAD3 (p&amp;lt;0.05). Transfection of pre-miR leads to increase in cell size and the acquisition of a flattened morphology of human VSMC. P14arf and laminB1, as senescence-associated genes, showed downregulation after overexpression of miR-127-3p on mRNA level (p&amp;lt;0.05). When VSMCs were stimulated with anti-miR-127-3p, increased mRNA levels of p14arf and laminB1 were detected (p&amp;lt;0.05). Conclusion This study identifies miR-127-3p as a contributor to cellular functions including cellular proliferation and migration and seems to have opposing effects in non- and replicative senescent VSMC. Further research is needed to validate miR-127-3p as a therapeutic target for vascular aging and remodeling in vitro and in vivo.