tbuckley/cabot-search huggingface dataset:
Leukemia inhibitory factor receptor inhibition reduces stenosis grade and total serum cholesterol levels in a mouse model for atherosclerosis
Authors: E Hemme, M A C Depuydt, P J Van Santbrink, L Delfos, J Kuiper, I Bot
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Dutch Heart Foundation Atherosclerosis is characterized by the accumulation of lipids and immune cells in the arterial wall and it is the main underlying pathology causing cardiovascular diseases (CVD). Cytokines are involved in all stages of atherosclerosis and most cytokines contribute to disease progression. For instance, IL-6 cytokine family members, such as IL-6, oncostatin-M and cardiotrophin-1, are closely related to CVDs. However, the role of leukemia inhibitory factor (LIF), a member of the IL-6 cytokine family, and its receptor (LIFR) in atherosclerosis remains unknown. Therefore, the aim of this study is to assess the contribution of LIFR signaling to atherosclerosis development by systemically inhibiting LIFR in an atherosclerotic mouse model. First, the expression of LIF and LIFR was examined using single-cell RNA sequencing data of human carotid artery plaques. Mast cells highly and exclusively express LIF, whereas LIFR was specifically expressed by activated endothelial cells. To validate LIFR signaling in vivo, female western-type diet fed Ldlr-/- mice were treated with LIFR inhibitor EC359 (5 mg/kg s.c., n=15) or control solvent (n=15) three times per week for eight weeks. During the experiment, weights of the mice did not differ between groups, whereas total serum cholesterol levels were significantly reduced in EC359 treated mice (p=0.03). In particular, EC359 treatment reduced VLDL levels. Additionally, EC359 treatment significantly reduced Sr1, Abca1 and Lrp1 expression in the liver, suggesting an altered lipid metabolism in EC359 treated mice. Furthermore, LIFR inhibition affected innate immunity, both systemically and locally in the plaque. Neutrophil numbers were significantly increased in the circulation (ctrl 18.7±1.7%; EC359 28.0±2.0%; p=0.002) and aortic plaques (ctrl 3.9±0.8%; EC359 9.9±1.9%; p=0.003) of mice treated with EC359 compared to control mice. Moreover, LIFR inhibition affected circulating monocytes resulting in less inflammatory monocytes. The adaptive immune cells, such as T and B cells, remained largely unaffected. Collectively, the reduction of serum total cholesterol and increase in neutrophil numbers in EC359 treated mice resulted in a trend towards reduced atherosclerotic stenosis grade compared to control mice (39.8±1.4% versus 34.9±1.9%; p=0.054), but it did not affect plaque collagen and monocyte/macrophage content in the aortic root. Conclusively, LIFR signaling inhibition reduces atherosclerotic stenosis grade, lowers plasma total cholesterol levels and affects innate immunity.
Cryopreservation of human adult ventricular tissue for the preparation of viable myocardial slices
Authors: A M Lodrini, A Husetic, T Kok, B P T Kruithof, J Hjortnaes, M Palmen, A M Smits, M J Goumans
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): REGeRNA Myocardial slices are ultra-thin slices of living adult myocardium which can be cultured. As a multicellular 3-D culture model, myocardial slices offer a novel opportunity to investigate how adult cardiomyocytes (CMs) interact with other cardiac cell types and the extracellular matrix, while providing long-term preservation of CMs cytoarchitecture and electrical phenotype. Additionally, myocardial slices are a new tool in drug discovery by allowing to test chronic treatments, as opposed to isolated CMs which are limited to acute timepoints. Despite their benefits over other cardiac preparations, myocardial slices also have several limitations, of which one is the quality of the slices. The main determinant of myocardial slice quality is the procedure by which the cardiac tissue block is handled. Current methods depend on the preparation of slices directly after tissue excision, therefore limiting the moment when the experiment is performed. We developed a protocol for the cryopreservation of human adult ventricular tissue allowing preparation of viable myocardial slices afterwards. Fresh and cryopreserved cardiac tissue from donor hearts was processed and evaluated for viability, tissue integrity, and inflammatory markers using molecular biology and histological assays. The tissue samples retained viability, biomass, and tissue and vascular integrity following cryopreservation compared to fresh tissue. Additionally, we compared myocardial slices obtained from fresh or cryopreserved tissue and cultured for 24 hours. Histologically, normal cytomorphological features and gap junctions between CMs were preserved in all slices. Comparable retention of tissue viability and structural integrity post cryopreservation (4-6 weeks) and culture (up to 7 days) was observed in cardiac samples from 3 additional donors. In summary, cold storage extends the time-interval between sample collection and slice preparation, allowing for more long-term, complex endpoints. Moreover, efficient banking of cryopreserved cardiac samples from various donors and disease states is critical for regular use of myocardial tissue slices in drug discovery and development. Additional research on the effects of cryopreservation on different cardiac cell types will allow to improve donor organ management to extend the donor pool and increase utilization rate.
OPA1 affects lipid metabolism and atherosclerosis progression
Authors: L Da Dalt, A Moregola, M Svecla, S Pedretti, F Fantini, E Donetti, N Mitro, L Scorrano, G D Norata
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): European Atherosclerosis Society Introduction Mitochondria are involved in cellular metabolism, energy generation, calcium homeostasis, sterol and bile acids (BAs) production. Mitochondria continuously undergo biogenesis, fusion, fission and mitophagy, maintaining a continuous balance between all forms. Purpose On these premises, we test the impact of OPA1, an inner mitochondria membrane fusion protein, on mitochondrial tethering on lipid metabolism in the liver and the atherosclerotic plaque. Methods OPA1HepKO and OPA1 TG on LDLR KO background were fed with a Western-type diet (WTD) respectively for 12 weeks. Inverse calorimetry, GTT, and LipidToleranceTest (LTT) were performed. Paraffin-embedded tissues were used for histological analysis, frozen tissues were used for OMICs analysis. VSMCs were isolated and cultured with VLDL (50 µg/ml). Results While OPA1HepKO mice display altered systemic metabolism with reduced body weight and reduced circulating lipid levels OPA1 overexpression on LDLR KO background showed significantly high cholesterol levels. OPA1 deficiency impair hepatic bile acid conjugation with significant accumulation of primary unconjugated bile acids in the liver. On the contrary OPA1 overexpression leads to a reduction of unconjugated bile acids and higher percentage of conjugated bile acids. OPA1 reduction leads to a reduced atherosclerotic plaque while, on the contrary, its overexpression doesn’t affect atherosclerosis despite an increase in circulating lipid and lipoprotein levels. OPA1 overexpression is therefore promoting plaque stabilization with VSMCs that are less prone to change their metabolism to synthetic. Conclusion Hepatic Opa1 deficiency alters bile acids production and therefore circulating lipid profile with a consequent reduction in atherosclerotic plaque formation. OPA1 overexpression in the aorta affects plaque stability despite the highest levels of circulating lipids.
Increased oxidative stress alters coronary microvascular tone in exercising swine with multiple comorbidities
Authors: R W A Van Drie, O Sorop, J Van De Wouw, D J Duncker, D Merkus
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Dutch Cardiovascular Alliance Background/Objectives We have previously shown, in female swine that the combination of three cardiovascular risk factors (diabetes mellitus (DM), hypercholestolemia (HC) and chronic kidney disease (CKD)) induces systemic inflammation, oxidative stress, coronary endothelial dysfunction and impaired myocardial perfusion due to impaired nitric oxide (NO) bioavailability. In the present study we hypothesized that the reduced bioavailability of nitric oxide (NO) is the result of increased production of reactive oxygen species (ROS) and ROS scavenging will restore myocardial oxygen balance thereby improving myocardial perfusion. Methods 12 female swine, with DM (streptozotocin 3x50mg/kg iv), CKD (renal embolization), and HC (high fat, high sugar, high salt diet) and 9 female healthy swine on normal pig chow (Normal) were included. The effect of ROS scavenging on the coronary microvascular tone control was studied in vivo, at rest and during graded treadmill exercise before and after infusion of the ROS scavengers (MPG+Tempol). Additionally, vasodilation to bradykinin was studied in the presence of MPG and TEMPOL and after subsequent addition to catalase, in isolated left ventricle small coronary arteries, in vitro using wire-myography. Results 6 months of DM+HC+CKD resulted in hyperglycemia (18.0±1.3 vs 8.4±0.9mmol/L), hypercholesterolemia (13.4±2.1 vs 1.7±0.1mmol/l), renal dysfunction (plasma creatinine: 166±9 vs 119±3µmol/l), and systemic inflammation (TNFα: 63±8 vs 41±4pg/ml, all P&amp;lt;0.05) associated with impaired endothelium-dependent vasodilator response to bradykinin in vitro. Furthermore, myocardial oxidative stress was increased in DM+HC+CKD animals (8-isoprostane 13±1 vs 10±1pg/ml) and total antioxidant capacity was reduced (0.67±0.02 vs 0.72±0.01nmol Trolox eq./mg protein, both P&amp;lt;0.05). Surprisingly, in vivo ROS scavenging resulted in an even further increase in myocardial oxygen extraction in DM+HC+CKD while it had no effect in Normal, indicating the involvement of a vasodilator ROS (most likely H2O2) in the control of coronary microvascular tone. This was supported by increased activity of catalase in the myocardium of DM+HC+CKD animals (44±3 vs 31±4nmol/min/ml, both P&amp;lt;0.05) and the vascular relaxation response to bradykinin when catalase was added on top of the ROS scavenging in vitro. Conclusion In swine exposed for 6 months to multiple comorbidities, impaired myocardial perfusion is mediated by a loss of NO bioavailability due to increased oxidative stress which was partially compensated by increased H2O2-mediated coronary vasodilation.
FHL2 is a novel target in pulmonary arterial hypertension
Authors: C Nadeem, A L C Abbas, D T B Verhoeven, L Willems, M J Goumans, K Kurakula
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): - Longfonds Nederland Background/ Objectives Pulmonary arterial hypertension (PAH) is a fatal disorder characterized by excessive proliferation of endothelial (EC) and smooth muscle cells, and impaired BMP signaling in small lung arteries. The LIM-only protein FHL2 is expressed in the vessel wall in ECs and smooth muscle cells and plays a crucial role in vessel wall homeostasis. However, the molecular mechanisms how FHL2 exerts its effects and its role in pulmonary artery EC function remain unclear. Here, we investigated the effect of FHL2 on EC proliferation and unraveled the underlying mechanisms involved. Methods/Results We found that FHL2 is highly expressed in lungs of PAH rats compared to control rats as evidenced by qPCR and western blot analysis. Human microvascular ECs (MVECs) derived from lungs of PAH patients displayed enhanced expression of FHL2. Ectopic expression of FHL2 in MVECs significantly increased mRNA levels of BMPR2 and Id1. To substantiate these results, luciferase assays were performed using BMP response element reporter construct and found that FHL2 increases Id1 expression. Furthermore, overexpression of FHL2 decreased TNF-α induced expression of pro-inflammatory cytokines. Finally, ectopic expression of FHL2 reduced proliferation of ECs. Conclusion Our data indicate that FHL2 is a novel negative regulator of MVEC proliferation through enhancing BMP signaling and inhibiting inflammation. Modulation of FHL2 may potentially represent a novel therapeutic strategy for the treatment of PAH.
MicroRNA-26b as novel regulator of cardio-metabolic diseases
Authors: E Van Der Vorst, L Rakateli, L Peters, K Bidzhekov, A Jans, S Maas, M Gijbels, J Jankowski, E Biessen, C Weber, Y Doering, M Bartneck
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): IZKF Increasing evidence has shown that microRNAs (miRs) are fundamental players in cardio-metabolic diseases. MiR-26b has been associated with the development of metabolic dysfunction-associated steatohepatitis (MASH) and atherosclerosis, although causality has not been demonstrated yet. Therefore, we aimed to determine the causal role of miR-26b in MASH and atherosclerosis. Apoe-/-Mir-26b-/- mice were fed a 4- or 12-week western type diet (WTD). Plasma and liver samples were subjected to various assays to investigate the effects on lipid metabolism, inflammation and fibrosis. Additionally, miR-26b mimic-loaded lipid nanoparticles (LNPs) were used to rescue the phenotype. A 1.5-fold increase in hepatic lipid levels was observed in Apoe-/-Mir26b-/- mice, while plasma cholesterol/triglyceride levels remained unchanged. These results were further confirmed by an increase in steatosis. Additionally, the proinflammatory cytokines TNF-α, IL-6 and CXCL1 were significantly elevated, which coincided with increased macrophage infiltration into the liver. Moreover, loss of miR-26b resulted in more hepatic fibrosis. Finally, a treatment with miR-26b mimic-loaded LNPs reduced hepatic lipid levels, rescuing the observed phenotype. Additionally, Apoe-/-Mir26b-/- mice demonstrated a striking 10-fold increase in atherosclerotic lesion size. Consistent with a more advanced plaque phenotype, collagen content and relative necrotic core area were significantly increased in plaques from mice lacking miR-26b. Intriguingly, this increased atherosclerotic lesion size could also be rescued by repetitive injections of miR-26b mimic-loaded LNPs. Overall, our results demonstrate a protective role of miR-26b in cardio-metabolic diseases, thereby opening doors to future research and potential new treatment options using LNP technology.
Cardiovascular toxicity profile of doxorubicin: data mining of the FDA adverse event reporting system (FAERS)
Authors: L Kattan, E Cerbito, H Dhulkifle, H Korashy, Z Maayah
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements None. Background Doxorubicin (DOX), an effective anticancer agent, is associated with dose-dependent cardiovascular toxicity. Understanding its mechanisms and risk factors will facilitate novel interventions to minimize DOX-induced cardiovascular toxicity. Purpose We aimed to perform a comprehensive analysis of the Food and Drug Administration Adverse Event Reporting System (FAERS) database to identify, describe, and quantify the latest cardiovascular DOX-associated adverse-events (AEs). Methods Data from 2004 Q1 – 2022 Q3, where DOX was the primary-suspect, were extracted from FAERS via OpenVigil 2.1. For data collection, preferred-terms (PTs) from Medical Dictionary for Regulatory Activities (MedDRA) were used, while ROR with 95% CIs for disproportionality analysis was performed. Results 1987 reports were collected for significant PTs. The most prevalent DOX-associated AE was heart failure (77.6%, n=1542), followed by cardiac fibrosis and inflammation (7.75%, n=154) and vascular disorders (7.45%, n=148). Others include arrhythmias, systemic inflammation, and endothelial dysfunction. Most patients were females (51.1%, n=1016), 18-64 years-old (41%, n = 815), primarily from Europe (41.87%, n=832) and the United States (34.12%, n = 678). Six outcomes were associated with DOX-induced cardiotoxicities, mainly "Death", "Initial/prolonged hospitalization", and "Others". Conclusion This study provides valuable insights on DOX-induced cardiovascular toxicity using real-world data from FAERS.Total Case Count of all Adverse EventPatients' Demographics and outcomes
Kbtbd13R408C-knockin mouse model reveals impaired relaxation kinetics as novel pathomechanism for NEM6 cardiomyopathy
Authors: R Baelde, V Janssen, A Fortes Monteiro, R Galli, M Methawasin, H Granzier, D Kuster, J Van Der Velden, C Ottenheijm, J De Winter
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): VENI-Nederlandse organisatie voor Wetenschappelijk Onderzoek (NWO) We identified a novel cardiomyopathy gene: KBTBD13 encoding kelch repeat and BTB (POZ) domain containing 13. The Dutch founder variant KBTBD13R408C not only affects skeletal muscle function resulting in nemaline myopathy type 6 (NEM6), but also affects the heart. In the Dutch NEM6 cohort, various cardiac abnormalities were observed, including systolic dysfunction, diastolic dysfunction, atrial fibrillation, ventricular tachycardia and three patients died of sudden cardiac death. The majority of NEM6 patients harbors the Dutch founder variant, c.1222C&amp;gt;T, p.Arg408Cys (KBTBD13R408C). To provide insight in the mechanism underlying cardiac dysfunction in NEM6, here we assessed cardiac structure and function in Kbtbd13R408C-knockin mice, which closely recapitulate the well characterized skeletal muscle phenotype, i.e. impaired relaxation kinetics and mitochondrial dysfunction. Pressure-volume loop analyses showed that the end-diastolic pressure-volume relation was steeper in Kbtbd13R408C-knockin mice, indicating diastolic dysfunction. Histological evaluation of cardiac structure revealed no increase in fibrosis in Kbtbd13R408C-knockin mice. Also, no alterations in titin isoform composition or myosin heavy chain composition were observed that could account for the increased diastolic stiffness. Next, we studied the contraction and relaxation kinetics in wild type (WT) and Kbtbd13R408C-knockin mice at the intact single cardiomyocyte level by a high-throughput contractility set-up. Our data showed a prolonged relaxation time in Kbtbd13R408C-knockin mice compared to WT mice, indicating impaired relaxation kinetics. No differences in the magnitude of contraction (percentage of shortening) was found between cardiomyocytes of Kbtbd13R408C-knockin and WT mice. In parallel, calcium-handling was assessed by calcium indicator Fura-2AM. These studies reveal that calcium-release is not affected, but calcium-reuptake is slower in Kbtbd13R408C-knockin mice, which might contribute to the impaired relaxation kinetics. Current studies focus on how KBTBD13R408C affects calcium-reuptake kinetics and sarcomere kinetics in NEM6 cardiomyocytes. Hence, our studies provide the first insights in the pathomechanism underlying cardiac dysfunction in NEM6.
Enhancing cellular regeneration through targeted pharmacological reprogramming of senescent endothelial cells
Authors: K Kalies, K Knoepp, L Wurmbrand, J Dutzmann, D Sedding
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements None. Background Senescent endothelial cells (EC) are key players in the pathophysiology of cardiovascular diseases and characterized by a reduced angiogenic and regenerative potential. Therefore, reversing EC senescence represents a promising therapeutic strategy to restore vascular integrity and increase health and lifespan. Purpose Here, we show a reversal of EC senescence following the application of a pharmacological, non-genetic, partial reprogramming strategy to induce a timely restricted induction of the Yamanaka-factors Oct3/4, Sox2, Klf4 and c-Myc (OSKM). Methods Methods to characterize the effects of pharmacological reprogramming included the quantification of gene expression as well as functional analysis of EC in vitro. In addition, the regenerative capacity of EC was evaluated in a hind-limb ischemia model in vivo in young and old C57BL/6 mice. Results The application of a pharmacological reprogramming cocktail led to a strong but timely restricted activation of the Yamanaka-factors Oct3/4, Sox2, Klf4 and c-Myc in replicative senescent ECs. This activation occurred on both mRNA and protein levels (p&amp;lt;0.0001), concurrently significantly reducing markers of cellular aging as p16ink4a, p14arf, TNFα, IL-1b, IL-6 and CD44 (p&amp;lt;0.01 respectively p&amp;lt;0.0001), while enhancing their cellular function. Pharmacological reprogramming notably augmented the migratory (p&amp;lt;0.001) and proliferative capacities (p&amp;lt;0.01) of the replicate senescence EC and positively influenced their angiogenic functions such as sprouting (p&amp;lt;0.0001) and tube formation (p&amp;lt;0.01). Telomere length was stabilized (p&amp;lt;0.05) and production of reactive oxygen species decreased (p&amp;lt;0.05). Non-senescent EC were not influenced by the pharmacological treatment (p&amp;gt;0.05). Over prolonged periods, the treated ECs maintained consistently low expression of aging markers (p16ink4a and p14arf, p&amp;lt;0.01) while preserving their migratory capabilities (p&amp;lt;0.05). In vivo, a significantly improved blood flow was observed after hind limb ischemia in 22 months old C57BL/6 mice after 7 and 14 days (p&amp;lt;0.001), alongside a significant increase in capillary density in the gastrocnemius muscle quantified by CD31 staining (p&amp;lt;0.05). In 3 months old C57BL/6 mice blood flow was not influenced by the treatment (p&amp;gt;0.05). Conclusion In conclusion, we demonstrate that a short induction of OSKM via a pharmacological approach holds the potential to reverse cellular senescence in EC in vitro with improved regenerative and angiogenic capacities and thus to enhance endothelial regenerative capacity in vivo. This might present a promising strategy for ischemic diseases.