ABSTRACT
BACKGROUND: Swine are frequently used as animal model for cardiovascular research, especially in terms of representativity of human anatomy and physiology. Reference values for the most common species used in research are important for planning and execution of animal testing. Transesophageal echocardiography is the gold standard for intraoperative imaging, but can be technically challenging in swine. Its predecessor, epicardial echocardiography (EE), is a simple and fast intraoperative imaging technique, which allows comprehensive and goal-directed assessment. However, there are few echocardiographic studies describing echocardiographic parameters in juvenile swine, none of them using EE. Therefore, in this study, we provide a comprehensive dataset on multiple geometric and functional echocardiographic parameters, as well as basic hemodynamic parameters in swine using EE. METHODS: The data collection was performed during animal testing in ten female swine (German Landrace, 104.4 ± 13.0 kg) before left ventricular assist device implantation. Hemodynamic data was recorded continuously, before and during EE. The herein described echocardiographic measurements were acquired according to a standardized protocol, encompassing apical, left ventricular short axis and long axis as well as epiaortic windows. In total, 50 echocardiographic parameters and 10 hemodynamic parameters were assessed. RESULTS: Epicardial echocardiography was successfully performed in all animals, with a median screening time of 14 min (interquartile range 11-18 min). Referring to left ventricular function, ejection fraction was 51.6 ± 5.9% and 51.2 ± 6.2% using the Teichholz and Simpson methods, respectively. Calculated ventricular mass was 301.1 ± 64.0 g, as the left ventricular end-systolic and end-diastolic diameters were 35.3 ± 2.5 mm and 48.2 ± 3.5 mm, respectively. The mean heart rate was 103 ± 28 bpm, mean arterial pressure was 101 ± 20 mmHg and mean flow at the common carotid artery was 627 ± 203 mL/min. CONCLUSION: Epicardial echocardiography allows comprehensive assessment of most common echocardiographic parameters. Compared to humans, there are important differences in swine with respect to ventricular mass, size and wall thickness, especially in the right heart. Most hemodynamic parameters were comparable between swine and humans. This data supports study planning, animal and device selection, reinforcing the three R principles in animal research.
Subject(s)
Echocardiography , Ventricular Function, Left , Humans , Female , Animals , Swine , Ventricular Function, Left/physiology , Echocardiography/methods , Hemodynamics , Heart Ventricles/diagnostic imagingABSTRACT
The new 2021 guidelines of the European Society of Cardiology (ESC) have broken with the old step by step treatment of heart failure and have fundamentally revised the strategy in the pharmacotherapy setting. For patients with heart failure and reduced ejection fraction ≤â¯40%, the 4 substance groups angiotensin-converting enzyme (ACE) inhibitors/angiotensin receptor-neprilysin inhibitors (ARNI), beta blockers, mineralocorticoid receptor antagonists (MRA) and sodium-glucose transporter 2 (SGLT2) inhibitors are now recommended as early as possible after diagnosis. Completing the substance groups has priority over increasing the dosage of the individual substances. This makes it necessary to rethink current clinical practice, especially as the guidelines are reluctant to give concrete instructions for implementation.
Subject(s)
Cardiology , Heart Failure , Ventricular Dysfunction, Left , Humans , Angiotensin Receptor Antagonists/therapeutic use , Heart Failure/drug therapy , Angiotensin-Converting Enzyme Inhibitors/therapeutic use , Ventricular Dysfunction, Left/drug therapy , Mineralocorticoid Receptor Antagonists/therapeutic use , Stroke VolumeABSTRACT
Dedifferentiated vascular smooth muscle cells (vSMCs) play an essential role in neointima formation, and we now aim to investigate the role of the bone morphogenetic protein (BMP) modulator BMPER (BMP endothelial cell precursor-derived regulator) in neointima formation. To assess BMPER expression in arterial restenosis, we used a mouse carotid ligation model with perivascular cuff placement. Overall BMPER expression after vessel injury was increased; however, expression in the tunica media was decreased compared to untreated control. Consistently, BMPER expression was decreased in proliferative, dedifferentiated vSMC in vitro. C57BL/6_Bmper+/- mice displayed increased neointima formation 21 days after carotid ligation and enhanced expression of Col3A1, MMP2, and MMP9. Silencing of BMPER increased the proliferation and migration capacity of primary vSMCs, as well as reduced contractibility and expression of contractile markers, whereas stimulation with recombinant BMPER protein had the opposite effect. Mechanistically, we showed that BMPER binds insulin-like growth factor-binding protein 4 (IGFBP4), resulting in the modulation of IGF signaling. Furthermore, perivascular application of recombinant BMPER protein prevented neointima formation and ECM deposition in C57BL/6N mice after carotid ligation. Our data demonstrate that BMPER stimulation causes a contractile vSMC phenotype and suggest that BMPER has the potential for a future therapeutic agent in occlusive cardiovascular diseases.
Subject(s)
Carrier Proteins , Neointima , Vascular Remodeling , Animals , Mice , Bone Morphogenetic Proteins/metabolism , Cell Movement , Cell Proliferation , Cells, Cultured , Disease Models, Animal , Mice, Inbred C57BL , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/metabolism , Neointima/metabolism , Phenotype , Carrier Proteins/metabolismABSTRACT
MicroRNAs are key regulators of the cardiac response to injury. MiR-100 has recently been suggested to be involved in different forms of heart failure, but functional studies are lacking. In the present study, we examined the impact of transgenic miR-100 overexpression on cardiac structure and function during physiological aging and pathological pressure-overload-induced heart failure in mice after transverse aortic constriction surgery. MiR-100 was moderately upregulated after induction of pressure overload in mice. While in our transgenic model the cardiomyocyte-specific overexpression of miR-100 did not result in an obvious cardiac phenotype in unchallenged mice, the transgenic mouse strain exhibited less left ventricular dilatation and a higher ejection fraction than wildtype animals, demonstrating an attenuation of maladaptive cardiac remodeling by miR-100. Cardiac transcriptome analysis identified a repression of several regulatory genes related to cardiac metabolism, lipid peroxidation, and production of reactive oxygen species (ROS) by miR-100 overexpression, possibly mediating the observed functional effects. While the modulation of ROS-production seemed to be indirectly affected by miR-100 via Alox5-and Nox4-downregulation, we demonstrated that miR-100 induced a direct repression of the scavenger protein CD36 in murine hearts resulting in a decreased uptake of long-chain fatty acids and an alteration of mitochondrial respiratory function with an enhanced glycolytic state. In summary, we identified miR-100 as a modulator of cardiac metabolism and ROS production without an apparent cardiac phenotype at baseline but a protective effect under conditions of pressure-overload-induced cardiac stress, providing new insight into the mechanisms of heart failure.
Subject(s)
CD36 Antigens/metabolism , Heart Failure/metabolism , MicroRNAs/metabolism , Myocytes, Cardiac/metabolism , NADPH Oxidase 4/metabolism , Reactive Oxygen Species/metabolism , Signal Transduction/genetics , Animals , CD36 Antigens/genetics , Disease Models, Animal , Fatty Acids/metabolism , HEK293 Cells , Heart Failure/genetics , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , MicroRNAs/genetics , NADPH Oxidase 4/genetics , Rats , Stroke Volume/genetics , Transfection , Ventricular Remodeling/geneticsABSTRACT
Aortic regurgitation (AR) following continuous flow left ventricular assist device implantation (cf-LVAD) may adversely impact outcomes. We aimed to assess the incidence and impact of progressive AR after cf-LVAD on prognosis, biomarkers, functional capacity and echocardiographic findings. In an analysis of the PCHF-VAD database encompassing 12 European heart failure centers, patients were dichotomized according to the progression of AR following LVAD implantation. Patients with de-novo AR or AR progression (AR_1) were compared to patients without worsening AR (AR_0). Among 396 patients (mean age 53 ± 12 years, 82% male), 153 (39%) experienced progression of AR over a median of 1.4 years on LVAD support. Before LVAD implantation, AR_1 patients were less frequently diabetic, had lower body mass indices and higher baseline NT-proBNP values. Progressive AR did not adversely impact mortality (26% in both groups, HR 0.91 [95% CI 0.61-1.36]; P = 0.65). No intergroup variability was observed in NT-proBNP values and 6-minute walk test results at index hospitalization discharge and at 6-month follow-up. However, AR_1 patients were more likely to remain in NYHA class III and had worse right ventricular function at 6-month follow-up. Lack of aortic valve opening was related to de-novo or worsening AR (P < 0.001), irrespective of systolic blood pressure (P = 0.67). Patients commonly experience de-novo or worsening AR when exposed to continuous flow of contemporary LVADs. While reducing effective forward flow, worsening AR did not influence survival. However, less complete functional recovery and worse RV performance among AR_1 patients were observed. Lack of aortic valve opening was associated with progressive AR.
Subject(s)
Aortic Valve Insufficiency , Heart Failure , Heart-Assist Devices , Humans , Male , Adult , Middle Aged , Aged , Female , Aortic Valve Insufficiency/diagnosis , Aortic Valve Insufficiency/epidemiology , Aortic Valve Insufficiency/etiology , Heart-Assist Devices/adverse effects , Heart Failure/diagnosis , Heart Failure/therapy , Heart Failure/complications , Echocardiography , Ventricular Function, Right , Retrospective Studies , Treatment OutcomeABSTRACT
BACKGROUND: Diet-induced obesity can result in the development of a diverse spectrum of cardiovascular and metabolic diseases, including type 2 diabetes, dyslipidemia, non-alcoholic liver steatosis and atherosclerotic disease. MicroRNAs have been described to be important regulators of metabolism and disease development. METHODS: In the current study, we investigated the effects of ubiquitous miR-100 overexpression on weight gain and the metabolic phenotype in a newly generated transgenic mouse strain under normal chow and high fat diet and used microarray expression analysis to identify new potential target genes of miR-100. RESULTS: While transgenic overexpression of miR-100 did not significantly affect weight and metabolism under a normal diet, miR-100 overexpressing mice showed a reduced weight gain under a high fat diet compared to wildtype mice, despite an equal calorie intake. This was accompanied by less visceral and subcutaneous fat development and lover serum LDL cholesterol. In addition, transgenic miR-100 mice were more glucose tolerant and insulin sensitive and demonstrated increased energy expenditure under high fat diet feeding. A comprehensive gene expression profiling revealed the differential expression of several genes involved in lipid storage- and metabolism, among them CD36 and Cyp4A14. Our data showed a direct regulation of CD36 by miR-100, leading to a reduced fatty acid uptake in primary hepatocytes overexpressing miR-100 and the downregulation of several downstream mediators of lipid metabolism such as ACC1, FABP4, FAS and PPARγ in the liver. CONCLUSIONS: Our findings demonstrate a protective role of miR-100 in high fat diet induced metabolic syndrome and liver steatosis, partially mediated by the direct repression of CD36 and attenuation of hepatic lipid storage, implicating miR-100 as a possible therapeutic target in liver steatosis.
Subject(s)
Hypertriglyceridemia/etiology , Hypertriglyceridemia/metabolism , Metabolic Syndrome/etiology , Metabolic Syndrome/metabolism , MicroRNAs/genetics , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/metabolism , 3' Untranslated Regions , Animals , Biomarkers , Cells, Cultured , Diet, High-Fat/adverse effects , Disease Models, Animal , Disease Susceptibility , Gene Expression Regulation , Glucose/metabolism , Hepatocytes/metabolism , Insulin/metabolism , Lipid Metabolism , Male , Mice , Mice, Transgenic , Phenotype , RNA Interference , Transcriptome , Weight GainABSTRACT
In the coronavirus disease 2019 (COVID-19) pandemic, organ transplant recipients are considered to be at high risk for an unfavorable outcome. However, in particular the role of immunosuppression in patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains undetermined. Here, we present a 62-year-old male COVID-19 patient with recent heart transplantation who developed only mild symptoms, but had prolonged virus shedding, and summarize the available data on COVID-19 in cardiac allograft recipients. Initially the patient presented with a transient episode of fever and sore throat but no other symptoms, in particular no cough or dyspnea at rest. After diagnosis, immunosuppression was continued unchanged. On day 7, his temperature increased again with concurrent mild rise of C-reactive protein, IL-6, and pro-B-type natriuretic peptide levels. Hydroxychloroquine was started and continued for 7 days. While the patient no longer had clinical symptoms 20 days after initial presentation, virus culture of throat swabs on days 18 and 21 confirmed active virus replication and SARS-CoV-2 PCR remained positive on day 35 with copy numbers similar to the onset of infection. In conclusion, the immunosuppression regimen in transplant recipients with mild COVID-19-associated symptoms may be continued unchanged. However, it may contribute to delayed virus polymerase chain reaction conversion and thus possible prolonged infectivity.
Subject(s)
COVID-19/epidemiology , Heart Failure/surgery , Heart Transplantation/methods , RNA, Viral/analysis , SARS-CoV-2/genetics , Virus Shedding , Aged , Antiviral Agents/therapeutic use , COVID-19/virology , Comorbidity , Heart Failure/epidemiology , Humans , Male , Middle Aged , Pandemics , Transplant Recipients , COVID-19 Drug TreatmentABSTRACT
BACKGROUND: In patients with chronic heart failure (CHF), volume overload is usually described as an expansion of plasma volume. Additional red cell volume (RCV) expansion is less commonly recognized. So far, little is known about quantitative differences in blood volume status and its different components in patients with stable CHF compared to healthy controls. METHODS: This study aimed to quantify blood volume and its constituents, RCV and plasma volume, by using an abbreviated carbon monoxide rebreathing method with particular focus on its primary measure total hemoglobin mass in 47 patients (10 women) with systolic CHF and a left ventricular ejection fraction of 29.0 ± 9.4%. These were compared to an age-matched control group of 84 healthy subjects (44 women) using the same method. RESULTS: In both absolute and body-surface-area-corrected analysis, hemoglobin mass (446 ± 81 vs 353 ± 64 g/m2) as well as RCV (1293 ± 231 vs 1033 ± 176 mL/m2) were significantly increased in CHF. In addition, significant plasma volume expansion was observed in CHF (2069 ± 400 vs 1750 ± 231 mL/m2) and, in conjunction with RCV, constituted a significantly increased blood volume (3361 ± 574 vs 2783 ± 369 mL/m2). In 66% of patients with compensated CHF, RCV was excessive compared to 14% in the control group. CONCLUSIONS: An increased RCV is a relevant contributing factor to hypervolemia in stable CHF. This is associated with an increased oxygen-carrying capacity, so it may be regarded as a possible compensatory mechanism for a reduced ejection fraction.
Subject(s)
Heart Failure, Systolic , Heart Failure , Blood Volume , Cell Size , Female , Heart Failure, Systolic/diagnosis , Humans , Stroke Volume , Ventricular Function, LeftABSTRACT
RATIONALE: The interaction of circulating cells within the vascular wall is a critical event in chronic inflammatory processes, such as atherosclerosis, but the control of the vascular inflammatory state is still largely unclear. OBJECTIVE: This study was undertaken to characterize the function of the endothelial-enriched microRNA miR-100 during vascular inflammation and atherogenesis. METHODS AND RESULTS: Based on a transcriptome analysis of endothelial cells after miR-100 overexpression, we identified miR-100 as a potent suppressor of endothelial adhesion molecule expression, resulting in attenuated leukocyte-endothelial interaction in vitro and in vivo as shown by flow cytometry and intravital imaging. Mechanistically, miR-100 directly repressed several components of mammalian target of rapamycin complex 1-signaling, including mammalian target of rapamycin and raptor, which resulted in a stimulation of endothelial autophagy and attenuated nuclear factor κB signaling in vitro and in vivo. In a low-density lipoprotein receptor-deficient atherosclerotic mouse model, pharmacological inhibition of miR-100 resulted in enhanced plaque lesion formation and a higher macrophage content of the plaque, whereas a systemic miR-100 replacement therapy had protective effects and attenuated atherogenesis, resulting in a decrease of plaque area by 45%. Finally, analysis of miR-100 expression in >70 samples obtained during carotid endarterectomy revealed that local miR-100 expression was inversely correlated with inflammatory cell content in patients. CONCLUSIONS: In summary, we describe an anti-inflammatory function of miR-100 in the vascular response to injury and inflammation and identify an important novel modulator of mammalian target of rapamycin signaling and autophagy in the vascular system. Our findings of miR-100 as a potential protective anti-athero-miR suggest that the therapeutic replacement of this microRNA could be a potential strategy for the treatment of chronic inflammatory diseases, such as atherosclerosis, in the future.
Subject(s)
Atherosclerosis/pathology , Autophagy , Endothelial Cells/pathology , MicroRNAs/physiology , Vasculitis/pathology , Animals , Carotid Artery Diseases/metabolism , Cell Adhesion Molecules/biosynthesis , Cell Adhesion Molecules/genetics , Cholesterol, LDL/metabolism , Human Umbilical Vein Endothelial Cells , Humans , Leukocytes/physiology , Macrophages/physiology , Male , Mice , Mice, Inbred C57BL , Receptors, LDL/metabolism , Simvastatin/pharmacology , Specific Pathogen-Free Organisms , TOR Serine-Threonine Kinases/physiology , TranscriptomeABSTRACT
BACKGROUND: Increased aortic stiffness is an independent predictor of cardiovascular disease. Optimal measurement is highly beneficial for the detection of atherosclerosis and the management of patients at risk. Thus, it was our purpose to selectively measure aortic stiffness using a novel imaging method and to provide reference values from a population-based study. METHODS: One hundred twenty six inhabitants of Freiburg, Germany, between 20 and 80 years prospectively underwent 3 Tesla cardiovascular magnetic resonance (CMR) of the thoracic aorta. 4D flow CMR (spatial/temporal resolution 2mm3/20ms) was executed to calculate aortic pulse wave velocity (PWV) in m/s using dedicated software. In addition, we calculated distensibility coefficients (DC) using 2D CINE CMR imaging of the ascending (AAo) and descending aorta (DAo). Segmental aortic diameter and thickness of aortic plaques were determined by 3D T1 weighted CMR (spatial resolution 1mm3). RESULTS: PWV increased from 4.93 ± 0.54 m/s in 20-30 year-old to 8.06 ± 1.03 m/s in 70-80 year-old subjects. PWV was significantly lower in women compared to men (p < 0.0001). Increased blood pressure (systolic r = 0.36, p < 0.0001; diastolic r = 0.33, p = 0.0001; mean arterial pressure r = 0.37, p < 0.0001) correlated with PWV after adjustment for age and gender. Finally, PWV increased with increasing diameter of the aorta (ascending aorta r = 0.20, p = 0.026; aortic arch r = 0.24, p = 0.009; descending aorta r = 0.26, p = 0.004). Correlation of PWV and DC of the AAo and DAo or the mean of both was high (r = 0.69, r = 0.68, r = 0.73; p < 0.001). CONCLUSIONS: 4D flow CMR was successfully applied to calculate aortic PWV and thus aortic stiffness. Findings showed a high correlation with distensibility coefficients representing local compliance of the aorta. Our novel method and reference data for PWV may provide a reliable biomarker for the identification of patients with underlying cardiovascular disease and optimal guidance of future treatment in studies or clinical routine.
Subject(s)
Aorta, Thoracic/diagnostic imaging , Aortic Diseases/diagnostic imaging , Magnetic Resonance Angiography/methods , Magnetic Resonance Imaging, Cine/methods , Vascular Stiffness , Adult , Age Factors , Aged , Aged, 80 and over , Aorta, Thoracic/pathology , Aorta, Thoracic/physiopathology , Aortic Diseases/epidemiology , Aortic Diseases/pathology , Aortic Diseases/physiopathology , Blood Flow Velocity , Cross-Sectional Studies , Female , Germany/epidemiology , Humans , Incidence , Male , Middle Aged , Plaque, Atherosclerotic , Predictive Value of Tests , Prospective Studies , Pulse Wave Analysis , Regional Blood Flow , Risk Factors , Sex Factors , Young AdultABSTRACT
BACKGROUND: To assess age-related changes of blood flow and geometry of the caval veins and right atrium (RA) using 4D flow cardiovascular magnetic resonance (CMR) data obtained in a population-based study. METHODS: An age-stratified sample (n = 126) of the population of the city of Freiburg, Germany, underwent transthoracic echocardiography and electrocardiogram-triggered and navigator-gated 4D flow CMR at 3 Tesla covering the caval veins and right heart. Study participants were divided into three age groups (1:20-39; 2:40-59; and 3:60-80 years of age). Analysis planes were placed in the superior and inferior caval vein. Subsequently, RA morphology and three-dimensional blood inflow pattern was assessed. RESULTS: Blood flow of the RA showed a clockwise rotating helix without signs of turbulence in younger subjects. By contrast, such rotation was absent in 12 subjects of group 3 and turbulences were significantly more frequent (p < 0.001). We observed an age-related shift of the caval vein axis. While the outlets of the superior and inferior caval veins were facing each other in group 1, lateralization occurred in older subjects (p < 0.001). A convergence of axes was observed from lateral view with facing axes in older subjects (p = 0.004). Finally, mean and peak systolic blood flow in the caval veins decreased with age (group 3 < 2 < 1). CONCLUSIONS: We have provided reference values of 4D CMR blood flow for different age groups and demonstrated the significant impact of age on hemodynamics of the RA inflow tract. This effect of aging should be taken into account when assessing pathologic conditions of the heart in the future.
Subject(s)
Aging , Heart Atria/diagnostic imaging , Image Interpretation, Computer-Assisted/methods , Magnetic Resonance Imaging/methods , Myocardial Perfusion Imaging/methods , Vena Cava, Inferior/diagnostic imaging , Vena Cava, Superior/diagnostic imaging , Adult , Age Factors , Aged , Aged, 80 and over , Atrial Function, Right , Blood Flow Velocity , Cardiac-Gated Imaging Techniques , Cross-Sectional Studies , Echocardiography , Electrocardiography , Germany , Heart Atria/physiopathology , Humans , Middle Aged , Predictive Value of Tests , Risk Factors , Vena Cava, Inferior/physiopathology , Vena Cava, Superior/physiopathology , Young AdultABSTRACT
The successful treatment of acute leukemias with allogeneic hematopoietic cell transplantation (allo-HCT) is limited by acute graft-versus-host disease (GVHD). Because microRNA-155 (miR-155) regulates activation of the innate immune system, we aimed to determine its function in dendritic cells (DCs) during GVHD in an experimental model. We observed that miR-155 deficiency of the recipient led to improved survival, reduced serum levels of proinflammatory cytokines, and lower GVHD histopathology scores. In addition, miR-155(-/-) bone marrow chimeric mice receiving allo-HCT and miR-155(-/-) DCs showed that miR-155 deficiency in the DC compartment was responsible for protection from GVHD. Activated miR-155(-/-) DCs displayed lower expression of various purinergic receptors and impaired migration toward adenosine triphosphate (ATP). Microarray analysis of lipopolysaccharide/ATP-stimulated miR-155(-/-) DCs revealed mitogen-activated protein kinase pathway dysregulation and reduced inflammasome-associated gene expression. Consistent with this gene expression data, we observed reduced ERK activation, caspase-1 cleavage, and IL-1ß production in miR-155(-/-) DCs. The connection between miR-155 and inflammasome activation was supported by the fact that Nlrp3/miR-155 double-knockout allo-HCT recipient mice had no increased protection from GVHD compared with Nlrp3(-/-) recipients. This study indicates that during GVHD, miR-155 promotes DC migration toward sites of ATP release accompanied by inflammasome activation. Inhibiting proinflammatory miR-155 by antagomir treatment could help reduce this complication of allo-HCT.
Subject(s)
Cell Movement , Dendritic Cells/immunology , Graft vs Host Disease/genetics , Inflammasomes/metabolism , MicroRNAs/genetics , Animals , Cell Movement/genetics , Cell Movement/immunology , Cells, Cultured , Dendritic Cells/physiology , Female , Graft vs Host Disease/immunology , Hematopoietic Stem Cell Transplantation/adverse effects , Inflammasomes/genetics , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , MicroRNAs/metabolism , Severity of Illness Index , Transplantation, Homologous/adverse effectsABSTRACT
Background Global longitudinal strain is reduced in heart transplant recipients, but little is known about regional contractility of the transplanted heart. Moreover, it is unclear if factors such as time after transplant and ischemic time have an influence on regional contractility. To test for regional changes in myocardial deformation, we assessed regional myocardial deformation using three-dimensional speckle tracking echocardiography in heart transplant recipients and controls. Methods Global and regional longitudinal, circumferential, and radial strain was assessed in 51 heart transplant recipients and a control group of healthy individuals (n = 26). Moreover, we correlated regional contractility with clinical characteristics and compared subgroups of heart transplant recipients with normal (n = 32) and reduced left ventricular ejection fraction (n = 32). Results Global longitudinal and circumferential strain was significantly reduced in all heart transplant recipients, as well as in the transplant group with normal ejection fraction compared with the control group (p < 0.001). Global radial strain (GRS) was elevated in both transplant groups, but was significantly higher in transplant recipients with normal ejection fraction compared with the control group (p < 0.01). Both transplant groups revealed lower longitudinal and circumferential strain values in the mid- and apical regions (p < 0.001), whereas longitudinal and circumferential strain was higher in the basal region (p < 0.01). In both groups, transplanted hearts showed increased radial strain in the basal (p < 0.05, p < 0.01) and midregions (p = 0.22; p < 0.01) and did not differ in the apical regions. Cold ischemic time (150 ± 12.6 minutes) was independently associated with reduction in circumferential strain. Time since transplantation ranging from 18 days to 21 years had no effect on myocardial deformation parameters. Conclusion Left ventricular mechanics in transplanted hearts display significantly different systolic deformation patterns than native hearts. Strain capability forms a regional gradient from the base toward the apex. The presence of a time-independent deformation pattern and the correlation with cold ischemic time suggest damage induced by the transplantation itself. These findings might be important for understanding pseudo-abnormal echocardiograms in heart transplant patients.
Subject(s)
Cold Ischemia/adverse effects , Heart Transplantation/adverse effects , Myocardial Contraction , Myocardial Reperfusion Injury/etiology , Ventricular Dysfunction, Left/etiology , Ventricular Function, Left , Adult , Aged , Biomechanical Phenomena , Echocardiography, Three-Dimensional , Female , Heart Transplantation/methods , Humans , Longitudinal Studies , Male , Middle Aged , Myocardial Reperfusion Injury/diagnostic imaging , Myocardial Reperfusion Injury/physiopathology , Predictive Value of Tests , Retrospective Studies , Risk Factors , Stress, Mechanical , Stroke Volume , Time Factors , Treatment Outcome , Ventricular Dysfunction, Left/diagnostic imaging , Ventricular Dysfunction, Left/physiopathologyABSTRACT
BACKGROUND: Adaptive neovascularization after arterial occlusion is an important compensatory mechanism in cardiovascular disease and includes both the remodeling of pre-existing vessels to collateral arteries (arteriogenesis) and angiogenic capillary growth. We now aimed to identify regulatory microRNAs involved in the modulation of neovascularization after femoral artery occlusion in mice. METHODS AND RESULTS: Using microRNA-transcriptome analysis, we identified miR-155 as a downregulated microRNA during hindlimb ischemia. Correspondingly, inhibition of miR-155 in endothelial cells had a stimulatory effect on proliferation and angiogenic tube formation via derepression of its direct target gene angiotensin II type 1 receptor. Surprisingly, miR-155-deficient mice showed an unexpected phenotype in vivo, with a strong reduction of blood flow recovery after femoral artery ligation (arteriogenesis) dependent on the attenuation of leukocyte-endothelial interaction and a reduction of proarteriogenic cytokine expression. Consistently, miR-155-deficient macrophages exhibit a specific alteration of the proarteriogenic cytokine expression profile, which is partly mediated by the direct miR-155 target gene SOCS-1. CONCLUSIONS: Our data demonstrate that miR-155 exerts an antiangiogenic but proarteriogenic function in the regulation of neovascularization via the suppression of divergent cell-specific target genes and that its expression in both endothelial and bone marrow-derived cells is essential for arteriogenesis in response to hindlimb ischemia in mice.
Subject(s)
Collateral Circulation/genetics , Hindlimb/blood supply , Ischemia/genetics , MicroRNAs/physiology , Neovascularization, Physiologic/genetics , Animals , Arteries/physiopathology , Base Sequence , Cell Movement , Cytokines/physiology , Down-Regulation , Endothelium, Vascular/physiopathology , Femoral Artery , Gene Expression Regulation , Human Umbilical Vein Endothelial Cells , Intercellular Signaling Peptides and Proteins/physiology , Laser-Doppler Flowmetry , Leukocytes/physiology , Ligation , Macrophages/physiology , Mice , Mice, Inbred C57BL , Mice, Knockout , MicroRNAs/genetics , Molecular Sequence Data , Receptor, Angiotensin, Type 1/biosynthesis , Receptor, Angiotensin, Type 1/genetics , Receptor, Angiotensin, Type 1/physiology , Suppressor of Cytokine Signaling 1 Protein , Suppressor of Cytokine Signaling Proteins/physiologyABSTRACT
BACKGROUND: Whole body ischemia-reperfusion injury (IRI) after cardiopulmonary resuscitation (CPR) induces a generalized inflammatory response which contributes to the development of post-cardiac arrest syndrome (PCAS). Recently, pattern recognition receptors (PRRs), such as toll-like receptors (TLRs) and inflammasomes, have been shown to mediate the inflammatory response in IRI. In this study we investigated monocyte PRR signaling and function in PCAS. METHODS: Blood samples were drawn in the first 12 hours, and at 24 and 48 hours following return of spontaneous circulation in 51 survivors after cardiac arrest. Monocyte mRNA levels of TLR2, TLR4, interleukin-1 receptor-associated kinase (IRAK)3, IRAK4, NLR family pyrin domain containing (NLRP)1, NLRP3, AIM2, PYCARD, CASP1, and IL1B were determined by real-time quantitative PCR. Ex vivo cytokine production in response to stimulation with TLR ligands Pam3CSK4 and lipopolysaccharide (LPS) was assessed in both whole blood and monocyte culture assays. Ex vivo cytokine production of peripheral blood mononuclear cells (PBMCs) from a healthy volunteer in response to stimulation with patients' sera with or without LPS was assessed. The results were compared to 19 hemodynamically stable patients with coronary artery disease. RESULTS: Monocyte TLR2, TLR4, IRAK3, IRAK4, NLRP3, PYCARD and IL1B were initially upregulated in patients following cardiac arrest. The NLRP1 and AIM2 inflammasomes were downregulated in resuscitated patients. There was a significant positive correlation between TLR2, TLR4, IRAK3 and IRAK4 expression and the degree of ischemia as assessed by serum lactate levels and the time until return of spontaneous circulation. Nonsurvivors at 30 days had significantly lower mRNA levels of TLR2, IRAK3, IRAK4, NLRP3 and CASP1 in the late phase following cardiac arrest. We observed reduced proinflammatory cytokine release in response to both TLR2 and TLR4 activation in whole blood and monocyte culture assays in patients after CPR. Sera from resuscitated patients attenuated the inflammatory response in cultured PBMCs after co-stimulation with LPS. CONCLUSIONS: Successful resuscitation from cardiac arrest results in changes in monocyte pattern recognition receptor signaling pathways, which may contribute to the post-cardiac arrest syndrome. TRIAL REGISTRATION: The trial was registered in the German Clinical Trials Register ( DRKS00009684 ) on 27/11/2015.
Subject(s)
Cardiopulmonary Resuscitation/mortality , Inflammasomes/pharmacokinetics , Signal Transduction/physiology , Toll-Like Receptors/metabolism , Adaptor Proteins, Signal Transducing/analysis , Adaptor Proteins, Signal Transducing/blood , Aged , Apoptosis Regulatory Proteins/analysis , Apoptosis Regulatory Proteins/blood , CARD Signaling Adaptor Proteins , Cytoskeletal Proteins/analysis , Cytoskeletal Proteins/blood , DNA-Binding Proteins/analysis , DNA-Binding Proteins/blood , Female , Germany , Homeodomain Proteins/analysis , Homeodomain Proteins/blood , Humans , Interleukin-1 Receptor-Associated Kinases/analysis , Interleukin-1 Receptor-Associated Kinases/blood , Interleukin-1beta/analysis , Interleukin-1beta/blood , Male , Middle Aged , Monocytes/chemistry , Monocytes/metabolism , Monocytes/pathology , NLR Family, Pyrin Domain-Containing 3 Protein/analysis , NLR Family, Pyrin Domain-Containing 3 Protein/blood , NLR Proteins , Nuclear Proteins/analysis , Nuclear Proteins/blood , Prospective Studies , Reperfusion Injury/complications , Reperfusion Injury/etiology , Repressor Proteins/analysis , Repressor Proteins/blood , Systemic Inflammatory Response Syndrome/prevention & control , Toll-Like Receptor 2/analysis , Toll-Like Receptor 2/blood , Toll-Like Receptor 4/analysis , Toll-Like Receptor 4/blood , Transcription FactorsABSTRACT
Percutaneous pulmonary valve implantation (PPVI) has become a widely practiced treatment option for patients with right ventricular outflow tract (RVOT) dysfunction. For this procedure, the Melody transcatheter pulmonary valve has become the most widely used system. On-label indications are limited to the treatment of failing circumferential conduits. However, there is growing but still limited experience with the off-label treatment of native or patched RVOTs. Here, the case is reported of a 57-year-old patient in whom redo-surgery for a dysfunctional pulmonary valve could be avoided by PPVI into the patched RVOT at 51 years after tetralogy of Fallot repair.
Subject(s)
Heart Valve Prosthesis Implantation/methods , Pulmonary Valve Stenosis/surgery , Pulmonary Valve/surgery , Tetralogy of Fallot/surgery , Ventricular Outflow Obstruction/surgery , Age Factors , Female , Humans , Middle Aged , Time Factors , Treatment OutcomeABSTRACT
Arteriogenesis is a complicated process induced by increased local shear-and radial wall-stress, leading to an increase in arterial diameter. This process is enhanced by growth factors secreted by both inflammatory and endothelial cells in response to physical stress. Although therapeutic promotion of arteriogenesis is of great interest for ischaemic diseases, little is known about the modulation of the signalling cascades via microRNAs. We observed that miR-132/212 expression was significantly upregulated after occlusion of the femoral artery. miR-132/212 knockout (KO) mice display a slower perfusion recovery after hind-limb ischaemia compared to wildtype (WT) mice. Immunohistochemical analysis demonstrates a clear trend towards smaller collateral arteries in KO mice. Although Ex vivo aortic ring assays score similar number of branches in miR-132/212 KO mice compared to WT, it can be stimulated with exogenous miR-132, a dominant member of the miR-132/212 family. Moreover, in in vitro pericyte-endothelial co-culture cell assays, overexpression of miR-132 and mir-212 in endothelial cells results in enhanced vascularization, as shown by an increase in tubular structures and junctions. Our results suggested that miR-132/212 may exert their effects by enhancing the Ras-Mitogen-activated protein kinases MAPK signalling pathway through direct inhibition of Rasa1, and Spred1. The miR-132/212 cluster promotes arteriogenesis by modulating Ras-MAPK signalling via direct targeting of its inhibitors Rasa1 and Spred1.
Subject(s)
Hindlimb/blood supply , Ischemia/genetics , MicroRNAs/metabolism , Mitogen-Activated Protein Kinases/metabolism , Morphogenesis , ras Proteins/metabolism , Animals , Gene Knockdown Techniques , HEK293 Cells , Hindlimb/drug effects , Hindlimb/pathology , Human Umbilical Vein Endothelial Cells , Humans , In Vitro Techniques , Intercellular Signaling Peptides and Proteins/pharmacology , Ischemia/pathology , MAP Kinase Signaling System/drug effects , Mice, Inbred C57BL , Mice, Knockout , MicroRNAs/genetics , Morphogenesis/drug effects , Neovascularization, Physiologic/drug effects , Phosphorylation/drug effects , Up-Regulation/drug effectsABSTRACT
The inflammatory sequelae of ischemia-reperfusion injury (IRI) are a major causal factor of tissue injury in various clinical settings. MicroRNAs (miRs) are short, non-coding RNAs, which regulate protein expression. Here, we investigated the role of miR-155 in IR-related tissue injury. Quantifying microRNA-expression levels in a human muscle tissue after IRI, we found miR-155 expression to be significantly increased and to correlate with the increased expression of TNF-α, IL-1ß, CD105, and Caspase3 as well as with leukocyte infiltration. The direct miR-155 target gene SOCS-1 was downregulated. In a mouse model of myocardial infarction, temporary LAD ligation and reperfusion injury resulted in a smaller area of necrosis in miR-155-/- animals compared to wildtype animals. To investigate the underlying mechanisms, we evaluated the effect of miR-155 on inflammatory cell recruitment by intravital microscopy and on the generation of reactive oxygen species (ROS) of macrophages. Our intravital imaging results demonstrated a decreased recruitment of inflammatory cells in miR-155-/- animals during IRI. The generation of ROS in leukocytic cells of miR-155-/- animals was also reduced. RNA silencing of the direct miR-155 target gene SOCS-1 abrogated this effect. In conclusion, miR-155 aggravates the inflammatory response, leukocyte infiltration and tissue damage in IRI via modulation of SOCS-1-dependent generation of ROS. MiR-155 is thus a potential target for the treatment or prevention of IRI.
Subject(s)
Chemotaxis, Leukocyte/physiology , Inflammation/metabolism , MicroRNAs/metabolism , Reperfusion Injury/metabolism , Transendothelial and Transepithelial Migration/physiology , Animals , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Humans , Inflammation/genetics , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Real-Time Polymerase Chain Reaction , Reperfusion Injury/genetics , Respiratory Burst/physiology , Suppressor of Cytokine Signaling 1 Protein , Suppressor of Cytokine Signaling Proteins/metabolism , TransfectionABSTRACT
Acute graft-versus-host disease (GvHD) is a complex process involving endothelial damage and neovascularization. Better understanding of the pathophysiology of neovascularization during GvHD could help to target this process while leaving T-cell function intact. Under ischemic conditions, neovascularization is regulated by different micro RNAs (miRs), which potentially play a role in inflamed hypoxic GvHD target organs. We observed strong neovascularization in the murine inflamed intestinal tract (IT) during GvHD. Positron emission tomography imaging demonstrated abundant αvß3 integrin expression within intestinal neovascularization areas. To interfere with neovascularization, we targeted αv integrin-expressing endothelial cells, which blocked their accumulation in the IT and reduced GvHD severity independent of immune reconstitution and graft-versus-tumor effects. Additionally, enhanced neovascularization and αv integrin expression correlated with GvHD severity in humans. Expression analysis of miRs in the inflamed IT of mice developing GvHD identified miR-100 as significantly downregulated. Inactivation of miR-100 enhanced GvHD indicating a protective role for miR-100 via blocking inflammatory neovascularization. Our data from the mouse model and patients indicate that inflammatory neovascularization is a central event during intestinal GvHD that can be inhibited by targeting αv integrin. We identify negative regulation of GvHD-related neovascularization by miR-100, which indicates common pathomechanistic features of GvHD and ischemia.
Subject(s)
Graft vs Host Disease/complications , Inflammation/etiology , Integrin alphaV/metabolism , Intestinal Diseases/etiology , MicroRNAs/genetics , Neovascularization, Pathologic , Animals , Blotting, Western , Bone Marrow Transplantation , Female , Flow Cytometry , Graft vs Host Disease/metabolism , Graft vs Host Disease/pathology , Humans , Immunoenzyme Techniques , Inflammation/metabolism , Inflammation/pathology , Integrin alphaV/genetics , Intestinal Diseases/metabolism , Intestinal Diseases/pathology , Luminescent Measurements , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , MicroRNAs/metabolism , Positron-Emission Tomography , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Epithelial injury is a central finding in pulmonary disease and is accompanied by disruption of epithelial barrier function, leading to pulmonary oedema and inflammation. Injured epithelial cells lose their properties and gain mesenchymal characteristics, a phenotypic switch that contributes to lung remodelling after injury. Here we studied bone morphogenetic protein (BMP) signalling and, in particular, the role of BMP2 and the BMP modulator BMPER in injured lung epithelium. Increased BMP activity, reflected by up-regulation of the Smad1/5-Id1 axis, is detected after injury of lung epithelium in vitro and in vivo. Two members of the BMP family, BMP2 and BMPER, have opposing effects. BMP2 is up-regulated after epithelial injury and causes epithelial dysfunction and hyperpermeability, mediated by the Smad1/5-Id1-dependent down-regulation of E-cadherin. In contrast, BMPER expression is decreased following injury, which in turn impairs epithelial integrity, characterized by reduction of E-cadherin and epithelial leakage in vitro and in vivo. High levels of BMPER antagonized BMP2-Smad5-Id1 signalling and prevented BMP2-mediated decrease of E-cadherin and hyperpermeability, suggesting that BMPER restores epithelial homeostasis. Supporting this notion, pharmacological inhibition of BMP signalling by LDN193189 prevented reduction of E-cadherin and disruption of epithelial barrier function. Inhibition of excessive BMP activation could be a new approach to restore epithelial integrity and prevent disruption of epithelial barrier function after lung injury.