Plasma Extracellular Vesicles as Liquid Biopsy to Unravel the Molecular Mechanisms of Cardiac Reverse Remodeling Following Resynchronization Therapy?

Cardiac resynchronization therapy (CRT) has become a valuable addition to the treatment options for heart failure, in particular for patients with disturbances in electrical conduction that lead to regionally different contraction patterns (dyssynchrony). Dyssynchronous hearts show extensive molecular and cellular remodeling, which has primarily been investigated in experimental animals. Evidence showing that at least several miRNAs play a role in this remodeling is increasing. A comparison of results from measurements in plasma and myocardial tissue suggests that plasma levels of miRNAs may reflect the expression of these miRNAs in the heart. Because many miRNAs released in the plasma are included in extracellular vesicles (EVs), which protect them from degradation, measurement of myocardium-derived miRNAs in peripheral blood EVs may open new avenues to investigate and monitor (reverse) remodeling in dyssynchronous and resynchronized hearts of patients.

Left atrial remodeling in mitral regurgitation: A combined experimental-computational study.

AIMS: Progressive changes to left atrial (LA) structure and function following mitral regurgitation (MR) remain incompletely understood. This study aimed to demonstrate potential underlying mechanisms using experimental canine models and computer simulations. METHODS: A canine model of MR was created by cauterization of mitral chordae followed by radiofrequency ablation-induced left bundle-branch block (LBBB) after 4 weeks (MR-LBBB group). Animals with LBBB alone served as control. Echocardiography was performed at baseline, acutely after MR induction, and at 4 and 20 weeks, and correlated with histology and computer simulations. RESULTS: Acute MR augmented LA reservoir and contractile strain (40±4 to 53±6% and -11±5 to -22±9% respectively, p<0.05). LA fractional area change increased significantly (47±4 to 56±4%, p<0.05) while LA end-systolic area remained unchanged (7.2±1.1 versus 7.9±1.1 cm2 respectively, p = 0.08). LA strain 'pseudonormalized' after 4 weeks and decompensated at 20 weeks with both strains decreasing to 25±6% and -3±2% respectively (p<0.05) together with a progressive increase in LA end-systolic area (7.2±1.1 to 14.0±6.3 cm2, p<0.05). In the LBBB-group, LA remodeling was less pronounced. Histology showed a trend towards increased interstitial fibrosis in the LA of the MR-LBBB group. Computer simulations indicated that the progressive changes in LA structure and function are a combination of progressive eccentric remodeling and fibrosis. CONCLUSION: MR augmented LA strain acutely to supranormal values without significant LA dilation. However, over time, LA strain gradually decreases (pseudornormal and decompensated) with LA dilation. Histology and computer simulations indicated a correlation to a varying degree of LA eccentric remodeling and fibrosis.

Uniportal video-assisted thoracoscopy is a safe approach in patients with empyema requiring surgery.

BACKGROUND: Empyema is a well-known complication of pneumonia, with high morbidity and mortality rates. This warrants direct treatment either with antibiotics and chest tube drainage or surgery. With less invasive surgical approaches such as uniportal video-assisted thoracoscopic surgery (uVATS), surgical intervention gets a more prominent role early on in the treatment of empyema. The aim of this study was to compare uVATS with the complete VATS (cVATS) approach in empyema, with respect to postoperative complications, hospital length of stay and mortality. METHODS: All cases of empyema that were treated surgically in our hospital between 2006 and 2019 were included in a retrospective database. The preferential surgical approach changed from cVATS from 2006 to 2015, towards uVATS from 2016 and on, based on the experience of the surgical team. The database included pre- and postoperative data, as well as peropartive characteristics. RESULTS: One hundred and thirty-seven patients were treated with cVATS and 49 with uVATS. Apart from a slightly reduced kidney function in the uVATS group (57.3±6.3 vs. 71.4±17.2 mL/min/1.73 m2, P≤0.001), there were no significant baseline differences in patient characteristics. The duration of uVATS was comparable to cVATS (70±17 vs. 56±23 min, P=0.240), and with low per- and postoperative complications. The postoperative hospital stay was equal in both groups (19±13 vs. 20±15 days, P=0.320). There were no statistically significant differences in postoperative complications or death. CONCLUSIONS: Uniportal VATS is a feasible and safe technique for the use in patients with empyema requiring surgery. Even if decortication in stage III empyema is required this can be performed by uniportal VATS.

Echocardiographic Assessment of Left Bundle Branch-Related Strain Dyssynchrony: A Comparison With Tagged MRI.

Recent studies have shown the efficacy of myocardial strain estimated using speckle tracking echocardiography (STE) in predicting response to cardiac resynchronisation therapy. This study focuses on circumferential strain patterns, comparing STE-acquired strains to tagged-magnetic resonance imaging (MRI-T). Second, the effect of regularisation was examined. Two-dimensional parasternal ultrasound (US) and MRI-T data were acquired in the left ventricular short-axis view of canines before (n = 8) and after (n = 9) left bunch branch block (LBBB) induction. US-based strain analysis was performed on Digital Imaging and Communications in Medicine data at the mid-level using three overall methods ("Commercial software," "Basic block-matching," "regularised block-matching"). Moreover, three regularisation approaches were implemented and compared. MRI-T analysis was performed using SinMod. Normalised regional circumferential strain curves, based on standard six or septal/lateral segments, were analysed and cross-correlated with MRI-T data. Systolic strain (SS) and septal rebound stretch (SRS) were calculated and compared. Overall agreement of normalised circumferential strain was good between all methods on a global and regional level. All STE methods showed a bias (≥4% strain) toward higher SS estimates. Pre-LBBB, septal and lateral segment correlation was excellent between the Basic (mean ρ = 0.96) and regularised (mean ρ = 0.97) methods and MRI-T. The Commercial method showed a significant discrepancy between the two walls (septal ρ = 0.94, lateral ρ = 0.68). Correlation with MRI-T reduced between pre- and post-LBBB (Commercial ρ = 0.79, Basic ρ = 0.82, mean regularised ρ = 0.86). Septal strain patterns and SRS varied with the STE software and type of regularisation, with all STE methods estimating non-zero SRS values pre-LBBB. Absolute values showed moderate agreement, with a bias for higher strain from STE. SRS varied with the type of software and extra regularisation applied. Open efforts are needed to understand the underlying causes of differences between STE methods before standardisation can be achieved. This is particularly important given the apparent clinical value of strain-based parameters such as SRS.

Local microRNA-133a downregulation is associated with hypertrophy in the dyssynchronous heart.

AIMS: Left bundle branch block (LBBB) creates considerable regional differences in mechanical load within the left ventricle (LV). We investigated expression of selected microRNAs (miRs) in relation to regional hypertrophy and fibrosis in LBBB hearts and their reversibility upon cardiac resynchronization therapy (CRT). METHODS AND RESULTS: Eighteen dogs were followed for 4 months after induction of LBBB, 10 of which received CRT after 2 months. Five additional dogs served as control. LV geometric changes were determined by echocardiography and myocardial strain by magnetic resonance imaging tagging. Expression levels of miRs, their target genes: connective tissue growth factor (CTGF), serum response factor (SRF), nuclear factor of activated T cells (NFATc4), and cardiomyocyte diameter and collagen deposition were measured in the septum and LV free wall (LVfw). In LBBB hearts, LVfw and septal systolic circumferential strain were 200% and 50% of control, respectively. This coincided with local hypertrophy in the LVfw. MiR-133a expression was reduced by 33% in the LVfw, which corresponded with a selective increase of CTGF expression in the LVfw (279% of control). By contrast, no change was observed in SRF and NFATc4 expression was decreased in LBBB hearts. CRT normalized strain patterns and reversed miR-133a and CTGF expression towards normal, expression of other miRs, related to remodelling, such as miR-199b and miR-155f, were not affected. CONCLUSIONS: In the clinically relevant large animal model of LBBB, a close inverse relation exists between local hypertrophy and miR-133a. Reduced miR-133a correlated with increased CTGF levels but not with SRF and NFATc4.

Prediction of optimal cardiac resynchronization by vectors extracted from electrograms in dyssynchronous canine hearts.

INTRODUCTION: Proper optimization of atrioventricular (AV) and interventricular (VV) intervals can improve the response to cardiac resynchronization therapy (CRT). It has been demonstrated that the area of the QRS complex (QRSarea) extracted from the vectorcardiogram can be used as a predictor of optimal CRT-device settings. We explored the possibility of extracting vectors from the electrograms (EGMs) obtained from pacing electrodes and of using these EGM-based vectors (EGMVs) to individually optimize acute hemodynamic CRT response. METHODS AND RESULTS: Biventricular pacing was performed in 13 dogs with left bundle branch block (LBBB) of which five also had myocardial infarction (MI), using 100 randomized AV- and VV-settings. Settings providing an acute increase in LV dP/dtmax ≥ 90% of the highest achieved value were defined as optimal. The prediction capability of QRSarea derived from the EGMV (EGMV-QRSarea) was compared with that of QRS duration. EGMV-QRSarea strongly correlated to the change in LV dP/dtmax (R = -0.73 ± 0.19 [LBBB] and -0.66 ± 0.14 [LBBB + MI]), while QRS duration was more poorly related to LV dP/dtmax changes (R = -0.33 ± 0.25 [LBBB] and -0.47 ± 0.39 [LBBB + MI]). This resulted in a better prediction of optimal CRT-device settings by EGMV-QRSarea than by QRS duration (LBBB: AUC = 0.89 [0.86-0.93] vs. 0.76 [0.69-0.83], P < 0.01; LBBB + MI: AUC = 0.91 [0.84-0.99] vs. 0.82 [0.59-1.00], P = 0.20, respectively). CONCLUSION: In canine hearts with chronic LBBB with or without MI, the EGMV-QRSarea predicts acute hemodynamic CRT response and identifies optimal AV and VV settings accurately. These data support the potency of EGM-based vectors as a noninvasive, easy and patient-tailored tool to optimize CRT-device settings.

Assessment of left ventricular mechanical dyssynchrony in left bundle branch block canine model: Comparison between cine and tagged MRI.

PURPOSE: To compare cine and tagged magnetic resonance imaging (MRI) for left ventricular dyssynchrony assessment in left bundle branch block (LBBB), using the time-to-peak contraction timing, and a novel approach based on cross-correlation. MATERIALS AND METHODS: We evaluated a canine model dataset (n = 10) before (pre-LBBB) and after induction of isolated LBBB (post-LBBB). Multislice short-axis tagged and cine MRI images were acquired using a 1.5 T scanner. We computed contraction time maps by cross-correlation, based on the timing of radial wall motion and of circumferential strain. Finally, we estimated dyssynchrony as the standard deviation of the contraction time over the different regions of the myocardium. RESULTS: Induction of LBBB resulted in a significant increase in dyssynchrony (cine: 13.0 ± 3.9 msec for pre-LBBB, and 26.4 ± 5.0 msec for post-LBBB, P = 0.005; tagged: 17.1 ± 5.0 msec at for pre-LBBB, and 27.9 ± 9.8 msec for post-LBBB, P = 0.007). Dyssynchrony assessed by cine and tagged MRI were in agreement (r = 0.73, P = 0.0003); differences were in the order of time difference between successive frames of 20 msec (bias: -2.9 msec; limit of agreement: 10.1 msec). Contraction time maps were derived; agreement was found in the contraction patterns derived from cine and tagged MRI (mean difference in contraction time per segment: 3.6 ± 13.7 msec). CONCLUSION: This study shows that the proposed method is able to quantify dyssynchrony after induced LBBB in an animal model. Cine-assessed dyssynchrony agreed with tagged-derived dyssynchrony, in terms of magnitude and spatial direction. J. MAGN. RESON. IMAGING 2016;44:956-963.

Ex vivo and in vivo administration of fluorescent CNA35 specifically marks cardiac fibrosis.

Cardiac fibrosis is a major hallmark of cardiac diseases. For evaluation of cardiac fibrosis, the development of highly specific and preferably noninvasive methods is desired. Our aim was to evaluate CNA35, a protein known to specifically bind to collagen, as a specific marker of cardiac fibrosis. Fluorescently labeled CNA35 was applied ex vivo on tissue sections of fibrotic rat, mouse, and canine myocardium. After quantification of CNA35, sections were examined with picrosirius red (PSR) and compared to CNA35. Furthermore, fluorescently labeled CNA35 was administered in vivo in mice. Hearts were isolated, and CNA35 labeling was examined in tissue sections. Serial sections were histologically examined with PSR. Ex vivo application of CNA35 showed specific binding to collagen and a high correlation with PSR (Pearson r  =  .86 for mice/rats and r  =  .98 for canine; both p < .001). After in vivo administration, CNA35 labeling was observed around individual cardiomyocytes, indicating its ability to penetrate cardiac endothelium. High correlation was observed between CNA35 and PSR (r  =  .91, p < .001). CNA35 specifically binds to cardiac collagen and can cross the endothelial barrier. Therefore, labeled CNA35 is useful to specifically detect collagen both ex vivo and in vivo and potentially can be converted to a noninvasive method to detect cardiac fibrosis.

A patent ductus arteriosus complicating cardiopulmonary bypass for combined coronary artery bypass grafting and aortic valve replacement only discovered by computed tomography 3D reconstruction.

We describe the case of a 59-year old male patient undergoing combined coronary artery bypass grafting and aortic valve replacement. Manipulation of the heart during cardiopulmonary bypass significantly decreased venous return. Several measures were necessary to improve venous return to a level at which continuation of the procedure was safe. Based on the initial troubles with venous return, we decided to selectively cross-clamp the aorta. This resulted in a large amount of backflow of oxygenated blood from the left ventricle, necessitating additional vents in the pulmonary artery and directly in the left ventricle. The procedure was continued uneventfully, and postoperative recovery was without significant complications. Postoperative 2D computed tomography did not show any signs of a shunt, but 3D reconstruction showed a small patent ductus arteriosus.

Electrophysiological and haemodynamic effects of vernakalant and flecainide in dyssynchronous canine hearts.

AIMS: About one-third of patients with mild dyssynchronous heart failure suffer from atrial fibrillation (AF). Drugs that convert AF to sinus rhythm may further slowdown ventricular conduction. We aimed to investigate the electrophysiological and haemodynamic effects of vernakalant and flecainide in a canine model of chronic left bundle branch block (LBBB). METHODS AND RESULTS: Left bundle branch block was induced in 12 canines. Four months later, vernakalant or flecainide was administered using a regime, designed to achieve clinically used plasma concentrations of the drugs, n = 6 for each drug. Epicardial electrical contact mapping showed that both drugs uniformly prolonged myocardial conduction time. Vernakalant increased QRS width significantly less than flecainide (17 ± 13 vs. 34 ± 15%, respectively). Nevertheless, both drugs equally decreased LVdP/dtmax by ∼15%, LVdP/dtmin by ∼10%, and left ventricular systolic blood pressure by ∼5% (P = n.s. between drugs). CONCLUSIONS: Vernakalant prolongs ventricular conduction less than flecainide, but both drugs had a similar, moderate negative effect on ventricular contractility and relaxation. Part of these reductions seems to be related to the increase in dyssynchrony.

Electrophysiological and hemodynamic effects of vernakalant and flecainide during cardiac resynchronization in dyssynchronous canine hearts.

INTRODUCTION: Patients with heart failure and left bundle branch block (LBBB) are frequently treated with biventricular pacing (BiVP). Approximately one-third of them suffer from atrial fibrillation. Pharmacological conversion of atrial fibrillation is performed with drugs that slow ventricular conduction, but the effects of these drugs on the benefit of BiVP are poorly understood. METHODS: Experiments were performed in dogs with chronic LBBB, investigating the effects of Vernakalant and Flecainide (n = 6 each) on hemodynamics and electrophysiology during epicardial (EPI) and endocardial BiVP. The degree of dyssynchrony and conduction slowing was quantified using QRS width and EPI electrical mapping. RESULTS: Compared with LBBB, EPI and endocardial BiVP reduced QRS duration by 7% ± 9% (P < 0.05 compared with LBBB) and 20% ± 13% (P < 0.05 compared with LBBB, P < 0.05 between modes), respectively. During BiVP, the administration of Vernakalant and Flecainide increased QRS duration by 20% ± 14% (P < 0.05 compared with predrug BiVP) and 34% ± 10% (P < 0.05 compared with predrug BiVP, P < 0.05 between drugs). left ventricular (LV) dP/dtmax decreased by 16% ± 8% (P < 0.05 compared with predrug BiVP) during Vernakalant and by 14% ± 15% (P < 0.05 compared with predrug BiVP) during Flecainide. The drugs did not affect the relative changes in QRS width and LV dP/dtmax induced by BiVP. CONCLUSIONS: Vernakalant and Flecainide decrease contractility, slow myocardial conduction velocity, and increase activation time. The electrical and hemodynamic benefits of BiVP are not altered by the drugs.

Acute electrical and hemodynamic effects of multisite left ventricular pacing for cardiac resynchronization therapy in the dyssynchronous canine heart.

BACKGROUND: Multisite left ventricular (multi-LV) epicardial pacing has been proposed as an alternative to conventional single-site LV (single-LV) pacing to increase the efficacy of cardiac resynchronization therapy. OBJECTIVE: To compare the effects of multi-LV versus single-LV pacing in dogs with left bundle branch block (LBBB). METHODS: Studies were performed in 9 anaesthetized dogs with chronic LBBB using 7 LV epicardial electrodes. Each electrode was tested alone and in combination with 1, 2, 3, and 6 other electrodes, the sequence of which was chosen on the basis of practical real-time electrical mapping to determine the site of the latest activation. LV total activation time (LVTAT) and dispersion of repolarization (DRep) were measured by using approximately 100 electrodes around the ventricles. LV contractility was assessed as the maximum derivative of left ventricular pressure (LVdP/dtmax ). RESULTS: Single-LV pacing provided, on average, a -4.0% ± 9.3% change in LVTAT and 0.2% ± 13.7% change in DRep. Multi-LV pacing markedly decreased both LVTAT and DRep in a stepwise fashion to reach -41.3% ± 5% (P < .001 for overall comparison) and -14.2% ± 19.5% (P < .02 for overall comparison) in the septuple-LV pacing configuration, respectively. Single-LV pacing provided a mean increase of 10.7% ± 7.7% in LVdP/dtmax. LVdP/dtmax incrementally increased by the addition of pacing electrodes to 16.4% ± 8.7% (P < .001 for overall comparison). High response to single-LV pacing could not be improved further during multi-LV pacing. CONCLUSIONS: Compared with single-LV pacing, multi-LV pacing can considerably reduce both LVTAT and DRep in dogs with LBBB, but the improvement in contractility is limited to conditions where single-LV pacing provides suboptimal improvement. Further studies are warranted to determine whether these acute effects translate in antiarrhythmic properties and better long-term outcomes.

Interplay of electrical wavefronts as determinant of the response to cardiac resynchronization therapy in dyssynchronous canine hearts.

BACKGROUND: The relative contribution of electromechanical synchronization and ventricular filling to the optimal hemodynamic effect in cardiac resynchronization therapy (CRT) during adjustment of stimulation-timings is incompletely understood. We investigated whether optimal hemodynamic effect in CRT requires collision of pacing-induced and intrinsic activation waves and optimal filling of the left ventricle (LV). METHODS AND RESULTS: CRT was performed in dogs with chronic left bundle-branch block (n=8) or atrioventricular (AV) block (n=6) through atrial (A), right ventricular (RV) apex, and LV-basolateral pacing. A 100 randomized combinations of A-LV/A-RV intervals were tested. Total activation time (TAT) was calculated from >100 contact mapping electrodes. Mechanical interventricular dyssynchrony was determined as the time delay between upslopes of LV and RV pressure curves. Settings providing an increase in LVdP/dtmax (maximal rate of rise of left ventricular pressure) of ≥90% of the maximum LVdP/dtmax value were defined as optimal (CRTopt). Filling was assessed by changes in LV end-diastolic volume (EDV; conductance catheter technique). In all hearts, CRTopt was observed during multiple settings, providing an average LVdP/dtmax increase of ≈15%. In AV-block hearts, CRTopt exclusively depended on interventricular-interval and not on AV-interval. In left bundle-branch block hearts, CRTopt occurred at A-LV intervals that allowed fusion of LV-pacing-derived activation with right bundle-derived activation. In all animals, CRTopt occurred at settings resulting in the largest decrease in TAT and mechanical interventricular dyssynchrony, whereas LV EDV hardly changed. CONCLUSIONS: In left bundle-branch block and AV-block hearts, optimal hemodynamic effect of CRT depends on optimal interplay between pacing-induced and intrinsic activation waves and the corresponding mechanical resynchronization rather than filling.

Transseptal conduction as an important determinant for cardiac resynchronization therapy, as revealed by extensive electrical mapping in the dyssynchronous canine heart.

BACKGROUND: Simple conceptual ideas about cardiac resynchronization therapy assume that biventricular (BiV) pacing results in collision of right and left ventricular (LV) pacing-derived wavefronts. However, this concept is contradicted by the minor reduction in QRS duration usually observed. We investigated the electric mechanisms of cardiac resynchronization therapy by performing detailed electric mapping during extensive pacing protocols in dyssynchronous canine hearts. METHODS AND RESULTS: Studies were performed in anesthetized dogs with acute left bundle-branch block (LBBB, n=10) and chronic LBBB with tachypacing-induced heart failure (LBBB+HF, n=6). Activation times (AT) were measured using LV endocardial contact and noncontact mapping and epicardial contact mapping. BiV pacing reduced QRS duration by 21±10% in LBBB but only by 5±12% in LBBB+HF hearts. Transseptal impulse conduction was significantly slower in LBBB+HF than in LBBB hearts (67±9 versus 44±16 ms, respectively), and in both groups significantly slower than transmural LV conduction (≈30 ms). In both groups QRS duration and vector and the epicardial AT vector amplitude and angle were significantly different between LV and BiV pacing, whereas the endocardial AT vector was similar. During variation of atrioventricular delay while LV pacing, and ventriculo-ventricular delay while BiV pacing, the optimal hemodynamic effect was achieved when epicardial AT and QRS vectors were minimal and endocardial AT vector indicated LV preexcitation. CONCLUSIONS: Due to slow transseptal conduction, the LV electric activation sequence is similar in LV and BiV pacing, especially in failing hearts. Optimal hemodynamic cardiac resynchronization therapy response coincides with minimal epicardial asynchrony and QRS vector and LV preexcitation.

Animal models of dyssynchrony.

Cardiac resynchronization therapy (CRT) is an important therapy for patients with heart failure and conduction pathology, but the benefits are heterogeneous between patients and approximately a third of patients do not show signs of clinical or echocardiographic response. This calls for a better understanding of the underlying conduction disease and resynchronization. In this review, we discuss to what extent established and novel animal models can help to better understand the pathophysiology of dyssynchrony and the benefits of CRT.