Human Left Atrium Research Articles

Comparison of LGE-MRI and Local Impedance Data Recorded in Human Left Atria

Abstract Efficient personalized ablation strategies for treating atrial arrhythmias remain challenging. Discrepancies in identifying arrhythmogenic areas using characterization methods, such as late gadolinium enhanced magnetic resonance imaging (LGE-MRI) and electroanatomical mapping, require a comparative analysis of local impedance (LI) and LGE-MRI data. This study aims to analyze correlations as basis for improvement of treatment strategies. 16 patients undergoing left atrium (LA) ablation with LGE-MRI acquisition and LI data recording were recruited. LGE-MRI data and LI measurements were normalized to patient- and modality-specific blood pool references. A global mean shape was generated based on all patient geometries, and normalized local impedance (LIN) and LGE-MRI image intensity ratio (IIR) data points were coregistered for comparison. Data analysis comprised intra-patient and inter-patient assessments, evaluating differences in LIN values among datasets categorized by their IIR. Due to substantial deviations in LIN values, even within the same patient and IIR-category, discerning the presence or absence of a correlation was challenging, and no statistically significant correlation could be identified. Our findings underscore the necessity for standardized protocols in data acquisition, processing, and comparison, to minimize unquantified confounding effects. While immediate substitution of LI for LGE-MRI seems improbable given the significant LIN variations, this preliminary study lays the groundwork for systematic data acquisition. By ensuring data quality, a meaningful comparison between LI and LGE-MRI data can be facilitated, potentially shaping future strategies for atrial arrhythmia treatment.

Transcriptomic Insights into the Atrial Fibrillation Susceptibility Locus near the MYOZ1 and SYNPO2L Genes

Genome-wide association studies have identified a locus on chromosome 10q22, where many co-inherited single nucleotide polymorphisms (SNPs) are associated with atrial fibrillation (AF). This study seeks to identify the impact of this locus on gene expression at the transcript isoform level in human left atria and to gain insight into potential causal variants. Bulk RNA sequencing was analyzed to identify myozenin 1 (MYOZ1) and synaptopodin 2-like (SYNPO2L) transcript isoforms and the association of common SNPs in this region with transcript isoform expression levels. Chromatin marks were used to suggest candidate regulatory SNPs in this region. Protein amino acid changes were examined for predicted functional consequences. Transfection of MYOZ1 and two SYNPO2L isoforms were performed to localize their encoded proteins in cardiomyocytes derived from stem cells. We identified one MYOZ1 transcript isoform and four SYNPO2L transcript isoforms, two of which encode proteins, while the other two encode long noncoding RNAs (lncRNAs). The risk allele of the strongest AF susceptibility SNP on chromosome 10q22 is associated with decreased MYOZ1 expression and increased expression of the two SNYPO2L lncRNA isoforms. There are many SNPs co-inherited with the top AF-associated SNP due to linkage disequilibrium (LD), including rs11000728, which we propose as the MYOZ1 regulatory SNP, confirmed by reporter gene transfection. In addition, this LD block includes three missense SNPs in the SYNPO2L gene, with the minor protective haplotype predicted to be detrimental to protein function. MYOZ1 and both protein isoforms of SYNPO2L were localized to the sarcomere. This is a complex locus with the potential for several SNPs in a haplotype to alter AF susceptibility by opposing effects on MYOZ1 and SYNPO2L lncRNA expression, along with effects on SYNPO2L protein function.

PITX2 deficiency leads to atrial mitochondrial dysfunction.

Reduced left atrial PITX2 is associated with atrial cardiomyopathy and atrial fibrillation. PITX2 is restricted to left atrial cardiomyocytes in the adult heart. The links between PITX2 deficiency, atrial cardiomyopathy and atrial fibrillation are not fully understood. To identify mechanisms linking PITX2 deficiency to atrial fibrillation, we generated and characterized PITX2-deficient human atrial cardiomyocytes derived from human induced pluripotent stem cells (hiPSC) and their controls. PITX2-deficient hiPSC-derived atrial cardiomyocytes showed shorter and disorganised sarcomeres and increased mononucleation. Electron microscopy found an increased number of smaller mitochondria compared to the control. Mitochondrial protein expression was altered in PITX2-deficient hiPSC-derived atrial cardiomyocytes. Single-nuclear RNA-sequencing found differences in cellular respiration pathways and differentially expressed mitochondrial and ion channel genes in PITX2-deficient hiPSC-derived atrial cardiomyocytes. PITX2 repression in hiPSC-derived atrial cardiomyocytes replicated dysregulation of cellular respiration. Mitochondrial respiration was shifted to increased glycolysis in PITX2-deficient hiPSC-derived atrial cardiomyocytes. PITX2-deficient human hiPSC-derived atrial cardiomyocytes showed higher spontaneous beating rates. Action potential duration was more variable with an overall prolongation of early repolarization, consistent with metabolic defects. Gene expression analyses confirmed changes in mitochondrial genes in left atria from 42 patients with atrial fibrillation compared to 43 patients in sinus rhythm. Dysregulation of left atrial mitochondrial (COX7C) and metabolic (FOXO1) genes was associated with PITX2 expression in human left atria. In summary, PITX2 deficiency causes mitochondrial dysfunction and a metabolic shift to glycolysis in human atrial cardiomyocytes. PITX2-dependent metabolic changes can contribute to the structural and functional defects found in PITX2-deficient atria.

Abstract 19018: Unraveling the Regulatory Landscape of NKX2-5 in Human Left Atrial Tissue: Implications for Atrial Fibrillation

Cardiac transcription factors are emerging as crucial modulators of atrial fibrillation (AF), with rare and common variation implicated in AF. Mutations in NKX2-5 and more recently, NFATC1 were reported in families with autosomal dominant young-onset AF. Here, we explore the landscape of NKX2-5 and NFATC1 in human left atrium, using Chromatin Immunoprecipitation Sequencing (ChIP-seq), DAVID functional annotation and HOMER motif analyses. Post-mortem human left atrial tissue samples (n=5) were obtained from the National Disease Research Interchange (NDRI) within a recovery interval of < 4 hours. Preserved microstructural integrity and normal staining patters for TNNT2 and vimentin were verified using confocal microscopy. ChIP-seq analysis revealed an average of 2.62k NKX2-5 binding peaks, with 118 genes bound in all samples. Functional analysis identified significant enrichment in gene ontology categories and pathways (Table 1), including adrenergic and PPAR signaling pathways known to participate in AF susceptibility. Using HOMER, we mapped 55 NFATC1 consensus binding sites to the NKX2-5 ChIP peaks. Two NFATC1 consensus sites localized to an NKX2-5 binding peak located upstream from MEF2A (Figure 1). Published HI-C data from human right atrium indicates the NFATC1 binding sites are adjacent to the promoter of MEF2A in 3D space, suggesting that NKX2-5 and NFATC1 co-regulate MEF2A. In murine atria, mef2A regulates genes involved in fibrosis and adhesion, processes that are crucial in the pathogenesis of atrial fibrillation. Our data, together with published literature, provide a framework for understanding transcriptional networks that may predispose to human AF.

Abstract 15845: Unraveling Postoperative Atrial Fibrillation at the Single-Cell Level

Background: Postoperative atrial fibrillation (poAF) is one of the most common complications after cardiac surgery. It increases the length of stay in the ICU and hospital, increases the risk of stroke, and leads to death. Several clinical factors, including age, obesity, and co-occurring conditions such as prior atrial fibrillation, hypertension, kidney disease, etc., are associated with poAF. We hypothesize that the atrial transcriptome differs between patients who remain in sinus rhythm following surgery and those who develop poAF, allowing the identification of genes, pathways, and cell types necessary for the understanding and treatment of poAF. Methods: Single-cell RNA sequencing (scRNA-seq) was performed to gain insights into the heterogeneous cell populations of the left atrium. It provided an in-depth view of the variability in the cellular response to poAF through the molecular changes within cells, allowing for identifying biomarkers and therapeutic targets. Changes in gene expression associated with poAF were modeled, including batch, sex, age, atrial size, and history of atrial fibrillation and covariates. Results: sc-RNA-seq data of thirty-one human left atrium samples were analyzed. The samples included 14 females and 17 males aged 35-86 years. Cell clusters were identified after the quality check, preprocessing, and normalization. Nine distinct cell types were identified, including atrial cardiomyocytes, fibroblasts, endothelial cells, smooth muscle cells, adipocytes, pericytes, myeloid, lymphoid, and neuronal cells. The atrial cardiomyocytes and fibroblasts were the most abundant identified cell types. The enrichment of these cell types and significant changes in many marker genes in the disease condition suggest their crucial role in their response to the disease. Conclusion: The comprehensive analysis revealed distinct gene expression patterns and cellular heterogeneity. It provided valuable insights into the cellular landscape and transcriptional alterations in poAF. By shedding light on the molecular mechanisms, the findings pave the way for developing targeted interventions to mitigate the impact of poAF.

Novel functional atrial fibrillation risk genes and pathways identified from coexpression analyses in human left atria

Genomewide association studies have associated >100 genetic loci with atrial fibrillation (AF), but establishing causal genes contributing to AF remains challenging. The purpose of this study was to determine candidate novel causal genes and mechanistic pathways associated with AF risk loci by incorporating gene expression and coexpression analyses and to provide a resource for functional studies and targeting of AF-associated genes. Cis-expression quantitative trait loci were identified for candidate genes near AF risk variants in human left atrial tissues. Coexpression partners were identified for each candidate gene. Weighted gene coexpression network analysis (WGCNA) identified modules and modules with overrepresentation of candidate AF genes. Ingenuity pathway analysis (IPA) was applied to the coexpression partners of each candidate gene. IPA and gene set over representation analysis were applied to each WGCNA module. One hundred sixty-six AF-risk single nucleotide polymorphisms were located in 135 loci. Eighty-one novel genes not previously annotated as putative AF risk genes were identified. IPA identified mitochondrial dysfunction, oxidative stress, epithelial adherens junction signaling, and sirtuin signaling as the most frequent significant pathways. WGCNA characterized 64 modules (candidate AF genes overrepresented in 8), represented by cell injury, death, stress, developmental, metabolic/mitochondrial, transcription/translation, and immune activation/inflammation regulatory pathways. Candidate gene coexpression analyses suggest significant roles for cellular stress and remodeling in AF, supporting a dual risk model for AF: Genetic susceptibility to AF may not manifest until later in life, when cellular stressors overwhelm adaptive responses. These analyses also provide a novel resource to guide functional studies on potential causal AF genes.

THE VARIANT ANATOMY OF PULMONARY VEINS OF PEOPLE OF DIFFERENT BODY TYPES

Cardiovascular diseases occupy a central place in the structure of morbidity and mortality in various population groups and in most cases occur with a heart rhythm disorder. One of the most common types of arrhythmia is atrial fibrillation, the morphological substrate for the development of which is the left atrium and the region of the pulmonary veins. In connection with the introduction of modern imaging systems into the practice of cardiac surgery, which enable a detailed study of the anatomy of the heart and its separate chambers, there is a need to study the features of the variant anatomy of the pulmonary veins of the human left atrium in the aspect of modern interventional cardiac surgery, which will make it possible to secure and accelerate surgical interventions performed in this area. The purpose of the study was the revealing of variant anatomy of the pulmonary veins using multislice computed tomography, depending on body type and gender. Heart tomograms of 70 patients were analyzed (8 of the 1st period of adulthood, 23 of the 2nd period of adulthood, 37 of the elderly, 2 of the senile age). Among them were 47 men - 67.1% and 23 women - 32.9%. As a result of the analysis of the obtained data, four variants of the confluence of the pulmonary veins into the left atrium were identified. Among all observations, a typical variant of the confluence of the pulmonary veins into the left atrium occurs in 78.6% of cases, in 85.7% of cases - in persons of the pyknic type, in 85.0% of cases - in the examined asthenic type and in 69.0% of cases - in representatives of the normosthenic body type. Less common are the common pulmonary vein collector on the left (12.9%), as well as the accessory pulmonary vein on the right (7.1%). An extremely rare anatomical variant was the common pulmonary vein collector on both sides (1.4%). Thus, the anatomy of the distal pulmonary veins is characterized by significant variability, there is a slight relationship with gender and body type. An in-depth study of the anatomy of the pulmonary veins remains relevant for the effective implementation of surgical interventions in this anatomical zone.

A computational modeling framework for pre-clinical evaluation of cardiac mapping systems.

There are a variety of difficulties in evaluating clinical cardiac mapping systems, most notably the inability to record the transmembrane potential throughout the entire heart during patient procedures which prevents the comparison to a relevant "gold standard". Cardiac mapping systems are comprised of hardware and software elements including sophisticated mathematical algorithms, both of which continue to undergo rapid innovation. The purpose of this study is to develop a computational modeling framework to evaluate the performance of cardiac mapping systems. The framework enables rigorous evaluation of a mapping system's ability to localize and characterize (i.e., focal or reentrant) arrhythmogenic sources in the heart. The main component of our tool is a library of computer simulations of various dynamic patterns throughout the entire heart in which the type and location of the arrhythmogenic sources are known. Our framework allows for performance evaluation for various electrode configurations, heart geometries, arrhythmias, and electrogram noise levels and involves blind comparison of mapping systems against a "silver standard" comprised of computer simulations in which the precise transmembrane potential patterns throughout the heart are known. A feasibility study was performed using simulations of patterns in the human left atria and three hypothetical virtual catheter electrode arrays. Activation times (AcT) and patterns (AcP) were computed for three virtual electrode arrays: two basket arrays with good and poor contact and one high-resolution grid with uniform spacing. The average root mean squared difference of AcTs of electrograms and those of the nearest endocardial action potential was less than 1ms and therefore appears to be a poor performance metric. In an effort to standardize performance evaluation of mapping systems a novel performance metric is introduced based on the number of AcPs identified correctly and those considered spurious as well as misclassifications of arrhythmia type; spatial and temporal localization accuracy of correctly identified patterns was also quantified. This approach provides a rigorous quantitative analysis of cardiac mapping system performance. Proof of concept of this computational evaluation framework suggests that it could help safeguard that mapping systems perform as expected as well as provide estimates of system accuracy.

Secretome of atrial epicardial adipose tissue facilitates reentrant arrhythmias by myocardial remodeling.

Epicardial adipose tissue (EAT) accumulation is associated with cardiac arrhythmias. The effect of EAT secretome (EATs) on cardiac electrophysiology remains largely unknown. The purpose of this study was to investigate the arrhythmogenicity of EATs and its underlying molecular and electrophysiological mechanisms. We collected atrial EAT and subcutaneous adipose tissue (SAT) from 30 patients with atrial fibrillation (AF), and EAT from 3 donors without AF. The secretome was collected after a 24-hour incubation of the adipose tissue explants. We cultured neonatal rat ventricular myocytes (NRVMs) with EATs, subcutaneous adipose tissue secretome (SATs), and cardiomyocytes conditioned medium (CCM) for 72 hours. We implemented the electrophysiological changes observed after EATs incubation into a model of human left atrium and tested arrhythmia inducibility. Incubation of NRVMs with EATs decreased expression of the potassium channel subunit Kcnj2 by 26% and correspondingly reduced the inward rectifier K+ current IK1 by 35% compared to incubation with CCM, resulting in a depolarized resting membrane of cardiomyocytes. EATs decreased expression of connexin43 (29% mRNA, 46% protein) in comparison to CCM. Cells incubated with SATs showed no significant differences in Kcnj2 or Gja1 expression in comparison to CCM, and their resting potential was not depolarized. Cardiomyocytes incubated with EATs showed reduced conduction velocity and increased conduction heterogeneity compared to SATs and CCM. Computer modeling of human left atrium revealed that the electrophysiological changes induced by EATs promote sustainedreentrant arrhythmias if EAT partially covers the myocardium. EAT slows conduction, depolarizes the resting potential, alters electrical cell-cell coupling, and facilitates reentrant arrhythmias.

Cardiac endocardial left atrial substrate and lesion depth mapping using near-infrared spectroscopy.

Atrial fibrillation (AF) is a rapid irregular electrical activity in the upper chamber and the most common sustained cardiac arrhythmia. Many patients require radiofrequency ablation (RFA) therapy to restore sinus rhythm. Pulmonary vein isolation requires distinguishing normal atrial wall from the pulmonary vein tissue, and atrial substrate ablation requires differentiating scar tissue, fibrosis, and adipose tissue. However, current anatomical mapping methods for strategically locating ablation sites by identifying structural substrates in real-time are limited. An intraoperative tool that accurately provides detailed structural information and classifies endocardial substrates could help improve RF guidance during RF ablation therapy. In this work, we propose a 7F NIRS integrated ablation catheter and demonstrate endocardial mapping on ex vivo swine (n = 12) and human (n = 5) left atrium (LA). First, pulmonary vein (PV) sleeve, fibrosis and ablation lesions were identified with NIRS-derived contrast indices. Based on these key spectral features, classification algorithms identified endocardial substrates with high accuracy (<11% error). Then, a predictive model for lesion depth was evaluated on classified lesions. Model predictions correlated well with histological measurements of lesion dimensions (R = 0.984). Classified endocardial substrates and lesion depth were represented in 2D spatial maps. These results suggest NIRS integrated mapping catheters can serve as a complementary tool to the current electroanatomical mapping system to improve treatment efficacy.

Cardiac chamber-specific genetic alterations suggest candidate genes and pathways implicating the left ventricle in the pathogenesis of atrial fibrillation.

It is believed that the atria play a predominant role in the initiation and maintenance of atrial fibrillation (AF), while the role of left ventricular dysfunction in the pathophysiology remains enigmatic. We sought to dissect chamber specificity of AF-associated transcriptional changes using RNA-sequencing. We performed intra- and inter-chamber differential expression analyses comparing AF against sinus rhythm to identify genes specifically dysregulated in human left atria, right atria, and left ventricle (LV), and integrated known AF genetic associations with expression quantitative trait loci datasets to inform the potential for disease causal contributions within each chamber. Inter-chamber patterns changed drastically. Vast AF-associated transcriptional changes specific to LV, enriched for biological pathway terms implicating mitochondrial function, developmental processes and immunity, were supported at the genetic level, but no major enrichments for candidate genes specific to the atria were found. Our observations suggest an active role of the LV in the pathogenesis of AF.

P984Proof of endo-epicardial asynchrony of the left atrial wall in humans

Abstract Funding Acknowledgements Prof. Dr. NMS de Groot is supported by funding grants from CVON-AFFIP (914728), NWO-Vidi (91717339), Biosense Webster USA (ICD 783454) and Medical Del Introduction Treatment of atrial fibrillation (AF) is still suboptimal as mechanisms underlying AF initiation and persistence are incompletely understood. Endo-Epicardial asynchrony (EEA) plays an important role in AF persistence and has so far only been demonstrated in the right atrium (RA). Purpose To investigate whether EEA also exists in the thin walled left atrium (LA) and to measure the maximal degree of EEA between the endo- and epicardial layers during sinus rhythm (SR). Methods Simultaneous endo-epicardial mapping of the LA was performed during SR in 3 male patients (73 ± 1.5 years) with history of paroxysmal AF undergoing cardiac surgery including rhythm surgery and LA appendage amputation. Simultaneous endo-epicardial mapping was performed with a mapping clamp containing two electrode arrays of 8x16 electrodes (diameters: 0.4mm, interelectrode distance: 2mm) positioned exactly opposite to each other. The mapping clamp was introduced through the LA appendage with its tip towards the superior pulmonary vein. Local endo-epicardial activation time differences were determined by selecting the median time delay within the exact opposite electrode and its 8 surrounding electrodes. The asynchrony map consisted of the maximum of 2 medians from direct opposite electrodes. EEA was defined as time differences ≥15ms. Conduction delay (CD) and conduction block (CB) were defined as differences in local activation times between neighboring electrodes of respectively ≥7 and ≥12ms. Results A total of 35 SR beats were analyzed. Mean total activation time of the whole endo-epicardial LA tissue was 42.4 ± 9.5ms and did not differ between both layers (epicardium: 31.2 ± 9.9ms; endocardium: 37.8 ± 10.3ms; P= 0.62). CD and CB were observed in respectively 3.2% and 6.3% at the epicardium and 3.3% and 3.0% at the endocardium. The lowest amount of CD (5.2%) and CB (0.3%) was observed in the patient who had his first AF episode only 11 days prior to surgery. Also, no EEA was present in this patient. In two patients with paroxysmal AF &amp;gt;6 months, the prevalence of EEA was respectively 2.7% and 41.4% and the degree of EEA ranged from 15 to 44ms. Interestingly, the patient with the highest degree of EEA was diagnosed with paroxysmal AF for almost 5 years (Figure 1). Conclusion Our data provides evidence for the existence of EEA in the human left atrium which appears to be already present during SR. Knowledge of EEA and the ability to stage AF based on the degree of EEA is essential for individualized and staged future therapy for AF. Abstract Figure 1. The maximal degree of endo-epi

Fast Quasi-Conformal Regional Flattening of the Left Atrium.

Two-dimensional representation of 3D anatomical structures is a simple and intuitive way for analysing patient information across populations and image modalities. While cardiac ventricles, especially the left ventricle, have an established standard representation (bull's eye plot), the 2D depiction of the left atrium (LA) remains challenging due to its sub-structural complexity including the pulmonary veins (PV) and the left atrial appendage (LAA). Quasi-conformal flattening techniques, successfully applied to cardiac ventricles, require additional constraints in the case of the LA to place the PV and LAA in the same geometrical 2D location for different cases. Some registration-based methods have been proposed but surface registration is time-consuming and prone to errors when the geometries are very different. We propose a novel atrial flattening methodology where a 2D standardised map of the LA is obtained quickly and without errors related to registration. The LA is divided into five regions which are then mapped to their analogue two-dimensional regions. 67 human left atria from magnetic resonance images (MRI) were studied to derive a population-based template representing the averaged relative locations of the PVs and LAA. The clinical application of our methodology is illustrated on different use cases including the integration of MRI and electroanatomical data.

Validation of Dipole Density Mapping During Atrial Fibrillation and SinusRhythm in Human Left Atrium.

This study sought to validate the accuracy of noncontact electrograms against contact electrograms in the left atrium during sinus rhythm (SR) and atrial fibrillation (AF). Noncontact mapping offers the opportunity to assess global cardiac activation in the chamber of interest. A novel noncontact mapping system, which records intracardiac voltage to derive cellular charge sources (dipoledensity), allows real-time mapping of AF to guide ablation. Noncontact and contact unipolar electrogram pairs were recorded simultaneously from multiple locations. Morphology correlation and timing difference of reconstructed electrograms obtained from a noncontact catheter were compared with those from contact electrograms obtained from a contact catheter at the same endocardial locations. A total of 796 electrogram pairs in SR and 969 electrogram pairs in AF were compared from 20 patients with persistent AF. The median morphology correlation and timing difference (ms) in SR was 0.85 (interquartile range [IQR]: 0.71 to 0.94) and 6.4ms (IQR: 2.6 to 17.1 ms); in AF was 0.79 (IQR: 0.69 to 0.88) and 14.4ms (IQR: 6.7 to 26.2 ms), respectively. The correlation was stronger and the timing difference was less when the radial distance (r) from the noncontact catheter center to the endocardium was≤ 40 versus > 40mm; 0.87(IQR: 0.72 to 0.94) versus 0.73 (IQR: 0.56 to 0.88) and 5.7ms (IQR:2.6 to 15.4 ms) versus 15.1ms (IQR: 4.1 to 27.7 ms); p<0.01 when in SR; 0.81(IQR: 0.69 to 0.89) versus 0.67(IQR: 0.45 to 0.82) and 12.3ms (IQR: 5.9 to 21.8 ms) versus 28.3ms (IQR: 16.2 to 36.0 ms); p<0.01 when in AF. This novel noncontact dipole density mapping system provides comparable reconstructed atrialelectrogrammeasurements in SR or AF in human left atrium when the anatomical site of interest is≤40mm fromthe mapping catheter.

Imaging of subendocardial adipose tissue and fiber orientation distributions in the human left atrium using optical coherence tomography.

BackgroundOptical coherence tomography (OCT) has the potential to provide real‐time imaging guidance for atrial fibrillation ablation, with promising results for lesion monitoring. OCT can also offer high‐resolution imaging of tissue composition, but there is insufficient cardiac OCT data to inform the use of OCT to reveal important tissue architecture of the human left atrium. Thus, the objective of this study was to define OCT imaging data throughout the human left atrium, focusing on the distribution of adipose tissue and fiber orientation as seen from the endocardium.Methods and ResultsHuman hearts (n = 7) were acquired for imaging the left atrium with OCT. A spectral‐domain OCT system with 1325 nm center wavelength, 6.5 μm axial resolution, 15 μm lateral resolution, and a maximum imaging depth of 2.51 mm in the air was used. Large‐scale OCT image maps of human left atrial tissue were developed, with adipose thickness and fiber orientation extracted from the imaging data. OCT imaging showed scattered distributions of adipose tissue around the septal and pulmonary vein regions, up to a depth of about 0.43 mm from the endocardial surface. The total volume of adipose tissue detected by OCT over one left atrium ranged from 1.42 to 28.74 mm3. Limited fiber orientation information primarily around the pulmonary veins and the septum could be identified.ConclusionOCT imaging could provide adjunctive information on the distribution of subendocardial adipose tissue, particularly around thin areas around the pulmonary veins and septal regions. Variations in OCT‐detected tissue composition could potentially assist ablation guidance.

Abstract 656: Construction and Application of an Epigenetic Atlas of the Human Heart

Epigenetic modifications, including DNA methylation, regulate gene expression and contribute to the differentiation of cells. Tissues are characterized by methylation patterns that reflect their specific functions and origin. Currently, there is limited knowledge of the epigenetic patterns of the heart. To address this gap, we generated methylation profiles for all anatomical regions of the human heart. We hypothesize that unique differentially methylated regions (DMRs) of DNA exist for each cardiac region and that these patterns are disrupted in heart failure. Using non-diseased cardiac tissue and reduced representation bisulfite sequencing on a Illumina platform, we generated genome-wide methylomes for the human right atrium (n=4), left atrium (n=4), right ventricle (n=4), left ventricle (n=4), aorta (n=3), pulmonary artery (n=2), mitral valve (n=3), tricuspid valve (n=3), aortic valve (n=3) and pulmonary valve (n=3). DMRs, defined as regions with significantly different mean methylation differences, were identified using Metilene. For each tissue we identified between 10-20 million reads covering 8-10 million CpG methylation sites and 4-228 tissue-specific DMRs. There were relatively few (4) different DMRs between the left and right ventricles but 228 unique DMRs were found in the vessels (aorta and pulmonary artery) compared to the ventricles including regions upstream of BMP3 and FOXC1 , genes implicated in cardiogenesis. We then applied this approach and normal data to the analysis of disease. We generated additional left ventricular methylomes from adult (n=3) and pediatric (n=3) patients with heart failure. We identified 19 unique DMRs in the adult group and 107 in the pediatric group. Genes associated with pediatric HF DMRs included ROCK1 and FBLN2 that have been previously implicated in contraction and remodelling. We have created an atlas of the human heart based upon differences in DNA methylation between the anatomical regions of the human heart. This data will help increase our understanding of cardiac development, identify new disease biomarkers and have application to a wide range of cardiac diseases

Age-related regulation and region-specific distribution of ion channel subunits promoting atrial fibrillation in human left and right atria.

Age-induced changes and electrical remodelling are important components of the atrial fibrillation (AF) substrate. To study regional distribution and age-dependent changes in gene expression that may promote AF in human atria. Human left atrial (LA) and right atrial (RA) tissue samples were obtained from donor hearts unsuitable for transplantation and from patients undergoing mitral valve repair. Atrial fibrillation was mimicked in vitro by tachypacing of human atrial tissue slices. Ionic currents were studied by the whole-cell patch-clamp technique; gene expression was analysed by real-time qPCR and immunoblotting. Both healthy RA and RA from older patients showed greater CACNA1c mRNA and CaV1.2 protein expression than LA. No age-dependent changes of Kir2.1 expression in both atria were seen. Remodelling occurred in a qualitatively similar manner in RA and LA. IK1 and Kir2.1 protein expression increased with AF. MiR-1, miR-26a, and miR-26b were down-regulated with AF in both atria. ICa,L was decreased. CACNA1c and CACNA2b expression decreased and miR-328 increased in RA and LA during AF. Ex vivo tachypacing of human atrial slices replicated these findings. There were age-dependent increases in miR-1 and miR-328, while miR-26a decreased with age in atrial tissues from healthy human donor hearts. Features of electrical remodelling in man occur in a qualitatively similar manner in both human atria. Age-related miR-328 dysregulation and reduced ICa,L may contribute to increased AF susceptibility with age.

Early Tissue Reaction After Second-Generation Cryoballoon Ablation Evaluated with Intracardiac Echocardiography.

Radiofrequency energy applications immediately produce tissue edema. This study aimed to investigate the acute tissue reaction immediately after second-generation cryoballoon applications using 3-dimensional intracardiac echocardiography (ICE) imaging technology.This study consisted of 10 patients with paroxysmal atrial fibrillation who underwent pulmonary vein isolation (PVI) using second-generation cryoballoons. Ablation was performed with a single 3-minute freeze strategy and exclusively 28-mm balloons. The left atrial and right pulmonary vein (PV) antra geometries were created with 3-dimensional ICE technology before and immediately after the PVI.Out of 20 right PVs, 19 were isolated exclusively with cryoballoons, and one right inferior PV (RIPV) required touch-up ablation. All 10 right superior PVs (RSPVs) were isolated by single cryoballoon applications, and RIPVs were isolated by a mean of 1.2 ± 0.4 applications. The total application time was 171 ± 19 and 203 ± 71 seconds, and nadir balloon temperature was -56.0 ± 4.9 and -53.8 ± 5.4°C for the RSPVs and RIPVs, respectively. In all patients, diffuse wall thickening of the antra and ostium of the right PVs was observed as compared to baseline. The wall thickening was 0-0.25 mm in 3 patients, and 0.25-0.5 mm in the remaining 7. During the median follow-up of 13 [10.2-17.2] months, 8 (80%) patients were free from arrhythmia recurrences. Nine (90%) patients underwent repeat cardiac computed tomography at a median of 6.0 [4.5-12.0] months after the initial procedure, and no PV stenosis was observed in all.Tissue edema and wall thickening appeared in the human left atrium immediately after second-generation cryoballoon ablation.

Feasibility of percutaneous epicardial mapping and ablation for refractory atrial fibrillation: Insights into substrate and lesion transmurality

Recurrences of atrial fibrillation (AF) after ablation have been attributed to conduction gaps and nontransmural ablation lesions. The purpose of this study was to assess the feasibility of adjunctive percutaneous mapping of the epicardial regions of the left atrium to characterize the transmural extent of substrate and ablation lesions. Between 2014 and 2018, combined epicardial and endocardial mapping of AF was performed in 18 patients via an inferior subxiphoid percutaneous approach (16 with previously failed ablation procedures and 2 patients with long-standing persistent AF) at 2 centers. Epicardial substrate mapping was compared with endocardial mapping to assess transmural uniformity. Of 18 patients, 4 (22%) demonstrated nontransmural atrial low-voltage regions with relative epicardial sparing in the left atrial posterior wall. Transmural isolation of the posterior wall was achieved after an endocardial "box" lesion set in 6/9 (67%), guided by epicardial voltage data, while epicardial and endocardial dissociation during AF was observed in 1 patient. In 3 patients, epicardial capture along the endocardial pulmonary vein lesion set despite endocardial capture loss and bidirectional block was observed. Two cases of mitral flutter were terminated from the epicardium. A balloon was positioned in the pericardial space in 6 patients for esophageal protection during ablation. A percutaneous epicardial approach for mapping and ablation of the left atrium is feasible in the electrophysiology laboratory during endocardial catheter ablation for AF and may be useful as an adjunctive approach in refractory cases. High-density epicardial mapping can provide direct evidence of nonuniform lesion and substrate transmurality of the human left atrium before and after ablation.

Structural remodeling and conduction velocity dynamics in the human left atrium: Relationship with reentrant mechanisms sustaining atrial fibrillation

BackgroundRate-dependent conduction velocity (CV) slowing is associated with atrial fibrillation (AF) initiation and reentrant mechanisms.ObjectiveThe purpose of this study was to assess the relationship between bipolar voltage, CV dynamics, and AF drivers.MethodsPatients undergoing catheter ablation for persistent AF (<24 months) were enrolled. Unipolar electrograms were recorded with a 64-pole basket catheter during atrial pacing at 4 pacing intervals (PIs) during sinus rhythm. CVs were measured between pole pairs along the wavefront path and correlated with underlying bipolar voltage. CV dynamics within low-voltage zones (LVZs <0.5 mV) were compared to those of non-LVZs (≥0.5 mV) and were correlated to driver sites mapped using CARTOFINDER (Biosense Webster).ResultsEighteen patients were included (age 62 ± 10 years). Mean CV at 600 ms was 1.59 ± 0.13 m/s in non-LVZs vs 0.98 ± 0.23 m/s in LVZs (P <.001). CV decreased incrementally over all 4 PIs in LVZs, whereas in non-LVZs a substantial decrease in CV was only seen between PIs 300–250 ms (0.59 ± 0.09 m/s; P <.001). Rate-dependent CV slowing sites measurements, defined as exhibiting CV reduction ≥20% more than the mean CV reduction seen between PIs 600–250 ms for that voltage zone, were predominantly in LVZs (0.2–0.5 mV; 75.6% ± 15.5%; P <.001). Confirmed rotational drivers were mapped to these sites in 94.1% of cases (sensitivity 94.1%, 95% CI 71.3%–99.9%; specificity 77.9%, 95% CI 74.9%–80.7%).ConclusionCV dynamics are determined largely by the extent of remodeling. Rate-dependent CV slowing sites are predominantly confined to LVZs (0.2–0.5 mV), and the resultant CV heterogeneity may promote driver formation in AF.